Tumor targeting protein or its fragment, antibody binding thereto and use thereof

Abstract

The present invention relates to a tumor targeting protein or a fragment thereof, an antibody bound thereto and its use. More specifically, the present invention relates to a vector comprising a nucleic acid encoding protein A56 or a fragment thereof and its use. In addition, the present invention relates to an antibody binding to protein A56 or its fragment and its use. The present invention uses an oncolytic virus as a vector, and the vector contains a nucleic acid encoding protein A56 of the present invention, or a fragment thereof, or a mutant thereof, so when applied to an individual, it mainly specifically kills only cancer cells. In addition, even cancer cells that survive after being infected with an oncolytic virus express protein A56 on the cell surface, which enables it to be targeted for secondary anticancer treatment. Therefore, according to one embodiment of the present invention, when a vector containing a nucleic acid encoding protein A56 or a fragment thereof, and an antibody binding to A56 is used, cancer can be effectively treated.

Description

Technical Field

[0001] The present invention relates to a tumor targeting protein or a fragment thereof, an antibody binding thereto and uses thereof. More specifically, the present invention relates to a vector comprising a nucleic acid encoding a protein A56 fragment and uses thereof. In addition, the present invention relates to an antibody specifically binding to protein A56 or a fragment thereof and uses thereof. Background Art

[0002] Cancer, also known as tumor, refers to cells that proliferate abnormally due to the body's own excessive growth. Due to the aging population, the increase in smoking, the increase in alcohol consumption, the westernization of eating habits and environmental pollution in modern society, the incidence of cancer continues to rise.

[0003] Methods for treating cancer include surgery, radiotherapy, chemotherapy, and others. Specifically, surgery is a treatment method that removes cancerous tissue from the body, and is very effective for early-stage cancer or cancer that is confined to a specific location. However, cancer that has invaded the tissues surrounding the lesion or has metastasized to the lymph nodes is difficult to remove, and the possibility of such cancer recurring is high. In such cases, radiotherapy or chemotherapy is used in combination with surgery. Radiotherapy or chemotherapy is mainly used to treat advanced or terminal cancer; however, this treatment can also affect normal cells, resulting in serious side effects.

[0004] Recently, immune cell therapy has been actively studied as a cancer treatment method. Immunocell therapy differs from existing treatments in that it uses the patient's immune cells to kill cancer cells. Specifically, immune cell therapy is a method of obtaining immune cells from the patient's body, activating them to specifically attack proliferating cells or cancer cells, and then returning them to the patient's body. This method maximizes the anti-cancer effect while minimizing the side effects caused by the drug.

[0005] In addition, over the past 5 years, immune checkpoint inhibitors have been actively studied for use in immunotherapy to treat cancer. In particular, inhibitors targeting immune checkpoints, such as cytotoxic T lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD-1), and PD-L1 have been studied. Immune checkpoint inhibitors, such as ipilimumab (anti-CTLA-4), nivolumab (anti-PD-1), and pembrolizumab (anti-PD-1), have been approved by regulatory agencies for the treatment of multiple types of cancer. However, immune checkpoint inhibitors are limited to the treatment of certain types of cancer, such as melanoma, lung cancer, head and neck cancer, kidney cancer, and bladder cancer.

[0006] Therefore, there is a need for continued research and development of cancer cell targeting approaches that are safe and can even effectively target solid cancers. Summary of the invention

[0007] Technical issues

[0008] Therefore, in order to develop a method for safely and effectively targeting cancer cells, the present inventors conducted research and found that when cancer cells are treated by injecting a vector containing a nucleic acid encoding protein A56 or a fragment thereof, protein A56 is expressed on the surface of cancer cells. In addition, the present inventors found that antibodies that bind to protein A56 or a fragment thereof bind to protein A56 expressed on the surface of cancer cells, thereby completing the present invention.

[0009] Solutions to technical problems

[0010] In one aspect of the present invention, an anticancer agent is provided, which comprises an antibody or a fragment thereof that specifically binds to protein A56 or a fragment thereof as an active ingredient.

[0011] In another aspect of the present invention, an oncolytic virus is provided, which comprises a nucleic acid encoding protein A56 or a fragment thereof.

[0012] In another aspect of the present invention, an antibody or a fragment thereof that binds to protein A56 or a fragment thereof is provided.

[0013] In another aspect of the present invention, a cancer-targeting polypeptide or a fragment thereof is provided, wherein the polypeptide comprises an amino acid sequence shown by SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079.

[0014] In another aspect of the present invention, an isolated nucleic acid encoding the cancer-targeting polypeptide or a fragment thereof is provided.

[0015] In another aspect of the present invention, a kit for treating cancer is provided, comprising the oncolytic virus and the antibody or a fragment thereof.

[0016] In another aspect of the present invention, a pharmaceutical composition for targeting cancer is provided, comprising the oncolytic virus.

[0017] Advantageous Effects of the Invention

[0018] The oncolytic virus of the present invention contains nucleic acid encoding protein A56 or its fragment or variant, and when applied to an individual as a vector, it mainly kills only cancer cells in a specific manner. In addition, even if the cancer cells infected with the oncolytic virus are still alive, they express protein A56 on the cell surface, which enables targeted secondary anticancer therapy. Therefore, in the case of using a vector comprising a nucleic acid encoding protein A56 or its fragment or variant, and an antibody binding protein A56 or its fragment, cancer can be effectively treated, each of which is an embodiment of the present invention.

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 A diagram showing the process of targeting tumor cells using oncolytic viruses containing nucleic acids encoding protein A56 or fragments thereof.

[0021] Figure 2 Shown are the results obtained by infecting a human lung cancer cell line (A549) with an oncolytic virus containing a nucleic acid encoding protein A56, and then identifying whether protein A56 is expressed on the cell surface of the infected A549 cancer cell line using immunofluorescence staining.

[0022] Figure 3 Shown are the results obtained by infecting a human colorectal cancer cell line (HCT-116) with an oncolytic virus containing a nucleic acid encoding protein A56, and then identifying whether protein A56 is expressed on the cell surface of the infected HCT-116 cell line using immunofluorescence staining.

[0023] Figure 4 Shown are the results obtained by intraperitoneally administering an oncolytic virus containing a nucleic acid encoding protein A56 to mice transplanted with a human colorectal cancer cell line (HT-29), and then identifying the expression of protein A56 on the surface of various tissues.

[0024] Figure 5 Shown are the results obtained by intraperitoneally administering an oncolytic virus containing a nucleic acid encoding protein A56 to normal rabbits, and then identifying the expression of protein A56 on the surface of each tissue.

[0025] Figure 6 Shown are the results obtained by intratumorally administering an oncolytic virus containing a nucleic acid encoding protein A56 to mice transplanted with a mouse renal carcinoma cell line (Renca), followed by identifying the expression of protein A56 on the surface of tumor tissue within 7, 10 and 14 days.

[0026] Figure 7The results obtained by intratumorally administering an oncolytic virus containing a nucleic acid encoding protein A56 and hydroxyurea to mice transplanted with a mouse renal cancer cell line (Renca), and then identifying the expression of protein A56 on the surface of tumor tissue on days 7, 10, and 14 are shown.

[0027] Figure 8 Shown are the results obtained by administering an oncolytic virus containing a nucleic acid encoding protein A56 twice to mice transplanted with a mouse renal cancer cell line (Renca), and then identifying the expression of protein A56 on the surface of tumor tissue on days 21, 24, and 28.

[0028] Fig. 9 The results obtained by secondary administration of oncolytic virus containing nucleic acid encoding protein A56 and hydroxyurea to mice transplanted with a mouse renal cancer cell line (Renca), and then identifying the expression of protein A56 on the surface of tumor tissue on days 21, 24 and 28 are shown.

[0029] Fig.10 Shown are the results obtained by administering an oncolytic virus containing a nucleic acid encoding protein A56 and hydroxyurea to mice transplanted with a mouse renal cancer cell line (Renca), and then measuring the tumor volume of the mice.

[0030] Fig.11 Shown are the results obtained by administering an oncolytic virus containing a nucleic acid encoding protein A56 and hydroxyurea to mice transplanted with a mouse renal cancer cell line (Renca), and then measuring the body weight of the mice.

[0031] Fig.12 A schematic diagram showing an embodiment of protein A56 and fragments thereof is shown.

[0032] Fig.13 Shown are the results obtained by identifying the expression of protein A56 and its fragments on the cell surface.

[0033] Figures 14 to 19 Shown are the results obtained by measuring the affinity between the prepared anti-A56 antibodies and protein A56.

[0034] FIG. 20A shows the results of measuring the production amounts of antibodies A56-01A02 to A56-02B06.

[0035] FIG. 20B shows a graph of the production of antibodies A56-01A02 to A56-02B06.

[0036] FIG. 20C shows the results of measuring the production amounts of antibodies A56-02D04 to A56-59E12.

[0037] Figures 21 to 37Shown are the results obtained by purifying the produced anti-A56 antibodies and then identifying the antibodies by SDS-PAGE.

[0038] Fig.38 Shown are the results of HeLa or HT-29 cell lines that had been treated or not with oncolytic viruses, treated with each of the five anti-A56 antibodies, and then measured by flow cytometry.

[0039] Fig.39 Shown are the results of A549, MCF-7, or PC-3 cell lines that had been treated or not with oncolytic viruses, treated with each of the three anti-A56 antibodies, and then measured by flow cytometry.

[0040] Fig.40 Shown are the results of HeLa cell lines that had been treated or not treated with oncolytic viruses, treated with anti-A56 antibody (Antibodies-Online), and then subjected to FACS analysis.

[0041] Fig.41 Shown are the results of HeLa cell lines that had been treated or not treated with oncolytic viruses, treated with anti-A56 antibody (LakePharma variant 3), and then subjected to FACS analysis.

[0042] Fig.42 Shown are the results of HeLa cell lines that had been treated or not treated with oncolytic viruses, treated with anti-A56 antibody (LakePharma variant 4), and then subjected to FACS analysis.

[0043] Fig.43 Shown are the results of HeLa cell lines that had been treated or not treated with oncolytic viruses, treated with anti-A56 antibodies (Antibody Online, LakePharma Variant 3, or LakePharma Variant 4), and then subjected to FACS analysis.

[0044] Fig.44 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab18), and then measured by flow cytometry.

[0045] Fig.45 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab19), and then measured by flow cytometry.

[0046] Fig.46 Shown are the results obtained by treating A549 cancer cell lines that were or were not treated with oncolytic viruses (OTS-412, WOTS-418), with anti-A56 antibody (AblO1), and then measuring by flow cytometry.

[0047] Fig.47 Shown are the results obtained by treating A549 cancer cell lines that were or were not treated with oncolytic viruses (OTS-412, WOTS-418), with anti-A56 antibody (Ab13), and then measuring by flow cytometry.

[0048] Fig.48 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab14), and then measured by flow cytometry.

[0049] Fig.49 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab08), and then measured by flow cytometry.

[0050] Fig.50 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab03), and then measured by flow cytometry.

[0051] Fig.51 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab51), and then measured by flow cytometry.

[0052] Fig.52 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab55), and then measured by flow cytometry.

[0053] Fig.53 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab16), and then measured by flow cytometry.

[0054] Fig.54Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 3), and then measured by flow cytometry.

[0055] Fig.55 Shown are the results of A549 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 4), and then measured by flow cytometry.

[0056] Fig.56 Shown are the results of A549 cancer cell lines treated or not with oncolytic virus (OTS-412), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0057] Fig.57 Shown are the results of A549 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0058] Fig.58 Shown are the results of A549 cancer cell lines treated or not with oncolytic virus (OTS-412), treated with anti-A56 antibodies (Ab18, Ab16), and then measured by flow cytometry.

[0059] Fig.59 Shown are the results of A549 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Abl8, Abl6), and then measured by flow cytometry.

[0060] Fig.60 Shown are the results obtained for A549 cancer cell lines that were or were not treated with an oncolytic virus (OTS-412), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0061] Fig.61 Shown are the results obtained for A549 cancer cell lines that were or were not treated with an oncolytic virus (WOTS-418), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0062] Fig.62 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab18), and then measured by flow cytometry.

[0063] Fig.63 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab19), and then measured by flow cytometry.

[0064] Fig.64 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab01), and then measured by flow cytometry.

[0065] Fig.65 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab13), and then measured by flow cytometry.

[0066] Fig.66 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab14), and then measured by flow cytometry.

[0067] Fig.67 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab08), and then measured by flow cytometry.

[0068] Fig.68 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab03), and then measured by flow cytometry.

[0069] Fig.69 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab51), and then measured by flow cytometry.

[0070] Fig.70Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab55), and then measured by flow cytometry.

[0071] Fig.71 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab16), and then measured by flow cytometry.

[0072] Fig.72 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 3), and then measured by flow cytometry.

[0073] Fig.73 Shown are the results of PC3 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 4), and then measured by flow cytometry.

[0074] Fig.74 Shown are the results of PC3 cancer cell lines treated or not with oncolytic virus (OTS-412), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0075] Fig.75 Shown are the results of PC3 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0076] Fig.76 Shown are the results of PC3 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Abl8, Abl6), and then measured by flow cytometry.

[0077] Fig.77 Shown are the results of PC3 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Abl8, Abl6), and then measured by flow cytometry.

[0078] Fig.78Shown are the results obtained from PC3 cancer cell lines that were or were not treated with an oncolytic virus (OTS-412), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0079] Fig.79 Shown are the results obtained from PC3 cancer cell lines that were or were not treated with an oncolytic virus (WOTS-418), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0080] Fig.80 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab18), and then measured by flow cytometry.

[0081] Fig.81 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab19), and then measured by flow cytometry.

[0082] Fig.82 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab01), and then measured by flow cytometry.

[0083] Fig.83 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab13), and then measured by flow cytometry.

[0084] Fig.84 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab14), and then measured by flow cytometry.

[0085] Fig.85 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab08), and then measured by flow cytometry.

[0086] Fig.86Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab03), and then measured by flow cytometry.

[0087] Fig.87 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab51), and then measured by flow cytometry.

[0088] Fig.88 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab55), and then measured by flow cytometry.

[0089] Fig.89 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (Ab16), and then measured by flow cytometry.

[0090] Fig.90 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 3), and then measured by flow cytometry.

[0091] Fig.91 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic viruses (OTS-412, WOTS-418), treated with anti-A56 antibody (LP variant 4), and then measured by flow cytometry.

[0092] Fig.92 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic virus (OTS-412), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0093] Fig.93 Shown are the results of MCF7 cancer cell lines treated or not with oncolytic virus (WOTS-418), treated with anti-A56 antibodies (Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16), and then measured by flow cytometry.

[0094] Fig.94Shown are the results obtained for MCF7 cancer cell lines that were or were not treated with an oncolytic virus (OTS-412), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0095] Fig.95 Shown are the results obtained for MCF7 cancer cell lines that were or were not treated with an oncolytic virus (WOTS-418), treated with LakePharma variant 3 or LakePharma variant 4, and then measured by flow cytometry.

[0096] Fig.96 The results obtained by performing ELISA using the prepared antibodies are shown.

[0097] Fig.97 Shown are the results obtained by identifying the expression level of Fc-fused A56 fusion protein.

[0098] Fig.98 Shown are the results obtained by identifying the expression levels of the prepared A56 fragments.

[0099] Fig.99 Results obtained by analyzing sequence homology with respect to protein A56 in oncolytic viruses (OTS-412 and WOTS-418) are shown.

[0100] Best Mode for Carrying Out the Invention

[0101] Hereinafter, the present invention will be described in detail.

[0102] In one aspect of the present invention, an anticancer agent is provided, which comprises an antibody or a fragment thereof that specifically binds to protein A56 or a fragment thereof as an active ingredient.

[0103] As used herein, the term "protein A56" refers to a protein translated from a gene represented by A56, A56R or HA (e.g., Gene ID: 3707652), which is encoded in the gene of a Poxviridae virus when a host cell is infected with a Poxviridae virus. Protein A56 or a fragment thereof may include an amino acid sequence shown by SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079. The nucleic acid encoding protein A56 or a fragment thereof may include the nucleotide sequence shown by SEQ ID NO:1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1072, 1074, 1076 or 1078.

[0104] Specifically, protein A56 may be wild-type protein A56 or a variant thereof. Wild-type protein A56 may have an amino acid sequence shown by SEQ ID NO: 1038 or 1073. In addition, the nucleic acid encoding wild-type protein A56 may be a nucleotide sequence shown by SEQ ID NO: 1039 or 1072.

[0105] In addition, the protein A56 variant may be substituted, deleted or added with one or more amino acids, as long as the variant can be located on the surface of cancer cells like protein A56. The nucleotide sequence encoding the protein A56 variant may be a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO: 1038 or 1073 having at least 60%, at least 70%, at least 80% or at least 90% sequence homology, and may most preferably be a nucleotide sequence encoding an amino acid sequence having at least 95% sequence homology thereto. Specifically, the protein A56 variant may include the amino acid sequence shown by SEQ ID NO: 1075 or 1079. The nucleic acid encoding the protein A56 variant may include the nucleotide sequence shown by SEQ ID NO: 1074 or 1078.

[0106] The protein A56 fragment may be a polypeptide comprising the amino acid sequence shown by SEQ ID NO: 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060 or 1077. In addition, the nucleotide sequence encoding the fragment may be a nucleotide sequence encoding the polypeptide shown by SEQ ID NO: 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060 or 1076. Specifically, the nucleotide sequence encoding the polypeptide shown by SEQ ID NO:1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060 or 1077 can be represented by SEQ ID NO:1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061 or 1076 in the above order.

[0107] In addition, the nucleotide sequence encoding the protein A56 fragment can be a nucleotide sequence encoding an amino acid sequence having at least 60%, at least 70%, at least 80% or at least 90% sequence homology with the amino acid sequence shown in SEQ ID NO:1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060 or 1077, and most preferably a nucleotide sequence encoding an amino acid sequence having at least 95% sequence homology thereto.

[0108] As used herein, the term "antibody" refers to an immune protein that binds to an antigen and interferes with its action or removes the antigen. There are five types of antibodies: IgM, IgD, IgG, IgA and IgE, each containing a heavy chain produced by a heavy chain constant region gene μ, δ, γ, α or ε. In antibody technology, IgG is mainly used. IgG includes four isotypes: IgG1, IgG2, IgG3 and IgG4, each of which may have different structural and functional properties.

[0109] IgG forms a very stable Y-shaped structure (molecular weight: about 150 kDa) composed of two heavy chain (about 50 kDa) proteins and two light chain (about 25 kDa) proteins. Antibodies have light chains and heavy chains, and each chain is divided into a variable region with different amino acid sequences between antibodies and a constant region with the same amino acid sequence between antibodies. The heavy chain constant region includes CH1, H (hinge), CH2, and CH3 domains. Each domain is composed of two β-folds, which are connected by intramolecular disulfide bonds. The two variable regions in the heavy and light chains combine to form an antigen binding site. This site is present in the two arms of the Y shape. In the Y shape, the part that can bind to the antigen is called the antibody binding fragment (Fab), and the part that does not bind to the antigen is called the crystallizable fragment (Fc). Fab and Fc are connected by a flexible hinge region.

[0110] As used herein, the term "CDR" refers to the site that binds to the antigen, which is a hypervariable region present in the variable region of the heavy and light chains of the antibody, and its amino acid sequence varies from antibody to antibody. From the three-dimensional structure of the antibody, the CDR is ring-shaped on the surface of the antibody; below the ring, there is a framework region (FR) that structurally supports the CDR. Each heavy chain and light chain has three ring structures, and these six ring region structures are combined with each other to directly contact the antigen. For convenience, the antigen binding sites on the six ring region structures are respectively referred to as CDR1, CDR2, CDR3, CDR4, CDR5 and CDR6.

[0111] In addition, the antibody fragment may be any one selected from the group consisting of Fab, scFv, F(ab)2 and Fv.Antibody fragment refers to the antigen binding domain, excluding the crystallizable region (Fc region) having effector functions of transmitting antigen binding stimulation to cells, complement, etc., and may include third generation antibody fragments such as single domain antibodies or mini antibodies.

[0112] In addition, the antibody fragment has the following advantages: the antibody fragment is smaller than the fully structured IgG, thereby increasing the permeability to tissues or tumors; and the antibody fragment can be produced in bacteria, reducing production costs. In addition, the antibody fragment does not have Fc, so it is used in cases where it is not desired to transmit antigen binding stimulation to cells, complement, etc. The antibody fragment has a short half-life in the human body and is therefore suitable for in vivo diagnosis; however, replacing some of the basic, acidic or neutral amino acids that constitute the antibody may change the unique isoelectric point (pI) of the antibody. This change in the isoelectric point of the antibody can cause changes including, for example, reducing the in vivo toxic side effects of the antibody or increasing the water solubility of the antibody. Therefore, for therapeutic antibodies, considering affinity or structural form, a fully structured IgG can be used.

[0113] The antibody can be easily produced by known monoclonal antibody production technology. The method for producing monoclonal antibodies can be carried out by preparing hybridomas using B lymphocytes obtained from immune animals, or can be carried out by using phage display technology. However, the present invention is not limited to this.

[0114] The antibody or fragment thereof may include:

[0115] A heavy chain variable region (VH) comprising a heavy chain CDR1 selected from the group consisting of SEQ ID NO: 1, 18, 35, 52, 69, 86, 103, 120, 137, 154, 171, 188, 205, 222, 239, 256, 273, 290, 307, 324, 341, 358, 375, 392, 409, 426, 443, 460, 477, 494, 511, 528, 54 5, 562, 579, 596, 613, 630, 647, 664, 681, 698, 715, 732, 749, 766, 783, 800, 817, 834, 851, 868, 885, 902, 919, 936, 953, 970, 987, 1004, 1023, 1062 and 1063;

[0116] A heavy chain CDR2 selected from the group consisting of SEQ ID NO: 2, 19, 36, 53, 70, 87, 104, 121, 138, 155, 172, 189, 206, 223, 240, 257, 274, 291, 308, 325, 342, 359, 376, 393, 410, 427, 444, 461, 478, 495, 512, 529, 546, 563, 580, 597, 614, 631, 648, 665, 682, 699, 716, 733, 750, 767, 784, 80 1, 818, 835, 852, 869, 886, 903, 920, 937, 954, 971, 988, 1005, 1022, and 1064; and

[0117] A heavy chain CDR3 selected from the group consisting of SEQ ID NO: 3, 20, 37, 54, 71, 88, 105, 122, 139, 156, 173, 190, 207, 224, 241, 258, 275, 292, 309, 326, 343, 360, 377, 394, 411, 428, 445, 462, 479, 496, 513, 530, 547, 564, 581, 598, 615, 632, 649, 666, 683, 700, 717, 734, 751, 768, 785, 80 2, 819, 836, 853, 870, 887, 904, 921, 938, 955, 972, 989, 1006, 1023 and 1065; and

[0118] A light chain variable region (VL), comprising a light chain CDR1 selected from the group consisting of: SEQ ID NO:4, 21, 38, 55, 72, 89, 106, 123, 140, 157, 174, 191, 208, 225, 242, 259, 2 76, 293, 310, 327, 344, 361, 378, 395, 412, 429, 446, 463, 480, 497, 514, 531 ,548,565,582,599,616,633,650,667,684,701,718,735,752,769,786,8 03, 820, 837, 854, 871, 888, 905, 922, 939, 956, 973, 990, 1007, 1024 and 1066;

[0119] A light chain CDR2 selected from the group consisting of SEQ ID NO: 5, 22, 39, 56, 73, 90, 107, 124, 141, 158, 175, 192, 209, 226, 243, 260, 277, 294, 311, 328, 345, 362, 379, 396, 413, 430, 447, 464, 481, 498, 515, 532, 549, 566, 583, 600, 617, 634, 651, 668, 685, 702, 719, 736, 753, 770, 787, 80 4, 821, 838, 855, 872, 889, 906, 923, 940, 957, 974, 991, 1008, 1025 and 1067; and

[0120] Light chain CDR3 selected from the group consisting of: SEQ ID NO: 6, 23, 40, 57, 74, 91, 108, 125, 142, 159, 176, 193, 210, 227, 244, 261, 2 78, 295, 312, 329, 346, 363, 380, 397, 414, 431, 448, 465, 482, 499, 516, 533 ,550,567,584,601,618,635,652,669,686,703,720,737,754,771,788,8 05, 822, 839, 856, 873, 890, 907, 924, 941, 958, 975, 992, 1009, 1026 and 1068.

[0121] The antibody or fragment thereof may include:

[0122] a heavy chain variable region (VH) comprising a heavy chain CDR1 having an amino acid sequence as shown in SEQ ID NO: 1, 35, 120, 222, 239, 290, 324, 341, 851, 919, 1062 or 1063;

[0123] A heavy chain CDR 2 having the amino acid sequence shown in SEQ ID NO: 2, 36, 121, 223, 240, 291, 325, 342, 852, 920 or 1064; and

[0124] A heavy chain CDR3 having the amino acid sequence shown in SEQ ID NO: 3, 37, 122, 224, 241, 292, 326, 343, 853, 921 or 1065; and

[0125] a light chain variable region (VL) comprising a light chain CDR1 having an amino acid sequence as shown in SEQ ID NO: 4, 38, 123, 225, 242, 293, 327, 344, 854, 922 or 1066;

[0126] A light chain CDR2 having the amino acid sequence shown in SEQ ID NO: 5, 39, 124, 226, 243, 294, 328, 345, 855, 923 or 1067; and

[0127] A light chain CDR3 having the amino acid sequence shown in SEQ ID NO:6, 40, 125, 227, 244, 295, 329, 346, 856, 924 or 1068.

[0128] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO:1; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO:2; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO:3; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO:4; a light chain CDR2 having an amino acid sequence of SEQ ID NO:5; and a light chain CDR3 having an amino acid sequence of SEQ ID NO:6.

[0129] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO:35; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO:36; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO:37; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO:38; a light chain CDR2 having an amino acid sequence of SEQ ID NO:39; and a light chain CDR3 having an amino acid sequence of SEQ ID NO:40.

[0130] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 120; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 121; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 122; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 123; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 124; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 125.

[0131] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 222; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 223; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 224; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 225; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 226; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 227.

[0132] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 239; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 240; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 241; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 242; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 243; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 244.

[0133] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 290; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 291; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 292; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 293; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 294; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 295.

[0134] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 324; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 325; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 326; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 327; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 328; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 329.

[0135] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 341; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 342; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 343; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 344; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 345; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 346.

[0136] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO:851; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO:852; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO:853; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO:854; a light chain CDR2 having an amino acid sequence of SEQ ID NO:855; and a light chain CDR3 having an amino acid sequence of SEQ ID NO:856.

[0137] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO:919; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO:920; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO:921; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO:922; a light chain CDR2 having an amino acid sequence of SEQ ID NO:923; and a light chain CDR3 having an amino acid sequence of SEQ ID NO:924.

[0138] The antibody or fragment thereof may include a heavy chain variable region (VH), which includes a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 1062 or 1063; a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 1064; and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 1065; and a light chain variable region (VL), which includes a light chain CDR1 having an amino acid sequence of SEQ ID NO: 1066; a light chain CDR2 having an amino acid sequence of SEQ ID NO: 1067; and a light chain CDR3 having an amino acid sequence of SEQ ID NO: 1068.

[0139] In one embodiment, the antibody or fragment thereof may include the following CDR combinations:

[0140] [Table 1]

[0141]

[0142]

[0143] In particular, the antibody or fragment thereof may include a heavy chain variable region having an amino acid sequence selected from the group consisting of SEQ ID NO: 8, 25, 42, 59, 76, 93, 110, 127, 144, 161, 178, 195, 212, 229, 246, 263, 280, 297, 314, 331, 348, 365, 382, ​​399, 416, 433, 450, 467, 484, 501, 518, 535, 55 2, 569, 586, 603, 620, 637, 654, 671, 688, 705, 722, 739, 756, 773, 790, 807, 824, 841, 858, 875, 892, 909, 926, 943, 960, 977, 994, 1011, 1028, 1069 and 1070.

[0144] Specifically, the antibody or fragment thereof may include a heavy chain variable region having any one amino acid sequence selected from the following group: SEQ ID NO: 8, 42, 127, 229, 246, 297, 331, 348, 858, 926, 1069 and 1070.

[0145] In addition, the antibody or fragment thereof may include a light chain variable region having any one amino acid sequence selected from the group consisting of: SEQ ID NO:9, 26, 43, 60, 77, 94, 111, 128, 145, 162, 179, 196, 213, 230, 247, 264, 2 81, 298, 315, 332, 349, 366, 383, 400, 417, 434, 451, 468, 485, 502, 519, 536 ,553,570,587,604,621,638,655,672,689,706,723,740,757,774,791,8 08, 825, 842, 859, 876, 893, 910, 927, 944, 961, 978, 995, 1012, 1029 and 1071.

[0146] Specifically, the antibody or fragment thereof may include a light chain variable region having any one amino acid sequence selected from the following group: SEQ ID NO: 9, 43, 128, 230, 247, 298, 332, 349, 859, 927, 1071.

[0147] The antibody or fragment thereof may include a heavy chain variable region having any one of the amino acid sequences selected from the group consisting of SEQ ID NO: 8, 25, 42, 59, 76, 93, 110, 127, 144, 161, 178, 195, 212, 229, 246, 263, 280, 297, 314, 331, 348, 365, 382, ​​399, 416, 433, 450, 467, 484, 501, 518, 535, 552, 569, 586, 603 , 620, 637, 654, 671, 688, 705, 722, 739, 756, 773, 790, 807, 824, 841, 858, 875, 892, 909, 926, 943, 960, 977, 994, 1011, 1028, 1069 and 1070; and a light chain variable region of any one amino acid sequence selected from the group consisting of: SEQ ID NO:9, 26, 43, 60, 77, 94, 111, 128, 145, 162, 179, 196, 213, 230, 247, 264, 2 81, 298, 315, 332, 349, 366, 383, 400, 417, 434, 451, 468, 485, 502, 519, 536 ,553,570,587,604,621,638,655,672,689,706,723,740,757,774,791,8 08, 825, 842, 859, 876, 893, 910, 927, 944, 961, 978, 995, 1012, 1029 and 1071.

[0148] Specifically, the antibody or its fragment may include a heavy chain variable region having any one amino acid sequence selected from the following group: SEQ ID NO: 8, 42, 127, 229, 246, 297, 331, 348, 858, 926, 1069 and 1070; and a light chain variable region having any one amino acid sequence selected from the following group: SEQ ID NO: 9, 43, 128, 230, 247, 298, 332, 349, 859, 927 and 1071.

[0149] Anticancer agents can target cancer cells infected with oncolytic viruses. Specifically, the oncolytic virus can be an oncolytic vaccinia virus. The oncolytic virus can contain a nucleic acid encoding protein A56 or a fragment thereof.

[0150] Protein A56 or its fragment is as described above. Protein A56 may be a wild-type protein A56 or a variant of protein A56. The nucleic acid encoding A56 protein or its fragment may be a nucleic acid encoding a wild-type A56 protein or a variant of A56 protein.

[0151] The nucleic acid encoding protein A56 or a fragment thereof may include the nucleotide sequence shown by SEQ ID NO:1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1072, 1074, 1076 or 1078.

[0152] The vaccinia virus can be, but is not limited to, one of the following vaccinia virus strains: Western Reserve (WR), New York vaccinia virus (NYVAC), Wyeth (NYCBOH), LC16m8, Lister, Copenhagen, Tian Tan, USSR, TashKent, Evans, International Health Division-J (IHD-J), and International Health Division-White (IHD-W).

[0153] An oncolytic virus is a virus in which the thymidine kinase (TK) gene is deleted. Specifically, the oncolytic virus can be a recombinant vaccinia virus in which the thymidine kinase gene is deleted.

[0154] As used herein, the term "thymidine kinase (TK)" refers to an enzyme called thymidine kinase and involved in nucleotide biosynthesis. TK is an enzyme used for nucleotide biosynthesis in cells and viruses. Here, for cells, normal cells no longer divide, so TK is not present therein; even for rapidly dividing cells, such as hair follicle cells, the amount of TK present is not enough for the virus to utilize. From these viewpoints, the deletion of the TK gene in the virus allows the virus to proliferate only in the presence of cancer cells where TK is present, thereby selectively killing cancer cells.

[0155] The anticancer agent may also include a physiologically acceptable carrier. In addition, the anticancer agent may also include suitable excipients and diluents commonly used to prepare pharmaceutical compositions. In addition, the anticancer agent may be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories, or injections according to conventional methods. Specifically, the anticancer agent may be in the form of an injection. Suitable preparations known in the art may be those disclosed in the literature (Remington Pharmaceutical Science, 1985).

[0156] Anticancer agent can be used for preventing or treating the cancer selected from the group consisting of lung cancer, colorectal cancer, prostate cancer, thyroid cancer, breast cancer, brain cancer, head and neck cancer, esophageal cancer, skin cancer, thymus cancer, stomach cancer, colon cancer, liver cancer, ovarian cancer, uterine cancer, bladder cancer, rectal cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer, lymphoma, acute leukemia, multiple myeloma or a combination thereof. In addition, for anticancer agent, the examples of carrier, excipient and diluent can include sodium chloride, lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil etc. In the case of formulating the anticancer agent, it can be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants and surfactants.

[0157] Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. For non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used. As a base for suppositories, Witepsol, polyethylene glycol, Tween 61, cocoa butter, laurel fat, glycerin gelatin, etc. may be used.

[0158] The cancer may be a solid cancer or a blood cancer. Specifically, the solid cancer may be any one selected from the group consisting of lung cancer, colorectal cancer, prostate cancer, thyroid cancer, breast cancer, brain cancer, head and neck cancer, esophageal cancer, skin cancer, thymus cancer, stomach cancer, colon cancer, liver cancer, ovarian cancer, uterine cancer, bladder cancer, rectal cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer, or a combination thereof. In addition, the blood cancer may be any one selected from the group consisting of lymphoma, acute leukemia, multiple myeloma, or a combination thereof.

[0159] The dosage of the anticancer agent can be preferably determined according to the patient's age, sex, symptoms, the amount of active ingredient absorbed in the body, the inactivation rate, drugs used in combination, etc. The anticancer agent can be administered at 0.0001 mg / kg (body weight) to 100 mg / kg (body weight) based on the antibody or its fragment.

[0160] In another aspect of the present invention, an oncolytic virus is provided, which comprises a nucleic acid encoding protein A56 or a fragment thereof.

[0161] In another aspect of the present invention, a composition for targeting cancer cells is provided, comprising an oncolytic virus as an active ingredient, wherein the oncolytic virus contains a nucleic acid encoding protein A56 or a fragment thereof. The oncolytic virus is as described in the above-mentioned anticancer agent.

[0162] Cancer cells can be cells of solid cancer or blood cancer. Specifically, the solid cancer can be, but is not limited to, any one selected from the group consisting of lung cancer, colorectal cancer, prostate cancer, thyroid cancer, breast cancer, brain cancer, head and neck cancer, esophageal cancer, skin cancer, thymus cancer, stomach cancer, colon cancer, liver cancer, ovarian cancer, uterine cancer, bladder cancer, rectal cancer, gallbladder cancer, biliary tract cancer, pancreatic cancer, kidney cancer, fibrosarcoma, testicular cancer, brain metastases, liver metastases, or a combination thereof.

[0163] Furthermore, the blood cancer may be, but is not limited to, any one selected from the group consisting of lymphoma, acute leukemia, multiple myeloma, or a combination thereof.

[0164] In yet another aspect of the present invention, a method for targeting cancer cells is provided, comprising the step of administering an oncolytic virus to a subject.

[0165] The subject can be a mammal, including a human, and can be a non-human animal. The term "non-human animal" refers to any vertebrate and can include mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians and reptiles. In addition, the subject refers to an individual suffering from a cancer disease, or the disease can be alleviated, inhibited or treated by administering an oncolytic virus.

[0166] The dose of oncolytic virus varies according to the individual's condition and body weight, disease severity, drug type, administration route and administration time, and can be appropriately selected by a person skilled in the art. The dose of oncolytic virus received by the patient is 1×10 5 to 1×10 18 Virus particles, infectious viral units (TCID 50 ) or plaque forming units (pfu). Specifically, patients received an oncolytic virus dose of 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 5×10 10 , 1×10 11 , 5×10 11 , 1×10 12, 1×10 13 , 1×10 14 , 1×10 15 , 1×10 16 , 1×10 17 or higher viral particles, infectious viral units or plaque forming units, and various values ​​and ranges between the above values ​​may also be included therein. Preferably, the oncolytic virus administration dose is 1×10 5 Up to 1×10 10 More preferably, the oncolytic virus can be expressed in an amount equal to or greater than 1×10 5 and less than 1×10 9 pfu doses were administered.

[0167] Oncolytic viruses can be administered parenterally, and such administration can be carried out by any suitable method, such as intratumoral, intraperitoneal, subcutaneous, intradermal, intersegmental or intravenous administration. Among them, intratumoral, intraperitoneal or intravenous administration is preferred.

[0168] Regarding the route of administration, dosage and frequency of administration, the oncolytic virus can be administered to the patient in a variety of ways and quantities, depending on the patient's condition and the presence or absence of side effects. And those skilled in the art can select the best route of administration, dosage and frequency of administration within a suitable range. In addition, the oncolytic virus can be administered in combination with other drugs or physiologically active substances known for the therapeutic effect of the treated disease, or can be administered with other drugs in the form of a combined preparation. Specifically, the oncolytic virus can be provided in the form of an injection.

[0169] In another aspect of the present invention, an antibody or fragment thereof that binds to protein A56 or a fragment thereof is provided. The antibody or fragment thereof is as described in the above-mentioned anticancer agent.

[0170] The antibody can be prepared by a method for preparing an antibody, comprising the steps of: i) administering protein A56 or a fragment thereof to an individual; ii) obtaining B cells from the individual; and iii) culturing the B cells to obtain a recombinant anti-protein A56 antibody.

[0171] In addition, protein A56 or a fragment thereof can be prepared by a method for preparing a protein, comprising the following steps: i) transfecting the vector of the present invention into a host cell; ii) culturing the transfected host cell; and iii) obtaining recombinant protein A56 from the culture.

[0172] The host cell can be a prokaryotic cell or a eukaryotic cell. Specifically, the prokaryotic cell can be an Escherichia coli or a yeast cell. The eukaryotic cell can be an NS / 0 myeloma cell, a 293 cell, a Chinese hamster ovary cell (CHO cell), a HeLa cell, a CapT cell (a cell derived from human amniotic fluid), or a COS cell.

[0173] Transfection can be performed by transient transfection, microinjection, transduction, cell fusion, calcium phosphate precipitation, liposome-mediated transfection, DEAE-dextran-mediated transfection, polybrene-mediated transfection, electroporation, or a gene gun.

[0174] In another aspect of the present invention, a cancer targeting polypeptide or a fragment thereof is provided, the polypeptide comprising an amino acid sequence shown by SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079. The polypeptide or fragment thereof comprising an amino acid sequence shown by SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079 is as described in the above-mentioned anticancer agent.

[0175] In another aspect of the present invention, an isolated nucleic acid encoding a cancer targeting polypeptide or a fragment thereof is provided. The nucleic acid may include a nucleotide sequence shown in SEQ ID NO: 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1072, 1074, 1076 or 1078.

[0176] In yet another aspect of the present invention, a kit for treating cancer is provided, comprising an oncolytic virus and an antibody or a fragment thereof.

[0177] The oncolytic virus and the antibody or fragment thereof are as described above for the anticancer agents.

[0178] Individual refers to an individual suffering from or suffering from a disease that can be alleviated, inhibited or treated by administering an oncolytic virus and an antibody or a fragment thereof. The individual can be a mammal, including a human, and can be a non-human animal. The term "non-human animal" refers to any vertebrate, and can include mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cattle, chickens, amphibians and reptiles. In the case where the oncolytic virus is contained in a first composition for cancer cell killing and targeting and is administered to an individual, the oncolytic virus mainly kills only cancer cells in a specific manner; cancer cells that survive the oncolytic virus express A56 protein on the cell surface, which makes targeted secondary anticancer therapy possible.

[0179] In case the antibody or fragment thereof is contained in a second composition for secondary anti-cancer treatment and administered to an individual, the antibody or fragment thereof can secondarily kill only cancer cells expressing protein A56 on the cell surface.

[0180] The first composition and the second composition can be administered in a pharmaceutically effective amount to treat cancer cells or their metastasis, or to inhibit cancer growth. The pharmaceutically effective amount may vary depending on various factors such as the type of cancer, the age and weight of the patient, the nature and severity of the symptoms, the type of current treatment, the number of treatments, the dosage form and route, and can be easily determined by experts in the relevant field.

[0181] The first composition and the second composition can be administered parenterally, and such administration can be carried out by any suitable method, such as intratumoral, intraperitoneal, subcutaneous, intradermal, intranodal or intravenous administration. Among them, intratumoral, intraperitoneal or intravenous administration is preferred. On the other hand, the dosage of the pharmaceutical composition can be determined according to the dosage regimen, dosage, health status of the patient, etc.

[0182] In another aspect of the present invention, a pharmaceutical composition for targeting cancer is provided, comprising an oncolytic virus. The oncolytic virus is as described in the above-mentioned anticancer agent.

[0183] In another aspect of the present invention, there is provided a use of an oncolytic virus for targeting cancer. In another aspect of the present invention, there is provided a method for targeting cancer, comprising the step of administering an oncolytic virus to an individual. The oncolytic virus is as described in the above-mentioned anticancer agent.

[0184] In another aspect of the present invention, there is provided a use of an oncolytic virus and an antibody or a fragment thereof in treating cancer. The oncolytic virus and the antibody or a fragment thereof are as described in the above-mentioned anticancer agent.

[0185] In another aspect of the present invention, a method for treating cancer is provided, comprising the following steps: i) administering an oncolytic virus to an individual; and ii) administering an antibody or a fragment thereof to an individual. The oncolytic virus and the antibody or a fragment thereof are as described above in the anticancer agent.

[0186] Invention method

[0187] Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are only for illustrative purposes, and the scope of the present invention is not limited thereto.

[0188] I. Preparation of oncolytic viruses containing nucleic acids encoding protein A56 or its fragments and identification of their expression on the surface of tumor cells

[0189] Preparation Example 1.1 Preparation and identification of protein A56 variants

[0190] Wild-type vaccinia virus (New York City Department of Health strain, VR-1536) was purchased from the American Type Culture Collection (ATCC). For recombination, pUC57amp+ (GENEWIZ, USA) carrying the HSV1-TK gene (pSE / L promoter) and the firefly luciferase reporter gene (p7.5 promoter) was used as a shuttle plasmid vector.

[0191] To obtain recombinant viruses, HeLa cells (ATCC) were cultured at 4 × 10 5 Cells were seeded in 6-well plates and then prepared in EMEM medium containing 10% fetal bovine serum. Wild-type vaccinia virus was used to treat at an MOI of 0.05. After 2 hours, the medium was replaced with EMEM medium containing 2% fetal bovine serum and then Xfect was used. TM Polymer (Clonetech 631317, USA) was used to transfer 4 μg of linearized shuttle plasmid vector into cells. Culture was continued for 4 hours. Subsequently, the culture medium was replaced with EMEM medium containing 2% fetal bovine serum and culture was continued for 72 hours. By detecting the luciferase activity in HeLa cells, the recombinant vaccinia virus containing the HSV1-TK gene was obtained.

[0192] Then, mutation of HSV1-TK was induced by 10 consecutive subcultures under the condition of applying a biochemical environment (TK-selective pressure), which selected cells lacking TK function in the presence of BrdU (thymidine analog, 15 μg / ml) in the TK-osteosarcoma (143TK-) cell line. A request for amino acid sequencing of the mutant vaccinia virus was submitted to Macrogen (Seoul, South Korea). As a result, the 46th amino acid at the C-terminus of HSV1-TK of the mutant vaccinia virus was determined, and the codon (caa) encoding glutamine (Gln) was point mutated to a stop codon. In addition, the deletion of the amino acid residue after the 46th position of the C-terminus of HSV1-TK of the mutant vaccinia virus was confirmed. Finally, a mutant vaccinia virus (OTS-412) expressing the HSV1-TK fragment with genetic stability was obtained.

[0193] In addition, the most reported mutations in HSV-TK are frameshift mutations caused by base insertion or deletion, which occur in the nucleotide sequence portions from 430 to 436 (7Gs) and 548 to 583 (6Cs). After wild-type HSV-TK is inserted into vaccinia virus, 98% or more of the mutations occur in these portions. Therefore, in order to produce silent mutations in the nucleic acid sequence portion, the nucleic acid sequence GGGGGGGGG at positions 430 to 436 is changed to GGTGGGTG, and the nucleic acid sequence CCCCCC at positions 548 to 583 is changed to CCCCTC. In addition, the HSV-TK variant gene, in which the alanine at position 167 in the amino acid sequence of HSV-TK is transformed to encode tyrosine, is used as a shuttle vector. Then, the shuttle vector and the Western Reserve strain (WOTS-418) vaccinia virus are used to produce recombinant vaccinia virus in the same manner as OTS-412.

[0194] A request was made to Macrogen for gene sequencing of protein A56 in OTS-412 and WOTS-418. The protein A56 sequences in OTS-412 and WOTS-418 were compared using NCBI Blast and Uniprot. As a result, no 100% identical sequences were found; in particular, a deletion of amino acids at positions 245 to 250 was confirmed. Comparisons with 4 other strains with high sequence homology were performed ( Fig.99 ).

[0195] Preparation Example 1.2. Construction of an oncolytic virus vector containing a nucleic acid encoding protein A56 or a fragment thereof

[0196] Wild-type poxvirus (New York City Department of Health Laboratory) was purchased from the American Type Culture Collection (ATCC; VR-1536). In this virus, the thymidine kinase gene was knocked out by homologous recombination and replaced with a reporter gene and a gene encoding protein A56 or a fragment thereof (SEQ ID NO: 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059 or 1061, 1072, 1074 (OTS-412-A56), 1076, 1078 (WOTS-418-A56).

[0197] Preparation Example 1.3. Preparation of oncolytic viruses containing nucleic acids encoding protein A56 or fragments thereof In order to prepare oncolytic viruses containing protein A56, HeLaS3 (ATCC) cells were cultured at 4×10 5The cells were seeded in a 6-well plate and then prepared in EMEM medium containing 10% fetal bovine serum. Wild-type vaccinia virus was used for treatment at an MOI of 0.05. After 2 hours, EMEM medium containing 2% fetal bovine serum was replaced, and then 4 μg of the oncolytic virus vector constructed in Preparation Example 1.1 was transfected into the cells with Xfect reagent buffer. Culture for 4 hours. Subsequently, the culture medium was replaced with EMEM medium containing 2% fetal bovine serum and cultured for another 72 hours. Finally, the infected cells were collected and frozen and thawed three times. The cells were lysed by ultrasound, and free oncolytic viruses containing nucleic acids encoding protein A56 or fragments thereof were obtained using the sucrose buffer method.

[0198] Experimental Example 1. Expression of protein A56 on the surface of cancer cells

[0199] The oncolytic virus containing the nucleic acid encoding protein A56 or a fragment thereof prepared in Preparation Example 1.3 was used to infect a human lung cancer cell line (A549) or a human colorectal cancer cell line (HCT-116) to determine whether protein A56 is expressed on the surface of cancer cells.

[0200] Specifically, A549 (lung cancer, ATCC, USA) or HCT-116 (colorectal cancer, Korean cell line bank) cell lines were cultured at 3.5 × 10 4 Cells / well were seeded on coverslips in 12-well plates. Subsequently, the cells were infected with oncolytic viruses at an MOI of 0.1 and then incubated for 30 hours at 37 ° C and 5% CO2. Each incubated cell line was harvested. Then, it was treated with 4% (v / v) paraformaldehyde (PFA), 1% (v / v) BSA, an anti-A56 primary antibody diluted at a ratio of 1:500 (Cat. No. ABIN1606294, Antibody Online), and a secondary antibody diluted at a ratio of 1:200 (Alexa 594, Cat. No. A21205, Invitrogen). DAPI staining was performed. Then, samples of each cell line were placed on a slide and observed using a confocal microscope (Olympus, FV1000).

[0201] As a result, it was determined that protein A56 was expressed on the cell surface of A549 and HCT-116 cell lines infected with oncolytic viruses ( Figure 2 and 3 ).

[0202] Experimental Example 2. Identification of the expression of surface protein A56 in mouse tumor tissue (I)

[0203] The oncolytic virus containing the nucleic acid encoding protein A56 or a fragment thereof prepared in Preparation Example 1.3 was intraperitoneally administered to a cancer-induced mouse model to determine whether protein A56 is expressed on the tissue surface.

[0204] Specifically, 6.3×10 6 HT-29 colorectal cancer cell line (Korean Cell Line Bank, KCLB) was subcutaneously transplanted into BALB / c nude mice to induce cancer. When the average tumor volume reached 150mm 3 Up to 200mm 3 When the oncolytic virus prepared in Preparation Example 1.3 was added at 2×10 7 The mice were intraperitoneally administered with a dose of 100 pfu. The mice were then sacrificed on day 4, and tumor tissues and brain, heart, lung, muscle, kidney, liver, and spleen tissues were collected from them.

[0205] Immunofluorescence staining of protein A56 was performed in the same manner as in Experimental Example 1. DAPI staining was performed. Then, a sample of each tissue was placed on a slide and observed using a fluorescence microscope.

[0206] As a result, it was determined that protein A56 was expressed on the surface of tumor tissues of mice treated with oncolytic viruses. On the other hand, it was found that protein A56 was not expressed in brain, heart, lung, muscle, kidney, liver and spleen tissues ( Figure 4 ). From these results, it can be determined that, in the case where an antibody targeting protein A56 expressed on the surface of tumor tissue is produced after oncolytic virus administration, the antibody can be used as an anticancer immunotherapy.

[0207] Experimental Example 3. Identification of the expression of rabbit tissue surface protein A56

[0208] The oncolytic virus containing the nucleic acid encoding protein A56 or a fragment thereof prepared in Preparation Example 1.3 was intravenously administered to normal rabbits to determine whether protein A56 was expressed on the tissue surface.

[0209] Specifically, 1×10 8 pfu or 1×10 9 The oncolytic virus produced in Preparation Example 1.3 was intravenously administered to New Zealand rabbits at a dose of 1.5 pfu. Then, at week 3 or week 8, the rabbits were sacrificed, and brain, heart, lung, muscle, kidney, liver, and spleen tissues were collected therefrom.

[0210] Immunofluorescence staining of protein A56 was performed in the same manner as in Experimental Example 1. DAPI staining was performed. Then, a sample of each tissue was placed on a slide and observed using a fluorescence microscope.

[0211] The results showed that protein A56 was not expressed in the heart, lung, muscle, kidney, liver and spleen tissues of rabbits administered with oncolytic virus.

[0212] On the other hand, a fluorescent reaction was detected in the brain tissue of the rabbits administered with the oncolytic virus. To identify whether the detected fluorescent reaction was a nonspecific reaction of the anti-A56 antibody, the brain tissue of normal rabbits not administered with the oncolytic virus was immunofluorescently stained in the same manner as above, and then the fluorescent reaction therein was examined using a fluorescence microscope.

[0213] As a result, even in the brain tissue of rabbits that were not given oncolytic virus, a fluorescent reaction was detected. From this result, it can be determined that this fluorescent reaction is a nonspecific reaction ( Figure 5 ).

[0214] In addition, for normal human brain tissue (Pusan ​​National University Yangsan Hospital, South Korea) that was not given oncolytic virus, immunofluorescence staining was performed in the same manner as above, and a weak fluorescence reaction was detected using a fluorescence microscope. However, based on the view that antibodies do not cross the blood-brain barrier, it can be determined that anti-A56 antibodies do not cause nonspecific reactions unless the antibodies are directly administered to the ventricles.

[0215] Experimental Example 4. Identification of the expression of surface protein A56 in mouse tumor tissue (II)

[0216] The oncolytic virus containing the nucleic acid encoding protein A56 or a fragment thereof prepared in Preparation Example 1.3 and hydroxyurea were co-administered to a cancer-induced mouse model to determine whether protein A56 was expressed on the surface of its tissues.

[0217] Specifically, 6.3×10 6 Renca cancer cell line (Korean Cell Line Bank) cells were subcutaneously transplanted into BALB / c nude mice to induce cancer. When the average tumor volume reached 150mm 3 ~200mm 3 When the oncolytic virus prepared in Preparation Example 1.3 was added at 2×10 7 The mice were intratumorally administered with a dose of 100 pfu and hydroxyurea at a dose of 30 mg / kg.

[0218] The prepared renal cancer cell transplanted mice were divided into three groups (n=4). The normal saline intratumoral administration group was used as the control group, and the oncolytic virus single administration group (1×10 7 pfu) and oncolytic viruses (1×10 7 The experimental group consisted of a group of 100 mg / kg (pfu) and hydroxyurea (30 mg / kg). The oncolytic virus was administered intratumorally twice on days 0 and 14, and hydroxyurea was administered intraperitoneally 6 times a week from 1 day before oncolytic virus administration to day 21 after oncolytic virus administration, except for the day of oncolytic virus administration.

[0219] Mice were killed on the 7th, 10th and 14th days after the first administration of oncolytic virus, and tumor tissues were collected therefrom. In addition, mice were killed on the 21st, 24th and 28th days after the second administration of oncolytic virus, and tumor tissues were collected therefrom. Immunofluorescence staining of protein A56 was performed in the same manner as in Experimental Example 1. DAPI staining was performed. Then, samples of each tissue were placed on a slide and observed using a fluorescence microscope.

[0220] The results showed that until 7, 10, and 14 days after the first administration of oncolytic virus, the tumor surface of mice in the group receiving oncolytic virus only and the group receiving oncolytic virus and hydroxyurea clearly expressed protein A56 ( Figure 6 and 7 ).

[0221] In addition, it was confirmed that when the second oncolytic virus administration was performed on the 14th day after the first oncolytic virus administration, protein A56 was expressed on the tumor surface until the 7th, 10th and 14th days after the second administration ( Figure 8 and 9 ). In particular, protein A56 was found to be expressed in both dead and living cells on day 24 (D10).

[0222] In addition, it was found that the tumor volume of mice in the group that was given oncolytic virus alone and the group that was given oncolytic virus and hydroxyurea at the same time was significantly reduced compared with the mice in the group that received only saline ( Fig.10 In addition, the body weights of the three groups of mice were measured on days 3, 7, 14, 18, and 21. Results No significant weight loss was observed in any of the groups ( Fig.11 ).

[0223] Experimental Example 5. Expression of protein A56 or its fragments on the cell surface

[0224] The oncolytic virus containing the nucleic acid encoding protein A56 or a fragment thereof prepared in Preparation Example 1.3 was used to treat the A549 cancer cell line to identify whether protein A56 was expressed on the cell surface.

[0225] Specifically, A549 (lung cancer, ATCC, USA) cells were plated at 3.5×10 4Cells were seeded on coverslips in 12-well plates. Subsequently, the cells were infected with an oncolytic virus at an MOI of 0.1 and then incubated for 30 hours at 37 ° C and 5% CO2. Each incubated cell line was harvested. Then, it was treated with 4% (v / v) paraformaldehyde (PFA), 1% (v / v) BSA, anti-A56 antibody diluted at a ratio of 1:500 (Cat. No. ABIN1606294, Antibody Online), and secondary antibody diluted at a ratio of 1:200 (Alexa594, Cat. No. A21205, Invitrogen). The nucleus was stained with DAPI and the Golgi apparatus was stained with fluorescent dyes. Thereafter, the cell line samples were placed on a slide and observed using a confocal microscope (Olympus, FV1000). The results are shown in Fig.12 and 13 and Table 2.

[0226] [Table 2]

[0227]

[0228] Wild-type A56 (A56-G) and the A56 fragment obtained by six regions of partially truncated A56 are allowed to be expressed on the cell surface. The result confirmed that only wild-type A56 (A56-G) and the truncated A56-121 of IgV-like domains reach the cell surface and express thereon. It has been determined that in the A56 variant excluding the IgV-like domain, the variants that the signal peptide, transmembrane domain, stem region and tandem repeat region are truncated individually or in combination are not expressed on the plasma membrane. In addition, it is found that in the case of comprising the IgV-like domain, even the variant that only comprises the transmembrane domain is not expressed on the plasma membrane. That is, it is determined that the variant excluding the IgV-like domain and including five regions is expressed on the plasma membrane.

[0229] II. Generation of Antibodies Binding to Protein A56 or Fragments thereof

[0230] Preparation Example 2. Preparation of protein A56 and its fragments

[0231] Primers were prepared to remove a specific region from the wild-type protein A56, and overlapping PCR was performed by forming a region overlapping with GFP.

[0232] Specifically, the vector DNA (N293F-A56-C-HIS) was amplified and introduced into HEK293F cells for overexpression. Subsequently, primary purification was performed by affinity chromatography (Ni-NTA), and secondary purification was performed by cation exchange chromatography (CEX). In this way, the A56-C-HIS protein ( Fig.97 and 98 ).

[0233] Preparation Example 3. Preparation of Antibodies Binding to Protein A56 or Its Fragments (I)

[0234] Ybiologics has applied for the preparation of anti-A56 antibodies that specifically bind to the A56 protein or its variants or fragments. 61 antibodies were produced using phage library technology and their CDRs were analyzed. The phage library was added to a test tube coated with the A56 antigen, and biopanning was performed to find binding hits. Phages with specific binding were selected by washing 3 times on average. Three screening processes were performed. Then, affinity tests were performed, and a small number of colonies showing high affinity were selected to determine whether the colonies showed affinity with the actual antigen. A collection with a relatively high number of hits was selected, and an automated system was used for selection and hit selection.

[0235] The affinity between each anti-A56 antibody thus produced and protein A56 was measured. The results are shown in Figures 14 to 19 In addition, the yield of each anti-A56 antibody is shown in Figures 20A to 20C. In addition, Figures 21 to 37 The results obtained by identifying each of the produced anti-A56 antibodies by SDS-PAGE are shown.

[0236] Experimental Example 6. Affinity determination of anti-A56 antibody

[0237] Each well of the immunotube was coated with protein A56 and then subjected to a blocking process. After the blocking process, each well was reacted with an antibody serially diluted three times starting from 100 nM and reacted at room temperature for a predetermined time. After that, after washing 3 times with PBS, the secondary antibody was also treated at room temperature for a predetermined time. Subsequently, the affinity between each anti-A56 antibody and protein A56 was measured at each concentration.

[0238] Experimental Example 7. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies (I)

[0239] HeLa, HT-29, A549, MCF-7 and PC-3 cell lines were infected with the oncolytic virus prepared in Preparation Example 1.3 at an MOI of 1. After 24 hours, each cell line infected with the oncolytic virus was treated with 5 anti-A56 antibodies (ab01, ab02, ab03, ab04, ab05) prepared in Preparation Example 3, and then the reaction was allowed to proceed for 1 hour at 4 ° C. Then, it was washed 3 times with PBS and then treated with anti-hIgG antibody (FITC, Abcam). Subsequently, the reaction was allowed to proceed for 1 hour at a temperature of 4 ° C. After 1 hour, the PI staining kit (BD Annexin V kit) was used for staining according to the manufacturer's manual and then analyzed with a flow cytometer (Moflo Astrios EQ, Beckman Coulter).

[0240] like Fig.38 and 39 As shown, the analysis results (blue area) of the cancer cell line (cells only) not treated with oncolytic virus show that the cancer cell line (cells + VV) treated with oncolytic virus binds to the anti-A56 antibody, resulting in a shift to the red area. From these results, it can be determined that the anti-A56 antibody binds to the protein A56 expressed on the surface of tumor cells.

[0241] Preparation Example 4. Preparation of antibodies binding to protein A56 or its fragments (II)

[0242] In addition, an application was submitted to LakePharma for the production of antibodies based on the anti-A56 antibody (Cat. No. ABIN1606294, Antibody Online), and the antibodies were produced by reverse coding engineering. The two antibodies thus produced were named "LakePharma Variant 3" and "LakePharma Variant 4".

[0243] Experimental Example 8. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies (II)

[0244] The HeLa cell line was infected with the oncolytic virus produced in Preparation Example 1.3 at an MOI of 1. After 24 hours, the HeLa cell line infected with the oncolytic virus was treated with anti-A56 antibody (Cat. No. ABIN1606294, Antibody Online) and two anti-A56 antibodies (LakePharma variants 3 and 4) prepared in Preparation Example 4, respectively, and then reacted at 4 ° C for 1 hour. Then, it was washed 3 times with PBS and then treated with anti-hIgG antibody (FITC, Abcam). Subsequently, the reaction was allowed to proceed at a temperature of 4 ° C for 1 hour. After 1 hour, the PI staining kit (BD Annexin V kit) was used for staining according to the manufacturer's manual and then analyzed with a flow cytometer (Moflo Astrios EQ, Beckman Coulter).

[0245] The results showed that, in the case of treatment with anti-A56 antibody (Cat. No. ABIN1606294, Antibody Online) as a positive control (cells + OV), the anti-A56 antibody bound to A56 expressed on the cell surface of the HeLa cell line due to treatment with the oncolytic virus, resulting in a rightward shift ( ) compared to the case of treatment with only the HeLa cell line (cells only). Fig.40 ).

[0246] In addition, in the experimental group (cells + OV), when treated with two anti-A56 antibodies (LakePharma variants 3 and 4), the anti-A56 antibodies bind to A56 expressed on the cell surface of the HeLa cell line due to oncolytic virus treatment, resulting in a rightward shift ( Fig.41 and 42 ). From these results, it can be determined that the two anti-A56 antibodies (LakePharma variants 3 and 4) produced in Preparation Example 4 specifically bind to A56 expressed on the cell surface ( Fig.43 ).

[0247] Experimental Example 9. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: A549 cancer cell line (I)

[0248] The A549 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1.3, so that protein A56 was expressed in the cell line. Then, whether anti-A56 antibodies (ten anti-human A56 antibodies and two anti-mouse A56 antibodies) bind to protein A56 was identified.

[0249] The A549 cancer cell line was treated with various oncolytic viruses with an MOI of 1. After 24 hours, the cells were treated with ten antibodies prepared in Preparation Example 3 and using human scFv as binding domains (top 10 in binding affinity, i.e., Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16) and two antibodies prepared in Preparation Example 4 (LP variant 03 and LP variant 04). After 1 hour, secondary antibodies (goat anti-human IgG FC, goat anti-mouse IgG H&L) were added. After 1 hour, washing was performed, and the A549 cancer cell line was stained with propidium iodide (PI) and analyzed by flow cytometry.

[0250] The results showed that for the A549 cancer cell line treated with oncolytic virus, 10 human antibodies and 2 mouse antibodies showed 90% or higher similar binding signals ( Figures 44 to 55 ). In addition, slight differences were identified among the 10 human antibodies in terms of the peak values ​​observed in the oncolytic virus-treated A549 cancer cell line, and the differences were not significant ( Fig.56 and 57 ).

[0251] Experimental Example 10. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: A549 cancer cell line (II)

[0252] The A549 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1.3, so that protein A56 was expressed in the cell line. Then, it was identified whether the anti-A56 antibodies prepared in Preparation Examples 3 and 4 (Ybiologics: Ab18 and Ab16, LakePharma: LakePharma variants 3, 4) bind to protein A56.

[0253] The A549 cancer cell line was treated with oncolytic virus and anti-A56 antibody in the same manner as in Experimental Example 9. The A549 cancer cell line was stained with propidium iodide (PI) and analyzed using a flow cytometer (Moflo Astrios EQ, Beckman Coulter).

[0254] The results determined that the anti-A56 antibodies produced by Ybiologics and LakePharma respectively showed 93% or higher similar binding signals for the oncolytic virus-treated A549 cancer cell line ( Figures 58 to 61 ).

[0255] Experimental Example 11. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: PC3 cancer cell line (I)

[0256] The PC3 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding the protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1.3, so that the protein A56 was expressed in the cell line. Then, it was identified whether the anti-A56 antibodies (10 human anti-A56 antibodies and 2 mouse anti-A56 antibodies) bind to the A56 protein.

[0257] In the same manner as in Experimental Example 9, PC3 cancer cell lines were treated with various oncolytic viruses, 10 antibodies (top 10 in binding affinity, i.e., Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16) prepared in Preparation Example 3 and using human scFv as binding domains, and two antibodies (LP variant 03 and LP variant 04) prepared in Preparation Example 4. PC3 cancer cell lines were stained with PI and analyzed by flow cytometry (Moflo Astrios EQ, Beckman Coulter).

[0258] The results showed that for PC3 cancer cell lines treated with oncolytic viruses, 10 human antibodies and 2 mouse antibodies showed 90% or higher similar binding signals ( Figures 62 to 73). In addition, there were slight differences among the 10 human antibodies in terms of the peaks observed in the oncolytic virus-treated PC3 cancer cell line, with signals of 60% or higher, among which the anti-A56 antibody Ab51 showed the highest binding signals of 70.71% and 78.61%, respectively ( Fig.74 and 75 ).

[0259] Experimental Example 12. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: PC3 cancer cell line (II)

[0260] The PC3 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1, so that protein A56 was expressed in the cell line. Then, it was identified whether the anti-A56 antibodies prepared in Preparation Examples 3 and 4 (Ybiologics: Ab18 and Ab16, LakePharma: LakePharma variants 3, 4) bind to protein A56.

[0261] The PC3 cancer cell line was treated with oncolytic virus and anti-A56 antibody in the same manner as in Experimental Example 9. The PC3 cancer cell line was stained with PI and analyzed using a flow cytometer (Moflo Astrios EQ, Beckman Coulter).

[0262] The results determined that each anti-A56 antibody (Ab18 and Ab16) produced by Ybiologics showed 60% or more and 70% or more binding signals, respectively, to the oncolytic virus-treated PC3 cancer cell line ( Fig.76 and 77 In addition, it was determined that each anti-A56 antibody produced by LakePharma (LakePharma variants 3, 4) showed a binding signal of 90% or higher ( Fig.78 and 79 ).

[0263] Experimental Example 13. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: MCF7 cancer cell line (I)

[0264] The MCF7 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1, so that protein A56 was expressed in the cell line. Then, it was identified whether anti-A56 antibodies (10 human anti-A56 antibodies and 2 mouse anti-A56 antibodies) bind to the A56 protein.

[0265] In the same manner as in Experimental Example 9, the MCF7 cancer cell line was treated with various oncolytic viruses, ten antibodies (top 10 in binding affinity, i.e., Ab18, Ab19, Ab01, Ab13, Ab14, Ab08, Ab03, Ab51, Ab55, Ab16) prepared in Preparation Example 3 and using human scFv as a binding domain, and two antibodies (LP variant 03 and LP variant 04) prepared in Preparation Example 4. The MCF7 cancer cell line was stained with PI and analyzed by flow cytometry (Moflo Astrios EQ, Beckman Coulter).

[0266] The results showed that for the MCF7 cancer cell line treated with oncolytic virus, 10 human antibodies and 2 mouse antibodies showed 80% or higher similar binding signals ( Figures 80 to 91 ). In addition, it was confirmed that although there were slight differences in the peak values ​​observed in the oncolytic virus-treated MCF7 cancer cell line among the 10 human antibodies, the signals were 80% or higher ( Fig.92 and 93 ).

[0267] Experimental Example 14. Identification of the binding of protein A56 expressed on the surface of tumor cells to anti-A56 antibodies: MCF7 cancer cell line (II)

[0268] The MCF7 cancer cell line was treated with an oncolytic virus (OTS-412 or WOTS-418) encoding protein A56 of SEQ ID NO: 1074 or 1078 and prepared in Preparation Example 1, so that protein A56 was expressed in the cell line. Then, it was identified whether the anti-A56 antibody (LakePharma: LakePharma variants 3, 4) prepared in Preparation Example 4 binds to protein A56.

[0269] The MCF7 cancer cell line was treated with oncolytic virus and anti-A56 antibody in the same manner as in Experimental Example 9. The MCF7 cancer cell line was stained with PI and analyzed using a flow cytometer (Moflo Astrios EQ, Beckman Coulter).

[0270] The results determined that each anti-A56 antibody produced by LakePharma (LakePharma variants 3, 4) showed a binding signal of 97% or higher for the oncolytic virus-treated MCF7 cancer cell line ( Fig.94 and 95 ).

[0271] III. Identification of anticancer effects using oncolytic viruses and antibodies

[0272] Experimental Example 15. Identification of anticancer effects using oncolytic viruses and antibodies

[0273] The oncolytic virus prepared in Preparation Example 1.3 was administered to cancer-induced mice prepared in the same manner as in Experimental Example 2. After a predetermined time, the cancer-induced mice as the control group were treated with saline, and the cancer-induced mice as the experimental group were treated with the antibody produced in Preparation Example 3. It was found that the cancer cells that were not killed by the oncolytic virus remained in the tumor tissue of the control group, while the cancer cells in the tumor tissue of the experimental group were completely killed.

[0274] Experimental Example 16. Identification of anticancer effects using oncolytic viruses and immune cells

[0275] The oncolytic virus prepared in Preparation Example 1.3 was administered to cancer-induced mice prepared in the same manner as in Experimental Example 2. After a predetermined time, the cancer-induced mice as the control group were treated with saline, and the cancer-induced mice as the experimental group were treated with the prepared immune cells. It was found that the cancer cells that were not killed by the oncolytic virus remained in the tumor tissue of the control group, while the cancer cells in the tumor tissue of the experimental group were completely killed.

Claims

1. An anticancer agent targeting cancer cells infected by an oncolytic virus, comprising an active ingredient: An antibody or fragment thereof that specifically binds to protein A56 or a fragment thereof; wherein the oncolytic virus contains a nucleic acid encoding protein A56 or a fragment thereof, and wherein the protein A56 or a fragment thereof comprises a polypeptide having at least 90% sequence homology to the amino acid sequence shown in SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079; The antibody or fragment thereof has: The heavy chain CDR1 of SEQ ID NO: 1; the heavy chain CDR2 of SEQ ID NO: 2; the heavy chain CDR3 of SEQ ID NO: 3; the light chain CDR1 of SEQ ID NO: 4; the light chain CDR2 of SEQ ID NO: 5; and the light chain CDR3 of SEQ ID NO: 6; The heavy chain CDR1 of SEQ ID NO: 18; the heavy chain CDR2 of SEQ ID NO: 19; the heavy chain CDR3 of SEQ ID NO: 20; the light chain CDR1 of SEQ ID NO: 21; the light chain CDR2 of SEQ ID NO: 22; and the light chain CDR3 of SEQ ID NO: 23; The heavy chain CDR1 of SEQ ID NO:35; the heavy chain CDR2 of SEQ ID NO:36; the heavy chain CDR3 of SEQ ID NO:37; the light chain CDR1 of SEQ ID NO:38; the light chain CDR2 of SEQ ID NO:39; and the light chain CDR3 of SEQ ID NO:40; The heavy chain CDR1 of SEQ ID NO:52; the heavy chain CDR2 of SEQ ID NO:53; the heavy chain CDR3 of SEQ ID NO:54; the light chain CDR1 of SEQ ID NO:55; the light chain CDR2 of SEQ ID NO:56; and the light chain CDR3 of SEQ ID NO:57; The heavy chain CDR1 of SEQ ID NO:69; the heavy chain CDR2 of SEQ ID NO:70; the heavy chain CDR3 of SEQ ID NO:71; the light chain CDR1 of SEQ ID NO:72; the light chain CDR2 of SEQ ID NO:73; and the light chain CDR3 of SEQ ID NO:74; The heavy chain CDR1 of SEQ ID NO:86; the heavy chain CDR2 of SEQ ID NO:87; the heavy chain CDR3 of SEQ ID NO:88; the light chain CDR1 of SEQ ID NO:89; the light chain CDR2 of SEQ ID NO:90; and the light chain CDR3 of SEQ ID NO:91; The heavy chain CDR1 of SEQ ID NO: 103; the heavy chain CDR2 of SEQ ID NO: 104; the heavy chain CDR3 of SEQ ID NO: 105; the light chain CDR1 of SEQ ID NO: 106; the light chain CDR2 of SEQ ID NO: 107; and the light chain CDR3 of SEQ ID NO: 108; The heavy chain CDR1 of SEQ ID NO: 120; the heavy chain CDR2 of SEQ ID NO: 121; the heavy chain CDR3 of SEQ ID NO: 122; the light chain CDR1 of SEQ ID NO: 123; the light chain CDR2 of SEQ ID NO: 124; and the light chain CDR3 of SEQ ID NO: 125; The heavy chain CDR1 of SEQ ID NO: 137, the heavy chain CDR2 of SEQ ID NO: 138, the heavy chain CDR3 of SEQ ID NO: 139, the light chain CDR1 of SEQ ID NO: 140, the light chain CDR2 of SEQ ID NO: 141, and the light chain CDR3 of SEQ ID NO: 142; The heavy chain CDR1 of SEQ ID NO: 154, the heavy chain CDR2 of SEQ ID NO: 155, the heavy chain CDR3 of SEQ ID NO: 156, the light chain CDR1 of SEQ ID NO: 157, the light chain CDR2 of SEQ ID NO: 158, and the light chain CDR3 of SEQ ID NO: 159; The heavy chain CDR1 of SEQ ID NO: 171, the heavy chain CDR2 of SEQ ID NO: 172, the heavy chain CDR3 of SEQ ID NO: 173, the light chain CDR1 of SEQ ID NO: 174, the light chain CDR2 of SEQ ID NO: 175, and the light chain CDR3 of SEQ ID NO: 176; The heavy chain CDR1 of SEQ ID NO: 188, the heavy chain CDR2 of SEQ ID NO: 189, the heavy chain CDR3 of SEQ ID NO: 190, the light chain CDR1 of SEQ ID NO: 191, the light chain CDR2 of SEQ ID NO: 192, and the light chain CDR3 of SEQ ID NO: 193; The heavy chain CDR1 of SEQ ID NO:205, the heavy chain CDR2 of SEQ ID NO:206, the heavy chain CDR3 of SEQ ID NO:207, the light chain CDR1 of SEQ ID NO:208, the light chain CDR2 of SEQ ID NO:209, and the light chain CDR3 of SEQ ID NO:210; The heavy chain CDR1 of SEQ ID NO:222; the heavy chain CDR2 of SEQ ID NO:223; the heavy chain CDR3 of SEQ ID NO:224; the light chain CDR1 of SEQ ID NO:225; the light chain CDR2 of SEQ ID NO:226; and the light chain CDR3 of SEQ ID NO:227; The heavy chain CDR1 of SEQ ID NO:239; the heavy chain CDR2 of SEQ ID NO:240; the heavy chain CDR3 of SEQ ID NO:241; the light chain CDR1 of SEQ ID NO:242; the light chain CDR2 of SEQ ID NO:243; and the light chain CDR3 of SEQ ID NO:244; The heavy chain CDR1 of SEQ ID NO:256, the heavy chain CDR2 of SEQ ID NO:257, the heavy chain CDR3 of SEQ ID NO:258, the light chain CDR1 of SEQ ID NO:259, the light chain CDR2 of SEQ ID NO:260, and the light chain CDR3 of SEQ ID NO:261; The heavy chain CDR1 of SEQ ID NO:273, the heavy chain CDR2 of SEQ ID NO:274, the heavy chain CDR3 of SEQ ID NO:275, the light chain CDR1 of SEQ ID NO:276, the light chain CDR2 of SEQ ID NO:277, and the light chain CDR3 of SEQ ID NO:278; The heavy chain CDR1 of SEQ ID NO:290; the heavy chain CDR2 of SEQ ID NO:291; the heavy chain CDR3 of SEQ ID NO:292; the light chain CDR1 of SEQ ID NO:293; the light chain CDR2 of SEQ ID NO:294; and the light chain CDR3 of SEQ ID NO:295; The heavy chain CDR1 of SEQ ID NO:307, the heavy chain CDR2 of SEQ ID NO:308, the heavy chain CDR3 of SEQ ID NO:309, the light chain CDR1 of SEQ ID NO:310, the light chain CDR2 of SEQ ID NO:311, and the light chain CDR3 of SEQ ID NO:312; The heavy chain CDR1 of SEQ ID NO:324; the heavy chain CDR2 of SEQ ID NO:325; the heavy chain CDR3 of SEQ ID NO:326; the light chain CDR1 of SEQ ID NO:327; the light chain CDR2 of SEQ ID NO:328; and the light chain CDR3 of SEQ ID NO:329; The heavy chain CDR1 of SEQ ID NO:341; the heavy chain CDR2 of SEQ ID NO:342; the heavy chain CDR3 of SEQ ID NO:343; the light chain CDR1 of SEQ ID NO:344; the light chain CDR2 of SEQ ID NO:345; and the light chain CDR3 of SEQ ID NO:346; The heavy chain CDR1 of SEQ ID NO:358, the heavy chain CDR2 of SEQ ID NO:359, the heavy chain CDR3 of SEQ ID NO:360, the light chain CDR1 of SEQ ID NO:361, the light chain CDR2 of SEQ ID NO:362, and the light chain CDR3 of SEQ ID NO:363; The heavy chain CDR1 of SEQ ID NO:375, the heavy chain CDR2 of SEQ ID NO:376, the heavy chain CDR3 of SEQ ID NO:377, the light chain CDR1 of SEQ ID NO:378, the light chain CDR2 of SEQ ID NO:379, and the light chain CDR3 of SEQ ID NO:380; The heavy chain CDR1 of SEQ ID NO:392, the heavy chain CDR2 of SEQ ID NO:393, the heavy chain CDR3 of SEQ ID NO:394, the light chain CDR1 of SEQ ID NO:395, the light chain CDR2 of SEQ ID NO:396, and the light chain CDR3 of SEQ ID NO:397; The heavy chain CDR1 of SEQ ID NO:409, the heavy chain CDR2 of SEQ ID NO:410, the heavy chain CDR3 of SEQ ID NO:411, the light chain CDR1 of SEQ ID NO:412, the light chain CDR2 of SEQ ID NO:413, and the light chain CDR3 of SEQ ID NO:414; The heavy chain CDR1 of SEQ ID NO:426, the heavy chain CDR2 of SEQ ID NO:427, the heavy chain CDR3 of SEQ ID NO:428, the light chain CDR1 of SEQ ID NO:429, the light chain CDR2 of SEQ ID NO:430, and the light chain CDR3 of SEQ ID NO:431; The heavy chain CDR1 of SEQ ID NO:443, the heavy chain CDR2 of SEQ ID NO:444, the heavy chain CDR3 of SEQ ID NO:445, the light chain CDR1 of SEQ ID NO:446, the light chain CDR2 of SEQ ID NO:447, and the light chain CDR3 of SEQ ID NO:448; The heavy chain CDR1 of SEQ ID NO:460, the heavy chain CDR2 of SEQ ID NO:461, the heavy chain CDR3 of SEQ ID NO:462, the light chain CDR1 of SEQ ID NO:463, the light chain CDR2 of SEQ ID NO:464, and the light chain CDR3 of SEQ ID NO:465; The heavy chain CDR1 of SEQ ID NO:477, the heavy chain CDR2 of SEQ ID NO:478, the heavy chain CDR3 of SEQ ID NO:479, the light chain CDR1 of SEQ ID NO:480, the light chain CDR2 of SEQ ID NO:481, and the light chain CDR3 of SEQ ID NO:482; The heavy chain CDR1 of SEQ ID NO:494, the heavy chain CDR2 of SEQ ID NO:495, the heavy chain CDR3 of SEQ ID NO:496, the light chain CDR1 of SEQ ID NO:497, the light chain CDR2 of SEQ ID NO:498, and the light chain CDR3 of SEQ ID NO:499; The heavy chain CDR1 of SEQ ID NO:511, the heavy chain CDR2 of SEQ ID NO:512, the heavy chain CDR3 of SEQ ID NO:513, the light chain CDR1 of SEQ ID NO:514, the light chain CDR2 of SEQ ID NO:515, and the light chain CDR3 of SEQ ID NO:516; The heavy chain CDR1 of SEQ ID NO:528, the heavy chain CDR2 of SEQ ID NO:529, the heavy chain CDR3 of SEQ ID NO:530, the light chain CDR1 of SEQ ID NO:531, the light chain CDR2 of SEQ ID NO:532, and the light chain CDR3 of SEQ ID NO:533; The heavy chain CDR1 of SEQ ID NO:545, the heavy chain CDR2 of SEQ ID NO:546, the heavy chain CDR3 of SEQ ID NO:547, the light chain CDR1 of SEQ ID NO:548, the light chain CDR2 of SEQ ID NO:549, and the light chain CDR3 of SEQ ID NO:550; The heavy chain CDR1 of SEQ ID NO:562, the heavy chain CDR2 of SEQ ID NO:563, the heavy chain CDR3 of SEQ ID NO:564, the light chain CDR1 of SEQ ID NO:565, the light chain CDR2 of SEQ ID NO:566, and the light chain CDR3 of SEQ ID NO:567; The heavy chain CDR1 of SEQ ID NO:579, the heavy chain CDR2 of SEQ ID NO:580, the heavy chain CDR3 of SEQ ID NO:581, the light chain CDR1 of SEQ ID NO:582, the light chain CDR2 of SEQ ID NO:583, and the light chain CDR3 of SEQ ID NO:584; The heavy chain CDR1 of SEQ ID NO:596, the heavy chain CDR2 of SEQ ID NO:597, the heavy chain CDR3 of SEQ ID NO:598, the light chain CDR1 of SEQ ID NO:599, the light chain CDR2 of SEQ ID NO:600, and the light chain CDR3 of SEQ ID NO:601; The heavy chain CDR1 of SEQ ID NO:613, the heavy chain CDR2 of SEQ ID NO:614, the heavy chain CDR3 of SEQ ID NO:615, the light chain CDR1 of SEQ ID NO:616, the light chain CDR2 of SEQ ID NO:617, and the light chain CDR3 of SEQ ID NO:618; The heavy chain CDR1 of SEQ ID NO:630, the heavy chain CDR2 of SEQ ID NO:631, the heavy chain CDR3 of SEQ ID NO:632, the light chain CDR1 of SEQ ID NO:633, the light chain CDR2 of SEQ ID NO:634, and the light chain CDR3 of SEQ ID NO:635; The heavy chain CDR1 of SEQ ID NO:647, the heavy chain CDR2 of SEQ ID NO:648, the heavy chain CDR3 of SEQ ID NO:649, the light chain CDR1 of SEQ ID NO:650, the light chain CDR2 of SEQ ID NO:651, and the light chain CDR3 of SEQ ID NO:652; The heavy chain CDR1 of SEQ ID NO:664, the heavy chain CDR2 of SEQ ID NO:665, the heavy chain CDR3 of SEQ ID NO:666, the light chain CDR1 of SEQ ID NO:667, the light chain CDR2 of SEQ ID NO:668, and the light chain CDR3 of SEQ ID NO:669; The heavy chain CDR1 of SEQ ID NO:681, the heavy chain CDR2 of SEQ ID NO:682, the heavy chain CDR3 of SEQ ID NO:683, the light chain CDR1 of SEQ ID NO:684, the light chain CDR2 of SEQ ID NO:685, and the light chain CDR3 of SEQ ID NO:686; The heavy chain CDR1 of SEQ ID NO:698, the heavy chain CDR2 of SEQ ID NO:699, the heavy chain CDR3 of SEQ ID NO:700, the light chain CDR1 of SEQ ID NO:701, the light chain CDR2 of SEQ ID NO:702, and the light chain CDR3 of SEQ ID NO:703; The heavy chain CDR1 of SEQ ID NO:715, the heavy chain CDR2 of SEQ ID NO:716, the heavy chain CDR3 of SEQ ID NO:717, the light chain CDR1 of SEQ ID NO:718, the light chain CDR2 of SEQ ID NO:719, and the light chain CDR3 of SEQ ID NO:720; The heavy chain CDR1 of SEQ ID NO:732, the heavy chain CDR2 of SEQ ID NO:733, the heavy chain CDR3 of SEQ ID NO:734, the light chain CDR1 of SEQ ID NO:735, the light chain CDR2 of SEQ ID NO:736, and the light chain CDR3 of SEQ ID NO:737; The heavy chain CDR1 of SEQ ID NO:749, the heavy chain CDR2 of SEQ ID NO:750, the heavy chain CDR3 of SEQ ID NO:751, the light chain CDR1 of SEQ ID NO:752, the light chain CDR2 of SEQ ID NO:753, and the light chain CDR3 of SEQ ID NO:754; The heavy chain CDR1 of SEQ ID NO:766, the heavy chain CDR2 of SEQ ID NO:767, the heavy chain CDR3 of SEQ ID NO:768, the light chain CDR1 of SEQ ID NO:769, the light chain CDR2 of SEQ ID NO:770, and the light chain CDR3 of SEQ ID NO:771; The heavy chain CDR1 of SEQ ID NO:783, the heavy chain CDR2 of SEQ ID NO:784, the heavy chain CDR3 of SEQ ID NO:785, the light chain CDR1 of SEQ ID NO:786, the light chain CDR2 of SEQ ID NO:787, and the light chain CDR3 of SEQ ID NO:788; The heavy chain CDR1 of SEQ ID NO:800, the heavy chain CDR2 of SEQ ID NO:801, the heavy chain CDR3 of SEQ ID NO:802, the light chain CDR1 of SEQ ID NO:803, the light chain CDR2 of SEQ ID NO:804, and the light chain CDR3 of SEQ ID NO:805; The heavy chain CDR1 of SEQ ID NO:817, the heavy chain CDR2 of SEQ ID NO:818, the heavy chain CDR3 of SEQ ID NO:819, the light chain CDR1 of SEQ ID NO:820, the light chain CDR2 of SEQ ID NO:821, and the light chain CDR3 of SEQ ID NO:822; The heavy chain CDR1 of SEQ ID NO:834, the heavy chain CDR2 of SEQ ID NO:835, the heavy chain CDR3 of SEQ ID NO:836, the light chain CDR1 of SEQ ID NO:837, the light chain CDR2 of SEQ ID NO:838, and the light chain CDR3 of SEQ ID NO:839; The heavy chain CDR1 of SEQ ID NO:851; the heavy chain CDR2 of SEQ ID NO:852; the heavy chain CDR3 of SEQ ID NO:853; the light chain CDR1 of SEQ ID NO:854; the light chain CDR2 of SEQ ID NO:855; and the light chain CDR3 of SEQ ID NO:856; The heavy chain CDR1 of SEQ ID NO:868, the heavy chain CDR2 of SEQ ID NO:869, the heavy chain CDR3 of SEQ ID NO:870, the light chain CDR1 of SEQ ID NO:871, the light chain CDR2 of SEQ ID NO:872, and the light chain CDR3 of SEQ ID NO:873; The heavy chain CDR1 of SEQ ID NO:885, the heavy chain CDR2 of SEQ ID NO:886, the heavy chain CDR3 of SEQ ID NO:887, the light chain CDR1 of SEQ ID NO:888, the light chain CDR2 of SEQ ID NO:889, and the light chain CDR3 of SEQ ID NO:890; The heavy chain CDR1 of SEQ ID NO:902, the heavy chain CDR2 of SEQ ID NO:903, the heavy chain CDR3 of SEQ ID NO:904, the light chain CDR1 of SEQ ID NO:905, the light chain CDR2 of SEQ ID NO:906, and the light chain CDR3 of SEQ ID NO:907; The heavy chain CDR1 of SEQ ID NO:919; the heavy chain CDR2 of SEQ ID NO:920; the heavy chain CDR3 of SEQ ID NO:921; the light chain CDR1 of SEQ ID NO:922; the light chain CDR2 of SEQ ID NO:923; and the light chain CDR3 of SEQ ID NO:924; The heavy chain CDR1 of SEQ ID NO:936, the heavy chain CDR2 of SEQ ID NO:937, the heavy chain CDR3 of SEQ ID NO:938, the light chain CDR1 of SEQ ID NO:939, the light chain CDR2 of SEQ ID NO:940, and the light chain CDR3 of SEQ ID NO:941; The heavy chain CDR1 of SEQ ID NO:953, the heavy chain CDR2 of SEQ ID NO:954, the heavy chain CDR3 of SEQ ID NO:955, the light chain CDR1 of SEQ ID NO:956, the light chain CDR2 of SEQ ID NO:957, and the light chain CDR3 of SEQ ID NO:958; The heavy chain CDR1 of SEQ ID NO:970, the heavy chain CDR2 of SEQ ID NO:971, the heavy chain CDR3 of SEQ ID NO:972, the light chain CDR1 of SEQ ID NO:973, the light chain CDR2 of SEQ ID NO:974, and the light chain CDR3 of SEQ ID NO:975; The heavy chain CDR1 of SEQ ID NO:987, the heavy chain CDR2 of SEQ ID NO:988, the heavy chain CDR3 of SEQ ID NO:989, the light chain CDR1 of SEQ ID NO:990, the light chain CDR2 of SEQ ID NO:991, and the light chain CDR3 of SEQ ID NO:992; The heavy chain CDR1 of SEQ ID NO: 1004, the heavy chain CDR2 of SEQ ID NO: 1005, the heavy chain CDR3 of SEQ ID NO: 1006, the light chain CDR1 of SEQ ID NO: 1007, the light chain CDR2 of SEQ ID NO: 1008, and the light chain CDR3 of SEQ ID NO: 1009; The heavy chain CDR1 of SEQ ID NO: 1021, the heavy chain CDR2 of SEQ ID NO: 1022, the heavy chain CDR3 of SEQ ID NO: 1023, the light chain CDR1 of SEQ ID NO: 1024, the light chain CDR2 of SEQ ID NO: 1025, and the light chain CDR3 of SEQ ID NO: 1026; The heavy chain CDR1 of SEQ ID NO: 1062; the heavy chain CDR2 of SEQ ID NO: 1064; the heavy chain CDR3 of SEQ ID NO: 1065; the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068; or The heavy chain CDR1 of SEQ ID NO:1063; the heavy chain CDR2 of SEQ ID NO:1064; the heavy chain CDR3 of SEQ ID NO:1065; the light chain CDR1 of SEQ ID NO:1066; the light chain CDR2 of SEQ ID NO:1067; and the light chain CDR3 of SEQ ID NO:1068.

2. The anticancer agent according to claim 1, wherein the antibody has: The heavy chain CDR1 of SEQ ID NO: 1; the heavy chain CDR2 of SEQ ID NO: 2; and the heavy chain CDR3 of SEQ ID NO: 3, the light chain CDR1 of SEQ ID NO: 4; the light chain CDR2 of SEQ ID NO: 5; and the light chain CDR3 of SEQ ID NO: 6; The heavy chain CDR1 of SEQ ID NO:35; the heavy chain CDR2 of SEQ ID NO:36; and the heavy chain CDR3 of SEQ ID NO:37, the light chain CDR1 of SEQ ID NO:38; the light chain CDR2 of SEQ ID NO:39; and the light chain CDR3 of SEQ ID NO:40; The heavy chain CDR1 of SEQ ID NO: 120; the heavy chain CDR2 of SEQ ID NO: 121; and the heavy chain CDR3 of SEQ ID NO: 122, the light chain CDR1 of SEQ ID NO: 123; the light chain CDR2 of SEQ ID NO: 124; and the light chain CDR3 of SEQ ID NO: 125; The heavy chain CDR1 of SEQ ID NO:222; the heavy chain CDR2 of SEQ ID NO:223; and the heavy chain CDR3 of SEQ ID NO:224, the light chain CDR1 of SEQ ID NO:225; the light chain CDR2 of SEQ ID NO:226; and the light chain CDR3 of SEQ ID NO:227; The heavy chain CDR1 of SEQ ID NO:239; the heavy chain CDR2 of SEQ ID NO:240; and the heavy chain CDR3 of SEQ ID NO:241, the light chain CDR1 of SEQ ID NO:242; the light chain CDR2 of SEQ ID NO:243; and the light chain CDR3 of SEQ ID NO:244; The heavy chain CDR1 of SEQ ID NO:290; the heavy chain CDR2 of SEQ ID NO:291; and the heavy chain CDR3 of SEQ ID NO:292, the light chain CDR1 of SEQ ID NO:293; the light chain CDR2 of SEQ ID NO:294; and the light chain CDR3 of SEQ ID NO:295; The heavy chain CDR1 of SEQ ID NO:324; the heavy chain CDR2 of SEQ ID NO:325; and the heavy chain CDR3 of SEQ ID NO:326, the light chain CDR1 of SEQ ID NO:327; the light chain CDR2 of SEQ ID NO:328; and the light chain CDR3 of SEQ ID NO:329; The heavy chain CDR1 of SEQ ID NO:341; the heavy chain CDR2 of SEQ ID NO:342; and the heavy chain CDR3 of SEQ ID NO:343, the light chain CDR1 of SEQ ID NO:344; the light chain CDR2 of SEQ ID NO:345; and the light chain CDR3 of SEQ ID NO:346; The heavy chain CDR1 of SEQ ID NO:851; the heavy chain CDR2 of SEQ ID NO:852; and the heavy chain CDR3 of SEQ ID NO:853, the light chain CDR1 of SEQ ID NO:854; the light chain CDR2 of SEQ ID NO:855; and the light chain CDR3 of SEQ ID NO:856; The heavy chain CDR1 of SEQ ID NO:919; the heavy chain CDR2 of SEQ ID NO:920; and the heavy chain CDR3 of SEQ ID NO:921, the light chain CDR1 of SEQ ID NO:922; the light chain CDR2 of SEQ ID NO:923; and the light chain CDR3 of SEQ ID NO:924; The heavy chain CDR1 of SEQ ID NO: 1062; the heavy chain CDR2 of SEQ ID NO: 1064; and the heavy chain CDR3 of SEQ ID NO: 1065, the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068; or The heavy chain CDR1 of SEQ ID NO: 1063; the heavy chain CDR2 of SEQ ID NO: 1064; and the heavy chain CDR3 of SEQ ID NO: 1065, the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068.

3. The anticancer agent according to claim 1, wherein the oncolytic virus is vaccinia virus. The anticancer agent according to claim 1 , wherein the protein A56 is a wild-type protein A56 or a variant of protein A56.

5. The anticancer agent of claim 1, wherein the protein A56 or a fragment thereof comprises the amino acid sequence shown in SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079.

6. The anticancer agent of claim 1, wherein the nucleic acid encoding protein A56 or a fragment thereof comprises the nucleotide sequence shown in SEQ ID NO: 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1072, 1074, 1076, or 1078.

7. The anticancer agent of claim 1, wherein the anticancer agent is used to prevent or treat a cancer selected from the group consisting of lung cancer, colorectal cancer, prostate cancer, breast cancer, uterine cancer, or a combination thereof.

8. An antibody or fragment thereof that binds to protein A56 or a fragment thereof, wherein the protein A56 or a fragment thereof is contained in an oncolytic virus and the protein A56 or a fragment thereof is expressed on the surface of a cancer cell infected with the oncolytic virus, and wherein the protein A56 or a fragment thereof comprises a polypeptide having at least 90% sequence homology to the amino acid sequence shown in SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079, wherein the antibody has: The heavy chain CDR1 of SEQ ID NO: 1; the heavy chain CDR2 of SEQ ID NO: 2; the heavy chain CDR3 of SEQ ID NO: 3; the light chain CDR1 of SEQ ID NO: 4; the light chain CDR2 of SEQ ID NO: 5; and the light chain CDR3 of SEQ ID NO: 6; The heavy chain CDR1 of SEQ ID NO: 18; the heavy chain CDR2 of SEQ ID NO: 19; the heavy chain CDR3 of SEQ ID NO: 20; the light chain CDR1 of SEQ ID NO: 21; the light chain CDR2 of SEQ ID NO: 22; and the light chain CDR3 of SEQ ID NO: 23; The heavy chain CDR1 of SEQ ID NO:35; the heavy chain CDR2 of SEQ ID NO:36; the heavy chain CDR3 of SEQ ID NO:37; the light chain CDR1 of SEQ ID NO:38; the light chain CDR2 of SEQ ID NO:39; and the light chain CDR3 of SEQ ID NO:40; The heavy chain CDR1 of SEQ ID NO:52; the heavy chain CDR2 of SEQ ID NO:53; the heavy chain CDR3 of SEQ ID NO:54; the light chain CDR1 of SEQ ID NO:55; the light chain CDR2 of SEQ ID NO:56; and the light chain CDR3 of SEQ ID NO:57; The heavy chain CDR1 of SEQ ID NO:69; the heavy chain CDR2 of SEQ ID NO:70; the heavy chain CDR3 of SEQ ID NO:71; the light chain CDR1 of SEQ ID NO:72; the light chain CDR2 of SEQ ID NO:73; and the light chain CDR3 of SEQ ID NO:74; The heavy chain CDR1 of SEQ ID NO:86; the heavy chain CDR2 of SEQ ID NO:87; the heavy chain CDR3 of SEQ ID NO:88; the light chain CDR1 of SEQ ID NO:89; the light chain CDR2 of SEQ ID NO:90; and the light chain CDR3 of SEQ ID NO:91; The heavy chain CDR1 of SEQ ID NO: 103; the heavy chain CDR2 of SEQ ID NO: 104; the heavy chain CDR3 of SEQ ID NO: 105; the light chain CDR1 of SEQ ID NO: 106; the light chain CDR2 of SEQ ID NO: 107; and the light chain CDR3 of SEQ ID NO: 108; The heavy chain CDR1 of SEQ ID NO: 120; the heavy chain CDR2 of SEQ ID NO: 121; the heavy chain CDR3 of SEQ ID NO: 122; the light chain CDR1 of SEQ ID NO: 123; the light chain CDR2 of SEQ ID NO: 124; and the light chain CDR3 of SEQ ID NO: 125; The heavy chain CDR1 of SEQ ID NO: 137, the heavy chain CDR2 of SEQ ID NO: 138, the heavy chain CDR3 of SEQ ID NO: 139, the light chain CDR1 of SEQ ID NO: 140, the light chain CDR2 of SEQ ID NO: 141, and the light chain CDR3 of SEQ ID NO: 142; The heavy chain CDR1 of SEQ ID NO: 154, the heavy chain CDR2 of SEQ ID NO: 155, the heavy chain CDR3 of SEQ ID NO: 156, the light chain CDR1 of SEQ ID NO: 157, the light chain CDR2 of SEQ ID NO: 158, and the light chain CDR3 of SEQ ID NO: 159; The heavy chain CDR1 of SEQ ID NO: 171, the heavy chain CDR2 of SEQ ID NO: 172, the heavy chain CDR3 of SEQ ID NO: 173, the light chain CDR1 of SEQ ID NO: 174, the light chain CDR2 of SEQ ID NO: 175, and the light chain CDR3 of SEQ ID NO: 176; The heavy chain CDR1 of SEQ ID NO: 188, the heavy chain CDR2 of SEQ ID NO: 189, the heavy chain CDR3 of SEQ ID NO: 190, the light chain CDR1 of SEQ ID NO: 191, the light chain CDR2 of SEQ ID NO: 192, and the light chain CDR3 of SEQ ID NO: 193; The heavy chain CDR1 of SEQ ID NO:205, the heavy chain CDR2 of SEQ ID NO:206, the heavy chain CDR3 of SEQ ID NO:207, the light chain CDR1 of SEQ ID NO:208, the light chain CDR2 of SEQ ID NO:209, and the light chain CDR3 of SEQ ID NO:210; The heavy chain CDR1 of SEQ ID NO:222; the heavy chain CDR2 of SEQ ID NO:223; the heavy chain CDR3 of SEQ ID NO:224; the light chain CDR1 of SEQ ID NO:225; the light chain CDR2 of SEQ ID NO:226; and the light chain CDR3 of SEQ ID NO:227; The heavy chain CDR1 of SEQ ID NO:239; the heavy chain CDR2 of SEQ ID NO:240; the heavy chain CDR3 of SEQ ID NO:241; the light chain CDR1 of SEQ ID NO:242; the light chain CDR2 of SEQ ID NO:243; and the light chain CDR3 of SEQ ID NO:244; The heavy chain CDR1 of SEQ ID NO:256, the heavy chain CDR2 of SEQ ID NO:257, the heavy chain CDR3 of SEQ ID NO:258, the light chain CDR1 of SEQ ID NO:259, the light chain CDR2 of SEQ ID NO:260, and the light chain CDR3 of SEQ ID NO:261; The heavy chain CDR1 of SEQ ID NO:273, the heavy chain CDR2 of SEQ ID NO:274, the heavy chain CDR3 of SEQ ID NO:275, the light chain CDR1 of SEQ ID NO:276, the light chain CDR2 of SEQ ID NO:277, and the light chain CDR3 of SEQ ID NO:278; The heavy chain CDR1 of SEQ ID NO:290; the heavy chain CDR2 of SEQ ID NO:291; the heavy chain CDR3 of SEQ ID NO:292; the light chain CDR1 of SEQ ID NO:293; the light chain CDR2 of SEQ ID NO:294; and the light chain CDR3 of SEQ ID NO:295; The heavy chain CDR1 of SEQ ID NO:307, the heavy chain CDR2 of SEQ ID NO:308, the heavy chain CDR3 of SEQ ID NO:309, the light chain CDR1 of SEQ ID NO:310, the light chain CDR2 of SEQ ID NO:311, and the light chain CDR3 of SEQ ID NO:312; The heavy chain CDR1 of SEQ ID NO:324; the heavy chain CDR2 of SEQ ID NO:325; the heavy chain CDR3 of SEQ ID NO:326; the light chain CDR1 of SEQ ID NO:327; the light chain CDR2 of SEQ ID NO:328; and the light chain CDR3 of SEQ ID NO:329; The heavy chain CDR1 of SEQ ID NO:341; the heavy chain CDR2 of SEQ ID NO:342; the heavy chain CDR3 of SEQ ID NO:343; the light chain CDR1 of SEQ ID NO:344; the light chain CDR2 of SEQ ID NO:345; and the light chain CDR3 of SEQ ID NO:346; The heavy chain CDR1 of SEQ ID NO:358, the heavy chain CDR2 of SEQ ID NO:359, the heavy chain CDR3 of SEQ ID NO:360, the light chain CDR1 of SEQ ID NO:361, the light chain CDR2 of SEQ ID NO:362, and the light chain CDR3 of SEQ ID NO:363; The heavy chain CDR1 of SEQ ID NO:375, the heavy chain CDR2 of SEQ ID NO:376, the heavy chain CDR3 of SEQ ID NO:377, the light chain CDR1 of SEQ ID NO:378, the light chain CDR2 of SEQ ID NO:379, and the light chain CDR3 of SEQ ID NO:380; The heavy chain CDR1 of SEQ ID NO:392, the heavy chain CDR2 of SEQ ID NO:393, the heavy chain CDR3 of SEQ ID NO:394, the light chain CDR1 of SEQ ID NO:395, the light chain CDR2 of SEQ ID NO:396, and the light chain CDR3 of SEQ ID NO:397; The heavy chain CDR1 of SEQ ID NO:409, the heavy chain CDR2 of SEQ ID NO:410, the heavy chain CDR3 of SEQ ID NO:411, the light chain CDR1 of SEQ ID NO:412, the light chain CDR2 of SEQ ID NO:413, and the light chain CDR3 of SEQ ID NO:414; The heavy chain CDR1 of SEQ ID NO:426, the heavy chain CDR2 of SEQ ID NO:427, the heavy chain CDR3 of SEQ ID NO:428, the light chain CDR1 of SEQ ID NO:429, the light chain CDR2 of SEQ ID NO:430, and the light chain CDR3 of SEQ ID NO:431; The heavy chain CDR1 of SEQ ID NO:443, the heavy chain CDR2 of SEQ ID NO:444, the heavy chain CDR3 of SEQ ID NO:445, the light chain CDR1 of SEQ ID NO:446, the light chain CDR2 of SEQ ID NO:447, and the light chain CDR3 of SEQ ID NO:448; The heavy chain CDR1 of SEQ ID NO:460, the heavy chain CDR2 of SEQ ID NO:461, the heavy chain CDR3 of SEQ ID NO:462, the light chain CDR1 of SEQ ID NO:463, the light chain CDR2 of SEQ ID NO:464, and the light chain CDR3 of SEQ ID NO:465; The heavy chain CDR1 of SEQ ID NO:477, the heavy chain CDR2 of SEQ ID NO:478, the heavy chain CDR3 of SEQ ID NO:479, the light chain CDR1 of SEQ ID NO:480, the light chain CDR2 of SEQ ID NO:481, and the light chain CDR3 of SEQ ID NO:482; The heavy chain CDR1 of SEQ ID NO:494, the heavy chain CDR2 of SEQ ID NO:495, the heavy chain CDR3 of SEQ ID NO:496, the light chain CDR1 of SEQ ID NO:497, the light chain CDR2 of SEQ ID NO:498, and the light chain CDR3 of SEQ ID NO:499; The heavy chain CDR1 of SEQ ID NO:511, the heavy chain CDR2 of SEQ ID NO:512, the heavy chain CDR3 of SEQ ID NO:513, the light chain CDR1 of SEQ ID NO:514, the light chain CDR2 of SEQ ID NO:515, and the light chain CDR3 of SEQ ID NO:516; The heavy chain CDR1 of SEQ ID NO:528, the heavy chain CDR2 of SEQ ID NO:529, the heavy chain CDR3 of SEQ ID NO:530, the light chain CDR1 of SEQ ID NO:531, the light chain CDR2 of SEQ ID NO:532, and the light chain CDR3 of SEQ ID NO:533; The heavy chain CDR1 of SEQ ID NO:545, the heavy chain CDR2 of SEQ ID NO:546, the heavy chain CDR3 of SEQ ID NO:547, the light chain CDR1 of SEQ ID NO:548, the light chain CDR2 of SEQ ID NO:549, and the light chain CDR3 of SEQ ID NO:550; The heavy chain CDR1 of SEQ ID NO:562, the heavy chain CDR2 of SEQ ID NO:563, the heavy chain CDR3 of SEQ ID NO:564, the light chain CDR1 of SEQ ID NO:565, the light chain CDR2 of SEQ ID NO:566, and the light chain CDR3 of SEQ ID NO:567; The heavy chain CDR1 of SEQ ID NO:579, the heavy chain CDR2 of SEQ ID NO:580, the heavy chain CDR3 of SEQ ID NO:581, the light chain CDR1 of SEQ ID NO:582, the light chain CDR2 of SEQ ID NO:583, and the light chain CDR3 of SEQ ID NO:584; The heavy chain CDR1 of SEQ ID NO:596, the heavy chain CDR2 of SEQ ID NO:597, the heavy chain CDR3 of SEQ ID NO:598, the light chain CDR1 of SEQ ID NO:599, the light chain CDR2 of SEQ ID NO:600, and the light chain CDR3 of SEQ ID NO:601; The heavy chain CDR1 of SEQ ID NO:613, the heavy chain CDR2 of SEQ ID NO:614, the heavy chain CDR3 of SEQ ID NO:615, the light chain CDR1 of SEQ ID NO:616, the light chain CDR2 of SEQ ID NO:617, and the light chain CDR3 of SEQ ID NO:618; The heavy chain CDR1 of SEQ ID NO:630, the heavy chain CDR2 of SEQ ID NO:631, the heavy chain CDR3 of SEQ ID NO:632, the light chain CDR1 of SEQ ID NO:633, the light chain CDR2 of SEQ ID NO:634, and the light chain CDR3 of SEQ ID NO:635; The heavy chain CDR1 of SEQ ID NO:647, the heavy chain CDR2 of SEQ ID NO:648, the heavy chain CDR3 of SEQ ID NO:649, the light chain CDR1 of SEQ ID NO:650, the light chain CDR2 of SEQ ID NO:651, and the light chain CDR3 of SEQ ID NO:652; The heavy chain CDR1 of SEQ ID NO:664, the heavy chain CDR2 of SEQ ID NO:665, the heavy chain CDR3 of SEQ ID NO:666, the light chain CDR1 of SEQ ID NO:667, the light chain CDR2 of SEQ ID NO:668, and the light chain CDR3 of SEQ ID NO:669; The heavy chain CDR1 of SEQ ID NO:681, the heavy chain CDR2 of SEQ ID NO:682, the heavy chain CDR3 of SEQ ID NO:683, the light chain CDR1 of SEQ ID NO:684, the light chain CDR2 of SEQ ID NO:685, and the light chain CDR3 of SEQ ID NO:686; The heavy chain CDR1 of SEQ ID NO:698, the heavy chain CDR2 of SEQ ID NO:699, the heavy chain CDR3 of SEQ ID NO:700, the light chain CDR1 of SEQ ID NO:701, the light chain CDR2 of SEQ ID NO:702, and the light chain CDR3 of SEQ ID NO:703; The heavy chain CDR1 of SEQ ID NO:715, the heavy chain CDR2 of SEQ ID NO:716, the heavy chain CDR3 of SEQ ID NO:717, the light chain CDR1 of SEQ ID NO:718, the light chain CDR2 of SEQ ID NO:719, and the light chain CDR3 of SEQ ID NO:720; The heavy chain CDR1 of SEQ ID NO:732, the heavy chain CDR2 of SEQ ID NO:733, the heavy chain CDR3 of SEQ ID NO:734, the light chain CDR1 of SEQ ID NO:735, the light chain CDR2 of SEQ ID NO:736, and the light chain CDR3 of SEQ ID NO:737; The heavy chain CDR1 of SEQ ID NO:749, the heavy chain CDR2 of SEQ ID NO:750, the heavy chain CDR3 of SEQ ID NO:751, the light chain CDR1 of SEQ ID NO:752, the light chain CDR2 of SEQ ID NO:753, and the light chain CDR3 of SEQ ID NO:754; The heavy chain CDR1 of SEQ ID NO:766, the heavy chain CDR2 of SEQ ID NO:767, the heavy chain CDR3 of SEQ ID NO:768, the light chain CDR1 of SEQ ID NO:769, the light chain CDR2 of SEQ ID NO:770, and the light chain CDR3 of SEQ ID NO:771; The heavy chain CDR1 of SEQ ID NO:783, the heavy chain CDR2 of SEQ ID NO:784, the heavy chain CDR3 of SEQ ID NO:785, the light chain CDR1 of SEQ ID NO:786, the light chain CDR2 of SEQ ID NO:787, and the light chain CDR3 of SEQ ID NO:788; The heavy chain CDR1 of SEQ ID NO:800, the heavy chain CDR2 of SEQ ID NO:801, the heavy chain CDR3 of SEQ ID NO:802, the light chain CDR1 of SEQ ID NO:803, the light chain CDR2 of SEQ ID NO:804, and the light chain CDR3 of SEQ ID NO:805; The heavy chain CDR1 of SEQ ID NO:817, the heavy chain CDR2 of SEQ ID NO:818, the heavy chain CDR3 of SEQ ID NO:819, the light chain CDR1 of SEQ ID NO:820, the light chain CDR2 of SEQ ID NO:821, and the light chain CDR3 of SEQ ID NO:822; The heavy chain CDR1 of SEQ ID NO:834, the heavy chain CDR2 of SEQ ID NO:835, the heavy chain CDR3 of SEQ ID NO:836, the light chain CDR1 of SEQ ID NO:837, the light chain CDR2 of SEQ ID NO:838, and the light chain CDR3 of SEQ ID NO:839; The heavy chain CDR1 of SEQ ID NO:851; the heavy chain CDR2 of SEQ ID NO:852; the heavy chain CDR3 of SEQ ID NO:853; the light chain CDR1 of SEQ ID NO:854; the light chain CDR2 of SEQ ID NO:855; and the light chain CDR3 of SEQ ID NO:856; The heavy chain CDR1 of SEQ ID NO:868, the heavy chain CDR2 of SEQ ID NO:869, the heavy chain CDR3 of SEQ ID NO:870, the light chain CDR1 of SEQ ID NO:871, the light chain CDR2 of SEQ ID NO:872, and the light chain CDR3 of SEQ ID NO:873; The heavy chain CDR1 of SEQ ID NO:885, the heavy chain CDR2 of SEQ ID NO:886, the heavy chain CDR3 of SEQ ID NO:887, the light chain CDR1 of SEQ ID NO:888, the light chain CDR2 of SEQ ID NO:889, and the light chain CDR3 of SEQ ID NO:890; The heavy chain CDR1 of SEQ ID NO:902, the heavy chain CDR2 of SEQ ID NO:903, the heavy chain CDR3 of SEQ ID NO:904, the light chain CDR1 of SEQ ID NO:905, the light chain CDR2 of SEQ ID NO:906, and the light chain CDR3 of SEQ ID NO:907; The heavy chain CDR1 of SEQ ID NO:919; the heavy chain CDR2 of SEQ ID NO:920; and the heavy chain CDR3 of SEQ ID NO:921; the light chain CDR1 of SEQ ID NO:922; the light chain CDR2 of SEQ ID NO:923; and the light chain CDR3 of SEQ ID NO:924; The heavy chain CDR1 of SEQ ID NO:936, the heavy chain CDR2 of SEQ ID NO:937, the heavy chain CDR3 of SEQ ID NO:938, the light chain CDR1 of SEQ ID NO:939, the light chain CDR2 of SEQ ID NO:940, and the light chain CDR3 of SEQ ID NO:941; The heavy chain CDR1 of SEQ ID NO:953, the heavy chain CDR2 of SEQ ID NO:954, the heavy chain CDR3 of SEQ ID NO:955, the light chain CDR1 of SEQ ID NO:956, the light chain CDR2 of SEQ ID NO:957, and the light chain CDR3 of SEQ ID NO:958; The heavy chain CDR1 of SEQ ID NO:970, the heavy chain CDR2 of SEQ ID NO:971, the heavy chain CDR3 of SEQ ID NO:972, the light chain CDR1 of SEQ ID NO:973, the light chain CDR2 of SEQ ID NO:974, and the light chain CDR3 of SEQ ID NO:975; The heavy chain CDR1 of SEQ ID NO:987, the heavy chain CDR2 of SEQ ID NO:988, the heavy chain CDR3 of SEQ ID NO:989, the light chain CDR1 of SEQ ID NO:990, the light chain CDR2 of SEQ ID NO:991, and the light chain CDR3 of SEQ ID NO:992; The heavy chain CDR1 of SEQ ID NO: 1004, the heavy chain CDR2 of SEQ ID NO: 1005, the heavy chain CDR3 of SEQ ID NO: 1006, the light chain CDR1 of SEQ ID NO: 1007, the light chain CDR2 of SEQ ID NO: 1008, and the light chain CDR3 of SEQ ID NO: 1009; The heavy chain CDR1 of SEQ ID NO: 1021, the heavy chain CDR2 of SEQ ID NO: 1022, the heavy chain CDR3 of SEQ ID NO: 1023, the light chain CDR1 of SEQ ID NO: 1024, the light chain CDR2 of SEQ ID NO: 1025, and the light chain CDR3 of SEQ ID NO: 1026; The heavy chain CDR1 of SEQ ID NO: 1062; the heavy chain CDR2 of SEQ ID NO: 1064; and the heavy chain CDR3 of SEQ ID NO: 1065; the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068; or The heavy chain CDR1 of SEQ ID NO:1063; the heavy chain CDR2 of SEQ ID NO:1064; the heavy chain CDR3 of SEQ ID NO:1065; the light chain CDR1 of SEQ ID NO:1066; the light chain CDR2 of SEQ ID NO:1067; and the light chain CDR3 of SEQ ID NO:1068.

9. The antibody or fragment thereof according to claim 8, wherein the antibody has: The heavy chain CDR1 of SEQ ID NO: 1; the heavy chain CDR2 of SEQ ID NO: 2; and the heavy chain CDR3 of SEQ ID NO: 3, the light chain CDR1 of SEQ ID NO: 4; the light chain CDR2 of SEQ ID NO: 5; and the light chain CDR3 of SEQ ID NO: 6; The heavy chain CDR1 of SEQ ID NO:35; the heavy chain CDR2 of SEQ ID NO:36; and the heavy chain CDR3 of SEQ ID NO:37, the light chain CDR1 of SEQ ID NO:38; the light chain CDR2 of SEQ ID NO:39; and the light chain CDR3 of SEQ ID NO:40; The heavy chain CDR1 of SEQ ID NO: 120; the heavy chain CDR2 of SEQ ID NO: 121; and the heavy chain CDR3 of SEQ ID NO: 122, the light chain CDR1 of SEQ ID NO: 123; the light chain CDR2 of SEQ ID NO: 124; and the light chain CDR3 of SEQ ID NO: 125; The heavy chain CDR1 of SEQ ID NO:222; the heavy chain CDR2 of SEQ ID NO:223; and the heavy chain CDR3 of SEQ ID NO:224, the light chain CDR1 of SEQ ID NO:225; the light chain CDR2 of SEQ ID NO:226; and the light chain CDR3 of SEQ ID NO:227; The heavy chain CDR1 of SEQ ID NO:239; the heavy chain CDR2 of SEQ ID NO:240; and the heavy chain CDR3 of SEQ ID NO:241, the light chain CDR1 of SEQ ID NO:242; the light chain CDR2 of SEQ ID NO:243; and the light chain CDR3 of SEQ ID NO:244; The heavy chain CDR1 of SEQ ID NO:290; the heavy chain CDR2 of SEQ ID NO:291; and the heavy chain CDR3 of SEQ ID NO:292, the light chain CDR1 of SEQ ID NO:293; the light chain CDR2 of SEQ ID NO:294; and the light chain CDR3 of SEQ ID NO:295; The heavy chain CDR1 of SEQ ID NO:324; the heavy chain CDR2 of SEQ ID NO:325; and the heavy chain CDR3 of SEQ ID NO:326, the light chain CDR1 of SEQ ID NO:327; the light chain CDR2 of SEQ ID NO:328; and the light chain CDR3 of SEQ ID NO:329; The heavy chain CDR1 of SEQ ID NO:341; the heavy chain CDR2 of SEQ ID NO:342; and the heavy chain CDR3 of SEQ ID NO:343, the light chain CDR1 of SEQ ID NO:344; the light chain CDR2 of SEQ ID NO:345; and the light chain CDR3 of SEQ ID NO:346; The heavy chain CDR1 of SEQ ID NO:851; the heavy chain CDR2 of SEQ ID NO:852; and the heavy chain CDR3 of SEQ ID NO:853, the light chain CDR1 of SEQ ID NO:854; the light chain CDR2 of SEQ ID NO:855; and the light chain CDR3 of SEQ ID NO:856; The heavy chain CDR1 of SEQ ID NO:919; the heavy chain CDR2 of SEQ ID NO:920; and the heavy chain CDR3 of SEQ ID NO:921, the light chain CDR1 of SEQ ID NO:922; the light chain CDR2 of SEQ ID NO:923; and the light chain CDR3 of SEQ ID NO:924; The heavy chain CDR1 of SEQ ID NO: 1062; the heavy chain CDR2 of SEQ ID NO: 1064; and the heavy chain CDR3 of SEQ ID NO: 1065, the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068; or The heavy chain CDR1 of SEQ ID NO: 1063; the heavy chain CDR2 of SEQ ID NO: 1064; and the heavy chain CDR3 of SEQ ID NO: 1065, the light chain CDR1 of SEQ ID NO: 1066; the light chain CDR2 of SEQ ID NO: 1067; and the light chain CDR3 of SEQ ID NO: 1068.

10. A kit for treating cancer, comprising: An oncolytic virus comprising nucleotides encoding protein A56 or a fragment thereof; wherein the protein A56 or a fragment thereof comprises a polypeptide having at least 90% sequence homology to the amino acid sequence shown in SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079, and the protein A56 or a fragment thereof is specifically bound by the antibody or a fragment thereof according to claim 8; and The antibody or fragment thereof according to claim 8.

11. An oncolytic virus comprising a nucleotide encoding protein A56 or a fragment thereof; and use of the antibody or a fragment thereof as claimed in claim 8 for the preparation of a drug for treating cancer; wherein the protein A56 or a fragment thereof comprises a polypeptide having at least 90% sequence homology to the amino acid sequence shown in SEQ ID NO: 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1073, 1075, 1077 or 1079, and the protein A56 or a fragment thereof is specifically bound by the antibody or fragment thereof according to claim 8; Furthermore, the cancer is selected from the group consisting of lung cancer, colorectal cancer, breast cancer, uterine cancer and prostate cancer.

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