Antibody specifically binding to lipocortin 1 and use thereof

Abstract

The present invention relates to a molecule specifically binding to lipocortin 1 in primates and rodents, particularly a monoclonal antibody or an antigen-binding fragment thereof. An anti-lipocotin 1 antibody, according to the present invention, exhibited concentration-dependent specific binding affinity to recombinant and intracellular lipocotin 1 proteins. In addition, the antibody of the present invention exhibited cell permeability at a cellular level and exhibited drug efficacy at an animal level. Therefore, a monoclonal anti-lipocotin 1 antibody, according to the present invention, can be used for the treatment of various diseases associated with lipocotin 1.

Description

Antibody specifically binding to lipocotin 1 and uses thereof

[0001] The present invention relates to an antibody or an antigen-binding fragment thereof that specifically binds to lipocortin 1 of primates and rodents, and to the use thereof.

[0002] To date, 12 families of annexins (annexin A) have been discovered in mammals and are also known as lipocortin, calpactin, and endonexin. Annexins bind to calcium and phospholipids and possess a unique domain consisting of a sequence of about 70 amino acids repeated 4 or 8 times, containing a 'KGxGT' motif called the endonexin-fold at the C-terminus. All previously published annexin family proteins consist of the aforementioned conserved domain and N-terminal regions of varying lengths and order.

[0003] The aforementioned annexin proteins are known to be involved in various biological phenomena, such as bone structure formation, membrane trafficking, membrane permeability channel activation, inhibition of phospholipase A2, inhibition of coagulation, transmission of mitogen signals, and regulation of cell-matrix interactions. In addition, a deficiency of annexin proteins can cause various malformations in the human body and is known to be involved in the development of autoimmune diseases or cancer.

[0004] Among annexin proteins, lipocortin 1 (annexin A1), which belongs to the annexin I group, is also found in cancer tissues. In particular, lipocortin 1 is known to be overexpressed in cancer cells, as well as having increased expression in surrounding tissues and blood vessels.

[0005] In addition, lipocotin 1 blocks both innate and adaptive immune responses simultaneously by mediating the inflammatory regulatory response of glucocorticoids. Lipocotin 1 regulates homeostasis in various cells involved in innate and adaptive immunity, including neutrophils, macrophages, and endothelial cells. For example, lipocotin 1 regulates homeostasis in neutrophils and macrophages of the innate immune system and regulates T cell receptor signaling in T cells of the adaptive immune system.

[0006] Due to the biological functions of lipocotin 1, it is attracting attention as a target protein for the development of anticancer agents, drug delivery systems, anti-inflammatory agents, and immunomodulators, and there is a demand for the development of effective and potent targeted drugs that can utilize it (Korean Patent Publication No. 10-2015-0029457).

[0007] Accordingly, the inventors of the present invention investigated a method to effectively inhibit the activity of lipocotin 1, developed an antibody that specifically binds to lipocotin 1, and completed the present invention by confirming the anticancer activity of the said antibody.

[0008] To achieve the above objective, one aspect of the present invention comprises: (a) a heavy chain variable region (VH) comprising: i) a heavy chain complementarity determining region 1 (HCDR1) represented by the amino acid sequence of SEQ ID NO. 210; ii) a heavy chain complementarity determining region 2 (HCDR2) represented by the amino acid sequence of SEQ ID NO. 211; and iii) a heavy chain complementarity determining region 3 (HCDR3) represented by the amino acid sequence of SEQ ID NO. 212; and (b) iv) a light chain complementarity determining region 1 (LCDR1) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 53 to 72, SEQ ID NO. 225, and SEQ ID NO. 226; and v) a light chain complementarity determining region 2 (LCDR2) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 73 to 84 and SEQ ID NO. 227; and vi) provide an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region (VL) comprising a light chain complementarity determining region 3 (LCDR3) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 85 to 108, SEQ ID NOs 217, and SEQ ID NOs 228 to 230. Herein, in the GX1X2X3X4X5X6X7 amino acid sequence of SEQ ID NO. 210, X1 is F, I, Y, or N; X2 is T, S, N, or I; X3 is F, L, or T; X4 is G, D, N, S, or T; X5 is D, M, E, G, T, or S; X6 is Y, H, F, or L; and X7 is A, P, G, S, Y, or W; and IRX8X9X of SEQ ID NO. 211 10 X 11 GGTX 12 X8 of the amino acid sequence is S, T, D, or P, X9 is K, A, or R, and X 10 is A, S, T, R, W, Q or L, and X 11 is Y, F, I, V, L or T, and X 12 is T or P; X of sequence number 21213 RDGNYYDSRX 14 YYYDTFDX 15 X of the amino acid sequence 13 is A or V, and X 14 is G or D, and X 15 is L or M.

[0009] Another aspect of the present invention comprises a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 1, HCDR2 represented by the amino acid sequence of SEQ ID NO. 23, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 42, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 85; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 2, HCDR2 represented by the amino acid sequence of SEQ ID NO. 24, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 43, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 85; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 3, HCDR2 represented by the amino acid sequence of SEQ ID NO. 25, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 44, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 74, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 86; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 4, HCDR2 represented by the amino acid sequence of SEQ ID NO. 26, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 45, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 87;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 5, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 54, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 88; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 6, HCDR2 represented by the amino acid sequence of SEQ ID NO. 28, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 47, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 55, LCDR2 represented by the amino acid sequence of SEQ ID NO. 76, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 89; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 7, HCDR2 represented by the amino acid sequence of SEQ ID NO. 29, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 48, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 56, LCDR2 represented by the amino acid sequence of SEQ ID NO. 77, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 90; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 49, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 57, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 88;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 79, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 91; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 9, HCDR2 represented by the amino acid sequence of SEQ ID NO. 31, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 59, LCDR2 represented by the amino acid sequence of SEQ ID NO. 80, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 92; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 10, HCDR2 represented by the amino acid sequence of SEQ ID NO. 32, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 50, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 60, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 93; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 11, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 94;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 33, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 51, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 61, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 95; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 96; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 63, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 98;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 64, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 99; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 13, HCDR2 represented by the amino acid sequence of SEQ ID NO. 34, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 65, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 14, HCDR2 represented by the amino acid sequence of SEQ ID NO. 35, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 98; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 15, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 66, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 100;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 67, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 101; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 68, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 102; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 103; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 16, HCDR2 represented by the amino acid sequence of SEQ ID NO. 36, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 69, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 104;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 17, HCDR2 represented by the amino acid sequence of SEQ ID NO. 37, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 70, LCDR2 represented by the amino acid sequence of SEQ ID NO. 84, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 105; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 18, HCDR2 represented by the amino acid sequence of SEQ ID NO. 38, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 106; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 19, HCDR2 represented by the amino acid sequence of SEQ ID NO. 39, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 71, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 107; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 20, HCDR2 represented by the amino acid sequence of SEQ ID NO. 40, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 108;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 21, HCDR2 represented by the amino acid sequence of SEQ ID NO. 41, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 108; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 217; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97;A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 215, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; or a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 216, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 222, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228; a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 223, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 226, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 229;Provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 224, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 230; a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228.

[0010] Another aspect of the present invention provides a polynucleotide encoding the antibody or its antigen-binding fragment, a vector loaded with the polynucleotide, and a cell transformed with the vector.

[0011] Another aspect of the present invention provides a method for producing an antibody or an antigen-binding fragment that specifically binds to lipocotin 1, comprising the steps of: ii) culturing the transformed cells; and ii) obtaining an antibody or an antigen-binding fragment thereof from a culture medium of the cells.

[0012] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising the antibody or its antigen-binding fragment as an active ingredient.

[0013] Another aspect of the present invention provides an antibody-drug conjugate comprising the antibody or an antigen-binding fragment thereof; and an anticancer agent bound thereto.

[0014] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising the antibody-drug conjugate as an active ingredient.

[0015] Another aspect of the present invention provides the use of the antibody or the antibody-drug conjugate for the prevention or treatment of cancer.

[0016] Another aspect of the present invention provides a method for preventing or treating cancer, comprising the step of administering the antibody or the antibody-drug conjugate to an individual.

[0017] The anti-lipocotin 1 antibody according to the present invention exhibited concentration-dependent specific binding affinity to recombinant and intracellular lipocotin 1 proteins. In addition, the antibody exhibited cell permeability and drug transport capacity at the cellular and animal levels. Therefore, the monoclonal anti-lipocotin 1 antibody according to the present invention can be used to treat various diseases associated with lipocotin 1.

[0018] Figure 1 shows the results of confirming the antigen protein purification product through SDS-PAGE under reduced conditions. Figure 1a shows the results of analyzing the human lipocotin 1 antigen protein (LPC-1-his), and Figure 1b shows the results of comparative analysis of the human lipocotin 1 antigen protein (LPC-1-his), mouse lipocotin 1 antigen protein (mLPC-1-his), and monkey lipocotin 1 antigen protein (moLPC-1-his). Figure 1c is a diagram showing the results of purifying and analyzing the lipocotin 1 mutant antigen according to the present invention.

[0019] Figures 2a, 2b, 3a, and 3b are the results of confirming the specific binding affinity of scFv-type heat clones (100 types) obtained through biopanning to lipocotin 1 using Direct ELISA.

[0020] Figure 4 shows the results of analyzing purified IgG form HEAT antibodies. Figures 4a to 4d show the results of analyzing 97 types of purified HEAT antibody products, and Figures 4e and 4f show the results of analyzing 8 types of purified antibody products with mature affinities using SDS-PAGE under reduced conditions.

[0021] Figures 5a and 5b show the results of confirming the binding affinity of 94 types of anti-lipocotin 1 antibodies to human antigens (LPC-1-his with 26 amino acids deleted from the N-terminus or full-length human lipocotin 1 protein (AA-1)) via ELISA.

[0022] Figure 6 shows the results of confirming the binding affinity of each anti-lipocotin 1 antibody to the antigen at different concentrations using ELISA. Figure 6a shows the results of confirming the binding affinity of the IgG form of the HEAT antibody to the human lipocotin 1 antigen (LPC-1-his), and Figure 6b shows the results of confirming the binding affinity of the HEAT antibody to the mouse lipocotin 1 antigen (mLPC-1-his). In addition, Figure 6c shows the results of confirming the binding power of four types of mature affinity antibodies to each antigen (LPC-1-his, mLPC-1-his, moLPC-1-his) (0.1 μg / mL), Figure 6d shows the results of confirming the binding power of four types of mature affinity antibodies to the full-length antigen (0.1 μg / mL AA-1) and family members (0.5 μg / mL AA-2, 0.5 μg / mL AA-5), and Figure 6e shows the results of confirming the concentration-dependent binding power of four types of mature affinity antibodies to the human lipocotinic 1 antigen (LPC-1-his).

[0023] Figure 7 shows the results of confirming the binding affinity of an anti-lipocotin 1 antibody to lipocotin 1 expressed in human cell lines through flow cytometry. Figure 7a shows the results of confirming the binding affinity of a HEAT antibody to HUVEC cells or A549 cells (human cell lines) expressing lipocotin 1, and Figure 7b shows the results of confirming the binding affinity of a HEAT antibody to NIH-3T3 cells (mouse cell lines) expressing lipocotin 1.

[0024] Figure 8 shows the results of confirming the binding affinity of anti-lipocotin 1 antibodies at different concentrations in human cell lines expressing lipocotin 1 through flow cytometry. Figures 8a and 8b show the results of confirming the binding affinity of the hit antibody in HUVEC cell lines or A549 cell lines, respectively. In addition, Figure 8c shows the results of confirming the binding affinity of the hit antibody to NIH-3T3 cell lines. Figures 8d and 8e show the results of confirming the binding affinity of four antibodies with mature affinity in A549 cell lines (fixation conditions) or HUVEC cell lines, respectively.

[0025] Figure 9 shows the results of confirming the binding affinity of lipocotin 1-specific binding antibodies to the antigen (LPC-1-his) using the BLI method. Figure 9a shows the results of confirming the binding affinity of the hit antibody (TRO22O6E) to the antigen (LPC-1-his), and Figure 9b shows the results of confirming the binding affinity of the hit antibody (TRO22O9G) to the antigen (LPC-1-his). In addition, Figure 9c shows the results of confirming the binding affinity of the hit antibody (TRO22H2I) to the antigen (LPC-1-his), and Figure 9d shows the results of confirming the binding affinity of the affinity-matured antibody (H2I23DID) to the antigen (LPC-1-his). Figure 9e shows the results of confirming the binding affinity of the hit antibody (H2I24095) to the antigen (LPC-1-his), and Figure 9f shows the results of confirming the binding affinity of the hit antibody (H2I24109) to the antigen (LPC-1-his). Figure 9g shows the results of confirming the binding affinity of the hit antibody (H2I24128) to the antigen (LPC-1-his), and Figure 9h shows the results of confirming the binding affinity of the hit antibody (H2I24132) to the antigen (LPC-1-his).

[0026] Figure 10 shows the results of confirming the binding affinity of lipocotin 1-specific binding antibodies to the antigen (LPC-1-his) using the SPR method. Figure 10a shows the results of confirming the binding affinity of the hit antibody (TRO22H2I) to the antigen (LPC-1-his), and Figure 10b shows the results of confirming the binding affinity of the hit antibody (H2I24095) to the antigen (LPC-1-his). Figure 10c shows the results of confirming the binding affinity of the hit antibody (H2I24109) to the antigen (LPC-1-his), Figure 10d shows the results of confirming the binding affinity of the affinity-mature antibody (H2I24128) to the antigen (LPC-1-his), and Figure 10e shows the results of confirming the binding affinity of the affinity-mature antibody (H2I24132) to the antigen (LPC-1-his).

[0027] Figure 11 shows the results of confirming the binding affinity to mutant antigen proteins using ELISA to analyze the antigen (LPC-1-his) binding site of lipocotin 1-specific binding antibodies. Figure 11a shows the results of analyzing the binding affinity of TRO22H2I antibodies against chimeric protein antigens in which four domains of human lipocotin 1 protein (AA1) were each substituted with domains of lipocotin 2 protein (AA2). Figure 11b shows the results of analyzing the binding affinity of hit antibodies that specifically bind to lipocotin 1 protein (AA1) against site-specific mutations in domains 1 and 2 of human lipocotin 1 protein (AA1). Figure 11c shows the results of analyzing the binding affinity of each hit antibody against a single amino acid mutant antigen of human lipocotin 1 protein (AA1).

[0028] Figure 12 shows the results of confirming the extracellular transport efficiency of the HEAT antibody under co-culture conditions of HUVEC cell line and MCF-7 cell line.

[0029] Figure 13 shows the results confirming the antitumor efficacy of the anti-lipocotin 1 hit antibody in a JIMT-1 cell line xenograft mouse tumor model. Figures 13a and 13b show the results of measuring tumor volume and mouse body weight, respectively, after administering an antibody-drug conjugate in which MMAF (monomethyl auristatin F) was bound to the anti-lipocotin 1 hit antibody (TRO22H2I) in a JIMT-1 cell line xenograft mouse tumor model.

[0030] The present invention provides an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof.

[0031] Anti-lipocotin 1 antibody

[0032] The term "lipocortin 1" as used herein is also known as annexin A1, calpactin II, chromobindin-9, phospholipase A2 inhibitory protein, and p35. In general, lipocortin 1 is known to be involved in intercellular signaling and its function is regulated by calcium (Ca) concentration. Additionally, lipocortin 1 is considered a factor affecting the growth and metastasis of cancer cells, and overexpression of lipocortin 1 in various cancers has been reported. In the present invention, the lipocortin 1 may be used interchangeably with "annexin A1".

[0033] In the present invention, lipocotin 1 may be included without limitation if it is of mammalian origin, including primates such as humans and monkeys, and rodents such as rats and mice.

[0034] In addition, the above lipocotin 1 protein may include both natural and variant lipocotin 1 proteins, but is not limited thereto. The above natural lipocotin 1 protein generally refers to a polypeptide containing the amino acid sequence of the natural lipocotin 1 protein. The amino acid sequence and polynucleotide sequence for the above lipocotin 1 can be obtained from known databases such as the National Institutes of Health (NCBI) GenBank, and preferably may include the amino acid sequence of SEQ ID NO. 206. In addition, as long as it has the same activity as the above protein or the same gene location encoding the above lipocotin 1 on the chromosome, it may be composed of a sequence in which one or several amino acids of the above protein are added, deleted, or substituted. Specifically, lipocotin 1 may include or be composed of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity with the amino acid sequence of SEQ ID NO. 206.

[0035] In the present invention, the lipocotin 1 protein variant refers to a protein in which a portion of the N-terminus is deleted. Specifically, the lipocotin 1 protein variant may be in the form of an amino acid in which the 2nd to 26th amino acids from the N-terminus of a protein having the amino acid sequence of SEQ ID NO. 206 are consecutively deleted. Specifically, it may be one that includes the amino acid sequence of SEQ ID NO. 213.

[0036] In addition, it may be composed of a sequence having the same activity as the above variant or having the same gene location encoding the above lipocotin 1 on the chromosome, in which one or several amino acids are added, deleted, or substituted. Specifically, it may include or be composed of an amino acid sequence having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity with the amino acid sequence of SEQ ID NO. 213.

[0037] The term "anti-lipocotin 1 antibody" as used in this specification refers to an antibody capable of binding to lipocotin 1, and may be used interchangeably with "antibody specific to lipocotin 1" or "antibody specifically binding to lipocotin 1" in this specification. In particular, the anti-lipocotin 1 antibody can specifically bind not only to natural lipocotin 1 (SEQ No. 206) but also to a variant of natural lipocotin 1 in which the N-terminus is deleted (SEQ No. 213).

[0038] As used in this specification, the term "antibody" refers to an immunoglobulin (Ig) molecule that reacts immunologically with a specific antigen, meaning a protein molecule that acts as a receptor specifically recognizing the antigen, and is a concept that encompasses both whole antibodies and antibody fragments.

[0039] Specifically, the antibody or its antigen-binding fragment may include, but is not limited to, monoclonal antibodies, polyclonal antibodies, single domain antibodies, single chain antibodies, multispecific antibodies, human antibodies, humanized antibodies, chimeric antibodies, intrabody, Fv, scFv, Fv (di-scFv) connected by disulfide bonds, Fab fragments, F(ab')2 fragments, and any of the epitope-binding fragments.

[0040] In the present invention, the antibody comprises a heavy chain and a light chain, and the heavy chain and the light chain may each comprise a constant region and a variable region.

[0041] The term “heavy chain” as used in this specification means a heavy chain of full length including a variable region (VH) sufficient to confer specificity to an antigen and three constant regions CH1, CH2 and CH3, and all fragments thereof.

[0042] As used herein, the term "light chain" means including both the full-length light chain and fragments thereof, which contain a variable region (VL) and a constant region (CL) sufficient to impart specificity to an antigen.

[0043] The light and heavy chain variable regions of the above antibody include three hypervariable regions and four framework regions (FR) called complementarity determining regions (CDRs). The CDRs primarily serve to bind to the epitopes of the antigen. The CDRs of each chain are typically named sequentially starting from the N-terminus as CDR1, CDR2, and CDR3, and are identified by the chain in which a specific CDR is located.

[0044] Accordingly, one aspect of the present invention comprises: (a) a heavy chain variable region (VH) comprising i) a heavy chain complementarity determining region 1 (HCDR1) represented by the amino acid sequence of SEQ ID NO. 210; ii) a heavy chain complementarity determining region 2 (HCDR2) represented by the amino acid sequence of SEQ ID NO. 211; and iii) a heavy chain complementarity determining region 3 (HCDR3) represented by the amino acid sequence of SEQ ID NO. 212; and (b) iv) a light chain complementarity determining region 1 (LCDR1) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 53 to 72, SEQ ID NO. 225, and SEQ ID NO. 226; and v) a light chain complementarity determining region 2 (LCDR2) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 73 to 84 and SEQ ID NO. 227; and vi) provide an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region (VL) comprising a light chain complementarity determining region 3 (LCDR3) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 85 to 108, SEQ ID NOs 217, and SEQ ID NOs 228 to 230. Herein, in the GX1X2X3X4X5X6X7 amino acid sequence of SEQ ID NO. 210, X1 is F, I, Y, or N; X2 is T, S, N, or I; X3 is F, L, or T; X4 is G, D, N, S, or T; X5 is D, M, E, G, T, or S; X6 is Y, H, F, or L; and X7 is A, P, G, S, Y, or W; and IRX8X9X of SEQ ID NO. 211 10 X 11 GGTX 12 X8 of the amino acid sequence is S, T, D, or P, X9 is K, A, or R, and X 10 is A, S, T, R, W, Q or L, and X 11 is Y, F, I, V, L or T, and X 12 is T or P; X of sequence number 212 13RDGNYYDSRX 14 YYYDTFDX 15 X of the amino acid sequence 13 is A or V, and X 14 is G or D, and X 15 is L or M.

[0045] Specifically, the heavy chain variable region (VH) may include an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof, comprising: i) a heavy chain complementarity determining region 1 (HCDR1) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 1 to 22; ii) a heavy chain complementarity determining region 2 (HCDR2) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 23 to 41, SEQ ID NOs 214 to 216, and SEQ ID NOs 222 to 224; and iii) a heavy chain complementarity determining region 3 (HCDR3) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 42 to 52.

[0046] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 1, HCDR2 represented by the amino acid sequence of SEQ ID NO. 23, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 42; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 85. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 109, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 139.

[0047] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 2, HCDR2 represented by the amino acid sequence of SEQ ID NO. 24, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 43; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 85. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 110, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 140.

[0048] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 3, HCDR2 represented by the amino acid sequence of SEQ ID NO. 25, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 44; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 74, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 86. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 111, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 141.

[0049] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 4, HCDR2 represented by the amino acid sequence of SEQ ID NO. 26, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 45; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 87. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 112, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 142.

[0050] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 5, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 54, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 88. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 113, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 143.

[0051] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 6, HCDR2 represented by the amino acid sequence of SEQ ID NO. 28, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 47; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 55, LCDR2 represented by the amino acid sequence of SEQ ID NO. 76, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 89. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 114, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 144.

[0052] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 7, HCDR2 represented by the amino acid sequence of SEQ ID NO. 29, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 48; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 56, LCDR2 represented by the amino acid sequence of SEQ ID NO. 77, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 90. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 115, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 145.

[0053] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 49; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 57, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 88. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 116, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 146.

[0054] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 79, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 91. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 117, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 147.

[0055] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 9, HCDR2 represented by the amino acid sequence of SEQ ID NO. 31, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 59, LCDR2 represented by the amino acid sequence of SEQ ID NO. 80, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 92. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 118, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 148.

[0056] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 10, HCDR2 represented by the amino acid sequence of SEQ ID NO. 32, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 50; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 60, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 93. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 119, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 149.

[0057] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 11, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 94. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 120, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 150.

[0058] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 33, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 51; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 61, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 95. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 121, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 151.

[0059] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 96. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 122, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 152.

[0060] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 63, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 123, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 153.

[0061] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 98. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 124, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 154.

[0062] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 64, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 99. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 125, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 155.

[0063] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 13, HCDR2 represented by the amino acid sequence of SEQ ID NO. 34, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 65, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 126, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 156.

[0064] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 14, HCDR2 represented by the amino acid sequence of SEQ ID NO. 35, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 98. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 127, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 157.

[0065] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 15, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 66, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 100. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 128, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 158.

[0066] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 67, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 101. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 129, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 159.

[0067] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 68, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 102. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 130, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 160.

[0068] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 103. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 131, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 161.

[0069] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 16, HCDR2 represented by the amino acid sequence of SEQ ID NO. 36, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 69, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 104. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 132, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 162.

[0070] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 17, HCDR2 represented by the amino acid sequence of SEQ ID NO. 37, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 70, LCDR2 represented by the amino acid sequence of SEQ ID NO. 84, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 105. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 133, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 163.

[0071] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 18, HCDR2 represented by the amino acid sequence of SEQ ID NO. 38, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 106. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 134, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 164.

[0072] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 19, HCDR2 represented by the amino acid sequence of SEQ ID NO. 39, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 71, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 107. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 135, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 165.

[0073] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 20, HCDR2 represented by the amino acid sequence of SEQ ID NO. 40, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 108. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 136, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 166.

[0074] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 21, HCDR2 represented by the amino acid sequence of SEQ ID NO. 41, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 108. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 137, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 167.

[0075] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 138, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 168.

[0076] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 217. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 138, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 221.

[0077] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 218, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 168.

[0078] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 215, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 219, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 168.

[0079] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 216, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 220, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 168.

[0080] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 222, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 231, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 235.

[0081] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 223, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 226, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 229. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 232, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 236.

[0082] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 224, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 230. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 233, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 237.

[0083] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228. wherein the heavy chain variable region may comprise the amino acid sequence of SEQ ID NO. 234, and the light chain variable region may comprise the amino acid sequence of SEQ ID NO. 238.

[0084] The heavy chain variable region of the above antibody may include or be composed of amino acid sequences having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity with each of the amino acid sequences of SEQ ID NOs 109 to 138, SEQ ID NOs 218 to 220, and SEQ ID NOs 231 to 234. In addition, the light chain variable region of the antibody may include or be composed of amino acid sequences having about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identity, or 100% identity with each of the amino acid sequences of SEQ ID NOs 139 to 168, SEQ ID NO 221, and SEQ ID NOs 235 to 238.

[0085] The heavy chain constant region (CH) of immunoglobulins exhibits different amino acid compositions and sequences, thereby possessing different types of antigenicity. Therefore, immunoglobulins can be classified into five categories and referred to as immunoglobulin isoforms, namely IgM, IgD, IgG, IgA, and IgE. The corresponding heavy chains are the μ chain, δ chain, γ chain, α chain, and ε chain, respectively. Furthermore, depending on the amino acid composition of the hinge region and the number and location of heavy chain disulfide bonds, the same type of Ig can be classified into different subtypes. For example, IgG can be classified into IgG1, IgG2, IgG3, and IgG4. Light chains can be classified as κ or λ chains depending on the different constant regions. Each of the five types of IgG can possess either a κ or λ chain. Here, immunoglobulin refers to a glycoprotein that acts as an antibody, synonymous with antibody.

[0086] When the antibody that specifically binds to lipocotin 1 of the present invention includes a constant region, it may include a constant region derived from IgG, IgA, IgD, IgE, IgM, or a partially mixed (hybrid) of these.

[0087] The term “hybrid” as used in this specification means that there are sequences corresponding to immunoglobulin heavy chain constant regions of two or more different origins within a single-chain immunoglobulin heavy chain constant region. For example, a hybrid of domains consisting of one to four domains selected from the group consisting of CH1, CH2, and CH3 of IgG, IgA, IgD, IgE, and IgM is possible.

[0088] In addition, if the antibody that specifically binds to the lipocotin 1 includes a light chain constant region (LC), the light chain constant region may be derived from a κ or λ light chain.

[0089] Polynucleotide encoding the anti-lipocotin 1 antibody

[0090] Another aspect of the present invention provides a polynucleotide encoding an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof. Specifically, the invention provides a polynucleotide encoding a heavy chain variable region comprising the respective amino acid sequences of SEQ ID NOs 109 to 138, SEQ ID NOs 218 to 220, and SEQ ID NOs 231 to 234; or a polynucleotide encoding a light chain variable region comprising the respective amino acid sequences of SEQ ID NOs 139 to 168, SEQ ID NOs 221, and SEQ ID NOs 235 to 238. The anti-lipocotin 1 antibody or the antigen-binding fragment thereof is the same as described above.

[0091] If the above polynucleotides encode the same polypeptide, one or more bases may be modified by substitution, deletion, insertion, or a combination thereof. When preparing polynucleotide sequences by chemical synthesis, synthesis methods widely known in the art may be used, such as the method described in the literature (Engels and Uhlmann, Angew Chem IntEd Engl., 37:73-127, 1988), as well as tryst, phosphate, phosphamidite, and H-phosphate methods, PCR and other autoprimer methods, and oligonucleotide synthesis methods on solid supports.

[0092] In addition, the polynucleotide may additionally include a signal sequence or a lead sequence.

[0093] As used herein, the term "signal sequence" refers to a nucleic acid encoding a signal peptide that directs the secretion of a target protein. The signal peptide is cleaved after translation in a host cell. Specifically, the signal sequence of the present invention is a nucleotide encoding an amino acid sequence that initiates the movement of a protein through the ER (endoplasmic reticulum) membrane.

[0094] The signal sequence is well known in the art for its characteristics and typically contains 16 to 30 amino acid residues, but may contain more or fewer amino acid residues. A typical signal peptide consists of three regions: a basic N-terminal region, a central hydrophobic region, and a more polar C-terminal region. The central hydrophobic region contains 4 to 12 hydrophobic residues that anchor the signal sequence through the membrane lipid bilayer while the immature polypeptide moves. After initiation, the signal sequence is cleaved within the lumen of the ER by cellular enzymes commonly known as signal peptidases. At this time, the signal sequence may be tPa (tissue plasminogen activation), HSV gDs (signal sequence of herpes simplex virus glycoprotein D), an IgG signal sequence, or a secretion signal sequence of growth hormone. Preferably, a secretion signal sequence used in higher eukaryotic cells, including mammals, may be used.

[0095] The signal sequences useful in the present invention may include antibody light chain signal sequences, e.g., antibody 14.18 (Gillies et al., J. Immunol. Meth 1989. 125:191-202), antibody heavy chain signal sequences, e.g., MOPC141 antibody heavy chain signal sequence (Sakano et al., Nature, 1980. 286: 676-683), and other signal sequences known in the art (e.g., see Watson et al., Nucleic Acid Research, 1984. 12:5145-5164).

[0096] As used herein, the term "lead sequence" refers to a region of messenger RNA (mRNA) upstream of the start codon that is not translated into protein.

[0097] Vectors containing polynucleotides

[0098] Another aspect of the present invention provides a vector comprising a polynucleotide encoding an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof. Specifically, the present invention provides a vector comprising: a polynucleotide encoding a heavy chain variable region comprising the respective amino acid sequences of SEQ ID NOs 109 to 138, SEQ ID NOs 218 to 220, and SEQ ID NOs 231 to 234; and a polynucleotide encoding a light chain variable region comprising the respective amino acid sequences of SEQ ID NOs 139 to 168, SEQ ID NOs 221, and SEQ ID NOs 235 to 238.

[0099] The above vector may be two vectors each containing a polynucleotide encoding the heavy chain and the light chain, or a bicistronic expression vector containing both of the polynucleotides.

[0100] As used herein, the term “vector” may be introduced into a host cell and recombinated and inserted into the host cell genome. Alternatively, the vector is understood as a nucleic acid means comprising a nucleotide sequence capable of spontaneously replicating as an episome. The vector comprises linear nucleic acids, plasmids, phagesmids, cosmids, RNA vectors, viral vectors, minichromosomes, and analogs thereof. Examples of viral vectors include, but are not limited to, retroviruses, adenoviruses, and adeno-associated viruses.

[0101] Specifically, the vector may be plasmid DNA, phage DNA, etc., and may be commercially developed plasmids (e.g., pUC18, pBAD, pIDTSAMRT-AMP, etc.), E. coli-derived plasmids (e.g., pYG601BR322, pBR325, pUC118, pUC119, etc.), Bacillus subtilis-derived plasmids (e.g., pUB110, pTP5, etc.), yeast-derived plasmids (e.g., YEp13, YEp24, YCp50, etc.), phage DNA (e.g., Charon4A, Charon21A, EMBL3, EMBL4, λgt10, λgt11, λZAP, etc.), animal virus vectors (e.g., retrovirus, adenovirus, vaccinia virus, etc.), insect virus vectors (e.g., baculovirus, etc.). Since the protein expression levels and modifications of the above vector vary depending on the host cell, it is desirable to select and use the host cell most suitable for the purpose.

[0102] In addition, the plasmid may include a selection marker such as an antibiotic resistance gene, and the host cell maintaining the plasmid may be cultured under selective conditions.

[0103] As used herein, the terms “gene expression” or “expression” of the target protein are understood to mean the transcription of a DNA sequence, the translation of an mRNA transcript, and the secretion of an antibody or its antigen-binding fragment. A useful expression vector may be RcCMV (Invitrogen) or a variant thereof. The expression vector may include a human CMV (cytomegalovirus) promoter to promote the continuous transcription of the target gene in mammalian cells and a bovine growth hormone polyadenylation signal sequence to increase the steady-state level of post-transcriptional RNA.

[0104] Transformed cells expressing anti-lipocotin 1 antibody

[0105] Another aspect of the present invention provides a transformed cell into which a vector comprising a polynucleotide encoding an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof has been introduced. The antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof is the same as described above.

[0106] As used herein, the term "transformed cell" refers to a prokaryotic cell and a eukaryotic cell into which a recombinant expression vector can be introduced. The transformed cell can be produced by introducing the vector into a host cell and transforming it. Additionally, an antibody or an antigen-binding fragment thereof that specifically binds to Lipocotin 1 of the present invention can be produced by expressing the polynucleotide contained in the vector.

[0107] The above transformation can be performed by various methods. As long as an antibody that specifically binds to Lipocotin 1 of the present invention or an antigen-binding fragment thereof can be produced, it is not particularly limited thereto. Specifically, the transformation method may include the CaCl2 precipitation method, the Hanahan method which increases efficiency by using a reducing agent called DMSO (dimethyl sulfoxide) in the CaCl2 precipitation method, electroporation, calcium phosphate precipitation method, protoplasmic fusion method, stirring method using silicon carbide fibers, Agrobacterium-mediated transformation method, PEG-mediated transformation method, dextran sulfate, lipofectamine, and drying / inhibition-mediated transformation method. Additionally, the target product may be delivered into the cell using viral particles by means of infection. Furthermore, the vector may be introduced into the host cell by means of gene bombardment, etc.

[0108] In addition, the host cell used for producing the above-mentioned transformed cell is not particularly limited thereto, as long as it is capable of producing the antibody of the present invention or its antigen-binding fragment. Specifically, the host cell may include, but is not limited to, prokaryotic cells, eukaryotic cells, mammalian, plant, insect, fungal, or cellular cells. As an example of the prokaryotic cell, Escherichia coli may be used. In addition, as an example of the eukaryotic cell, yeast may be used. Furthermore, as the mammalian cell, CHO cells, F2N cells, COS cells, BHK cells, Bowes melanoma cells, HeLa cells, 911 cells, AT1080 cells, A549 cells, SP2 / 0 cells, human lymphoblastoid, NSO cells, HT-1080 cells, PERC.6 cells, HEK293 cells, or HEK293T cells may be used, but is not limited thereto, and any cell known to those skilled in the art that can be used as a mammalian host cell may be used.

[0109] To optimize the therapeutic properties of the above anti-lipocotin 1 antibody or its antigen-binding fragment, or for other purposes, glycosylation-related genes possessed by the host cell can be manipulated through methods known to those skilled in the art to adjust the glycosylation pattern (e.g., sialic acid, fucosylation, glycosylation) of the antibody or its antigen-binding fragment that specifically binds to lipocotin 1.

[0110] Method for preparing anti-lipocotin 1 antibody

[0111] Another aspect of the present invention provides a method for preparing an antibody or an antigen-binding fragment thereof that specifically binds to the lipocotin 1. The antibody or antigen-binding fragment thereof that specifically binds to the lipocotin 1 is the same as described above.

[0112] A method for preparing an antibody or an antigen-binding fragment thereof that specifically binds to the above-mentioned lipocotin 1 may comprise: i) a step of culturing the above-mentioned transformed cells; and ii) a step of obtaining an anti-lipocotin 1 antibody or an antigen-binding fragment thereof from the cell culture. Herein, the transformed cells are as described above.

[0113] The method of culturing the above-mentioned transformed cells can be carried out using methods widely known in the art. The above-mentioned culture is not particularly limited to methods that can be produced by expressing an antibody that specifically binds to lipocotin 1 of the present invention or an antigen-binding fragment thereof. Specifically, the above-mentioned culture can be carried out continuously in a batch process or a fed batch or repeated fed batch process, but is not limited thereto.

[0114] Additionally, the step of obtaining an antibody that specifically binds to the lipocotin 1 or an antigen-binding fragment thereof from the culture may be performed by a method known in the art. Specifically, the obtaining method is not particularly limited to methods that can obtain an antibody that specifically binds to the lipocotin 1 of the present invention or an antigen-binding fragment thereof produced therefrom. Preferably, the obtaining method may be a method such as centrifugation, filtration, extraction, spraying, drying, evaporation, precipitation, crystallization, electrophoresis, fractional dissolution (e.g., ammonium sulfate precipitation), chromatography (e.g., ion exchange, affinity, hydrophobicity, and size exclusion).

[0115] Uses of anti-lipocotin 1 antibodies

[0116] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising, as an active ingredient, an antibody that specifically binds to lipocotin 1 or an antigen-binding fragment thereof. The lipocotin 1-specific antibody or the antigen-binding fragment thereof is as described above.

[0117] As used herein, the term "cancer" refers to a disease caused by cells having aggressive characteristics of dividing and proliferating beyond normal growth limits, invasive characteristics of infiltrating surrounding tissues, and metastatic characteristics of spreading to other parts of the body, and is used interchangeably with "malignant tumor."

[0118] Cancer may be selected from the group consisting of gastric cancer, liver cancer, lung cancer, non-small cell lung cancer, colorectal cancer, bladder cancer, bone cancer, blood cancer, breast cancer, melanoma, thyroid cancer, parathyroid cancer, bone marrow cancer, rectal cancer, throat cancer, laryngeal cancer, esophageal cancer, pancreatic cancer, tongue cancer, skin cancer, toenail tumor, uterine cancer, head or neck cancer, gallbladder cancer, oral cancer, pro-anal cancer, colon cancer, and central nervous system tumors, but is not limited thereto.

[0119] Furthermore, "treatment of cancer" refers to inhibiting or preventing the growth of cancer cells or tissues. This concept includes reducing cancer growth and metastasis compared to when the condition is left untreated, as well as reducing resistance to anticancer drugs to enhance therapeutic efficacy. The aforementioned metastasis refers to the process by which tumor (cancer) cells spread to distant parts of the body. "Resistance to anticancer drugs" refers to the absence of therapeutic effect from the beginning of treatment when treating cancer patients with anticancer drugs, or the loss of therapeutic effect over time despite an initial effect. "Prevention" refers to any act of suppressing the occurrence of cancer or delaying its onset through the administration of the aforementioned pharmaceutical composition.

[0120] In the pharmaceutical composition for cancer prevention or treatment of the present invention, the anti-lipocotin 1 antibody or its antigen-binding fragment may be included in any amount (effective amount) depending on the use, formulation, purpose of combination, etc., as long as it can exhibit anticancer activity. Here, "effective amount" refers to the amount of an active ingredient capable of inducing an anticancer effect. Such an effective amount may be determined experimentally within the ordinary capabilities of a person skilled in the art. The pharmaceutical composition of the present invention may contain the antibody or its antigen-binding fragment as an active ingredient in an amount of about 0.1% to about 90% by weight, specifically about 0.5% to about 75% by weight, more specifically about 1% to about 50% by weight, based on the total weight of the composition.

[0121] Pharmacokinetic parameters such as bioavailability and underlying parameters such as clearance rate can also influence efficacy. Therefore, "enhanced efficacy" (e.g., improvement in efficacy) can be attributed to enhanced pharmacokinetic parameters and enhanced efficacy, and can be measured by comparing parameters such as clearance rate and treatment or improvement of cancer in test animals or human subjects.

[0122] The pharmaceutical composition of the present invention may include a conventional and non-toxic pharmaceutically acceptable carrier that is formulated into a preparation according to a conventional method.

[0123] The above-mentioned pharmaceutically acceptable carrier may be any non-toxic substance suitable for delivery to a patient. Distilled water, alcohol, fats, waxes, and inert solids may be included as carriers. Pharmacologically acceptable adjuvants (buffers, dispersants) may also be included in the pharmacological composition.

[0124] As used herein, the term “pharmaceuticalally acceptable carrier” refers to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include saline solution, sterile water, Ringer’s solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, provided that they are sterile and biocompatible. Additionally, other conventional additives such as sweeteners, solubilizing agents, wetting agents, emulsifiers, isotonic agents, absorbents, antioxidants, preservatives, lubricants, fillers, buffers, and bacteriostatic agents may be added as needed.

[0125] The compositions of the present invention may be prepared in various formulations for parenteral administration (e.g., intramuscular, intravenous, or subcutaneous injection). When the pharmaceutical compositions of the present invention are prepared in parenteral formulations, they may be formulated in the form of injectables, transdermal agents, nasal inhalants, and suppositories in accordance with methods known in the art, together with a suitable carrier. Injectable formulations include sterile aqueous solutions, water-insoluble agents, suspensions, emulsions, lyophilized formulations, and suppositories. As water-insoluble agents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrogol, Tween 61, cacao oil, laurin oil, glycerogelatin, etc., may be used. Meanwhile, injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0126] Regarding the formulation of pharmaceutical compositions, it is known in the art, and specifically, reference may be made to literature [Remington's Pharmaceutical Sciences (19th ed., 1995)], etc. The said literature is considered to be part of this specification.

[0127] The antibody or composition of the present invention may be administered to a patient in a therapeutically effective or pharmaceutically effective amount.

[0128] As used herein, the term "administration" means introducing a specific substance into an individual by an appropriate method, and the route of administration of the composition may be any general route as long as it can reach the target tissue. It may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, topically, intranasally, or rectally, but is not limited thereto.

[0129] Here, "therapeutically effective amount" or "pharmaceuticalally effective amount" refers to an amount of a composition effective for preventing or treating a target disease, meaning an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment and that does not cause side effects. The level of the said effective amount may be determined based on factors including the patient's health status, type and severity of the disease, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and elimination rate, duration of treatment, drugs used in combination or concurrently, and other factors well known in the medical field. Specifically, the said therapeutically effective amount refers to an amount of drug effective for treating cancer.

[0130] Specifically, the dosage of the composition of the present invention may vary depending on the patient's age, gender, and body weight. Generally, about 0.1 mg to about 1,000 mg or about 5 mg to about 200 mg per kg of body weight may be administered daily or every other day, or divided into 1 to 3 doses per day. However, the dosage may be increased or decreased depending on the route of administration, severity of the disease, gender, body weight, age, etc., so the scope of the present invention is not limited thereto.

[0131] The subjects to whom the above pharmaceutical composition can be applied (prescribed) are mammals and humans, and it is particularly preferable for humans.

[0132] The antibody of the present invention or a pharmaceutical composition containing the same may be administered as an individual therapeutic agent or in combination with other therapeutic agents, administered sequentially or simultaneously with conventional therapeutic agents, and administered as a single or multiple doses. In this case, the other therapeutic agent may additionally include any compound or natural extract known to have anticancer activity and whose safety has already been verified, in order to enhance or reinforce anticancer activity. It is important to administer an amount that obtains maximum effect with the minimum amount and with minimal or no side effects, taking into account all of the above factors, and this can be easily determined by a person skilled in the art.

[0133] Another aspect of the present invention provides the use of an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, or a pharmaceutical composition comprising the same, for manufacturing a drug for the prevention or treatment of cancer.

[0134] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1, or a pharmaceutical composition containing the same, for preventing or treating cancer.

[0135] Another aspect of the present invention provides a method for preventing or treating cancer comprising the step of administering to an individual an antibody that specifically binds to lipocotin 1, an antigen-binding fragment thereof, or a pharmaceutical composition containing the same.

[0136] At this time, the lipocotin 1-specific antibody or its antigen-binding fragment, pharmaceutical composition, administration, cancer, prevention and treatment are as described above.

[0137] The above individual may be a mammal, preferably a human. Additionally, the above individual may be a patient with cancer or an individual with a high likelihood of developing cancer.

[0138] The route of administration, dosage, and frequency of administration of the above antibody may be administered to the subject in various ways and amounts depending on the patient's condition and the presence or absence of side effects, and the optimal method of administration, dosage, and frequency of administration can be selected within an appropriate range by a person skilled in the art. The preferred dosage of the above anti-lipocotin 1 antibody or its antigen-binding fragment may be administered daily or every other day, or divided into 1 to 3 doses per day, in an amount of about 0.1 mg to about 1,000 mg or about 5 mg to about 200 mg per kg of body weight, depending on the patient's condition, body weight, sex, age, severity of the patient, and route of administration. Such dosages shall not be construed as limiting the scope of the present invention in any aspect.

[0139] In addition, the anti-lipocotin 1 antibody or its antigen-binding fragment may be administered in combination with any compound or natural extract known to have an anticancer therapeutic effect, or formulated in the form of a combination preparation with other drugs.

[0140] Antibody-drug conjugate that specifically binds to lipocotin 1

[0141] Another aspect of the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1; and an anticancer agent bound thereto.

[0142] At this time, the antibody or its antigen-binding fragment that specifically binds to lipocotin 1 is as described above.

[0143] As used herein, the term “antibody-drug conjugate (ADC)” refers to a therapeutic agent in which an antibody and a drug are chemically bound to exhibit a high anticancer effect. In this case, the antibody and the drug may be covalently bound through a linker.

[0144] At this time, the linker may be bound to an antibody that specifically binds to lipocotin 1 or to an antigen-binding fragment thereof. Specifically, a drug may be bound to the antibody by binding a thiol group present in the light or heavy chain of the antibody to a reactive group present in the linker. At this time, an example of a reactive group of the linker may be a maleimide group. Additionally, since the thiol group is contained in cysteine, the linker may be bound to a site on the antibody where cysteine ​​is present.

[0145] In one embodiment, the antibody-drug conjugate may have the structure of the following structural formula I:

[0146] <Structural Formula I>

[0147] Ab-[LD]n

[0148] At this time, in the above structural formula I,

[0149] Ab is an antibody or its antigen-binding fragment that specifically binds to lipocotin 1, and

[0150] L is a linker or direct link,

[0151] D is an anticancer drug, and

[0152] n is an integer from 1 to 10, and specifically, n can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0153] At this time, the antibody or its antigen-binding fragment that specifically binds to the above-mentioned lipocotin 1 is as described above.

[0154] Linker

[0155] As used herein, the term "linker" refers to a component of an antibody-drug conjugate that links an antibody or its antigen-binding fragment to a drug (or payload) through chemical bonding. The linker can covalently bind the antibody and the drug.

[0156] At this time, the linker may be a cleavable linker or a non-cleavable linker.

[0157] As used herein, the term "non-cleaving linker" refers to a linker in which a drug is released through catabolism by cytoplasmic or lysosomal hydrolases after an antibody-drug conjugate is internalized into a target cell.

[0158] In one embodiment, the non-cleaving linker may be a maleimide linker. The maleimide linker may be, for example, a maleimidocaproyl (MC) linker or a succimidyl 4-(N-aleimidomethyl)cyclohexane-1-carboxylate (SMCC) linker.

[0159] In this specification, the term "cleavage linker" refers to an antibody-drug conjugate in which a drug is released by cleavage of the linker. The cleavage linker may be classified according to the principle by which cleavage occurs.

[0160] The above linker may be chemically cleavable or enzymatically cleavable.

[0161] In one embodiment, the chemical cleavage type linker may be an acid-labile linker or a reducible linker.

[0162] The term "acid-labile linker" as used in this specification refers to a linker that is stable at a neutral pH, such as blood, but undergoes cleavage by acid hydrolysis in an acidic environment (pH 5 to 6), such as the cancer cell microenvironment, lysosomes, or endosomes. The acid-labile linker includes a hydrazone linker or an ester linker.

[0163] As used herein, the term "reducible linker" refers to a linker that is reduced by a reducing agent and undergoes cleavage. In one embodiment, the reducible linker may be a disulfide linker.

[0164] The term “enzyme-cleaving linker” as used herein means a linker that is cleaved by a reducing agent within a cell. The enzyme-cleaving linker may be a peptide-based linker or a specific enzyme-based linker.

[0165] As used herein, the term "peptide-based linker" refers to a linker that is cleaved by an enzyme that is relatively abundant within a cell and comprises a specific peptide bond cleavage site. The peptide-based linker may be a linker comprising one or more binding sites selected from the group consisting of valine-citrulline, valine-alanine, and phenylalanine-glycine. In one embodiment, it may be one or more selected from the group consisting of a valine-citrulline linker, a valine-alanine linker, and phenylalanine-glycine.

[0166] As used herein, the term “specific enzyme-based linker” means a linker designed to be cleaved by a specific enzyme. The specific enzyme-based linker may be cleaved by one or more enzymes selected from the group consisting of β-galactoside, β-glucuronide, and phosphodiester. In one embodiment, the specific enzyme-based linker may be one or more selected from the group consisting of β-galactoside linkers, β-glucuronide linkers, and phosphodiester linkers.

[0167] In one embodiment, the cleavage linker may be any one selected from the group consisting of a hydrazone linker, an ester linker, a disulfide linker, a valine-citruline linker, a valine-alanine linker, a phenylalanine-glycine linker, a β-galactoside linker, a β-glucuronide linker, and a phosphodiester linker.

[0168] In one embodiment, the linker may include a structural modification. In one embodiment, the linker may have polyethylene glycol (PGE) substituted in some of its structure or may include a self-immolative chemical structure (SIG).

[0169] Preferably, the antibody-drug conjugate of the present invention may include a non-cleaving linker. Specifically, it may include a maleimide linker.

[0170] Preferably, the antibody-drug conjugate of the present invention may include an enzyme-cleaving linker. Specifically, it may be a β-galactoside linker.

[0171] anticancer drugs

[0172] As used herein, the term "anticancer agent" collectively refers to all drugs effective in treating cancer. In the present invention, the anticancer agent may include any anticancer agent capable of serving as the payload of an antibody-drug conjugate. In one embodiment, the anticancer agent may be a cytotoxic agent.

[0173] As used in this specification, the term "cytotoxic agent" is also referred to as an antitumor drug or a chemotherapy agent. The cytotoxic agent is a general term for drugs that exhibit anticancer activity by acting directly on DNA to block DNA replication, transcription, and translation processes, or by interfering with the synthesis of nucleic acid precursors in metabolic pathways and inhibiting cell division. In this case, the cytotoxic agent may be selected from the group consisting of maytansinoid, auristatin, and calicheamicin cytotoxic agents.

[0174] In one embodiment, the cytotoxic agent may be selected from the group consisting of Ansamitocin, Mertansine, Ravtansine, Monomethyl auristatin E (MMAE), Monomethyl auristatin F (MMAF), and Calicheamicin γ1.

[0175] The above Ansamitocin may be named Ansamitocin P-3 or Maytansinol isobutyrate. In this case, the above Ansamitocin may comprise the structure of the following chemical formula I or a pharmaceutically acceptable salt thereof:

[0176] <Chemical Formula I>

[0177] .

[0178] The above Mertansine may be named Maytansinoid DM1 or N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine. In this case, the Mertansine may comprise the structure of the following chemical formula II or a pharmaceutically acceptable salt thereof:

[0179] Chemical Formula II

[0180] .

[0181] The above Ravtansine is N2'-deavetyl-N2'-(4-mercapto-4-methyl-1-oxopentyl)-maytansine, and may comprise the structure of Formula III below or a pharmaceutically acceptable salt thereof:

[0182] Chemical Formula III

[0183] .

[0184] The above Monomethyl auristatin E(MMAE) refers to (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamido)butanamide. In this case, the above Monomethyl auristatin E(MMAE) may comprise the structure of Chemical Formula IV below or a pharmaceutically acceptable salt thereof:

[0185] <Chemical Formula IV>

[0186] .

[0187] The above Monomethyl auristatin F(MMAF) refers to (S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamido)butanamido)-3-methoxy-5-methylheptanoyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-3-phenylpropanoic acid. In this case, the above Monomethyl auristatin F(MMAF) may comprise the structure of the following chemical formula V or a pharmaceutically acceptable salt thereof:

[0188] Chemical Formula V

[0189] .

[0190] The above Calicheamicin γ1 may comprise the structure of the following chemical formula VI or a pharmaceutically acceptable salt thereof:

[0191] Chemical Formula VI

[0192] .

[0193] Specific example of an antibody-drug conjugate that specifically binds to lipocotin 1

[0194] In the present invention, the antibody-drug conjugate may be an antibody or its antigen-binding fragment that specifically binds to lipocotin 1 according to the present invention, and an anticancer agent MMAF connected by a specific enzyme-based linker, a β-galactoside linker. Specifically, the antibody-drug conjugate may be represented by the following chemical formula VII.

[0195] <Chemical Formula VII>

[0196]

[0197] In the above chemical formula VII, X is NR 1 , O, S or Se, and R 1 H or C 1-8Hydrocarbyl, mAb is an antibody or antigen-binding fragment thereof that specifically binds to the lipocotin 1 according to the present invention, and n is an integer from 1 to 10.

[0198] In one embodiment of the present invention, the antibody-drug conjugate is formed by connecting an antibody that specifically binds to lipocotin 1 and MMAF via a maleimide linker, and it was confirmed that it can specifically target cancer cells overexpressing lipocotin 1 and has a high anticancer effect due to the action of the antibody and the anticancer agent (Figs. 13a and 13b).

[0199] Uses of antibody-drug conjugates

[0200] Another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising the antibody-drug conjugate. The antibody-drug conjugate, cancer, prevention, and treatment are the same as those described above.

[0201] In the pharmaceutical composition for cancer prevention or treatment of the present invention, the antibody-drug conjugate may be included in any amount (effective amount) depending on the use, formulation, purpose of combination, etc., as long as it can exhibit anticancer activity. Here, "effective amount" refers to the amount of an active ingredient capable of inducing an anticancer effect. Such an effective amount may be determined experimentally within the ordinary capabilities of a person skilled in the art. The pharmaceutical composition of the present invention may contain an antibody-drug conjugate as an active ingredient in an amount of about 0.1% to about 90% by weight, specifically about 0.5% to about 75% by weight, more specifically about 1% to about 50% by weight, based on the total weight of the composition.

[0202] Pharmacokinetic parameters such as bioavailability and underlying parameters such as clearance rate can also influence efficacy. Therefore, "enhanced efficacy" (e.g., improvement in efficacy) can be attributed to enhanced pharmacokinetic parameters and enhanced efficacy, and can be measured by comparing parameters such as clearance rate and treatment or improvement of cancer in test animals or human subjects.

[0203] The pharmaceutical composition of the present invention may include a conventional and non-toxic pharmaceutically acceptable carrier that is formulated into a preparation according to a conventional method.

[0204] The above-mentioned pharmaceutically acceptable carrier may be any non-toxic substance suitable for delivery to a patient. A pharmaceutically acceptable adjuvant (buffer, dispersant) may also be included in the pharmacological composition.

[0205] As used herein, the term “pharmaceuticalally acceptable carrier” refers to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include saline solution, sterile water, Ringer’s solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, provided that they are sterile and biocompatible. Additionally, other conventional additives such as solubilizing agents, isotonic agents, antioxidants, preservatives, and buffers may be added as needed.

[0206] The compositions of the present invention may be prepared in various formulations for parenteral administration (e.g., intramuscular, intravenous, or subcutaneous injection). When the pharmaceutical compositions of the present invention are prepared in parenteral formulations, they may be formulated in the form of injectables, transdermal agents, nasal inhalants, and suppositories in accordance with methods known in the art, together with a suitable carrier. Injectable formulations include sterile aqueous solutions, water-insoluble agents, suspensions, emulsions, lyophilized formulations, and suppositories. As water-insoluble agents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrogol, Tween 61, cacao oil, laurin oil, glycerogelatin, etc., may be used. Meanwhile, injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0207] Regarding the formulation of pharmaceutical compositions, it is known in the art, and specifically, reference may be made to literature [Remington's Pharmaceutical Sciences (19th ed., 1995)], etc. The said literature is considered to be part of this specification.

[0208] The antibody-drug conjugate or composition of the present invention may be administered to a patient in a therapeutically effective or pharmaceutically effective amount.

[0209] As used herein, the term "administration" means introducing a specific substance into an individual by an appropriate method, and the route of administration of the composition may be any general route as long as it can reach the target tissue. It may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, topically, intranasally, or rectally, but is not limited thereto.

[0210] Here, "therapeutically effective amount" or "pharmaceuticalally effective amount" refers to an amount of a composition effective for preventing or treating a target disease, meaning an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment and that does not cause side effects. The level of the said effective amount may be determined based on factors including the patient's health status, type and severity of the disease, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and elimination rate, duration of treatment, drugs used in combination or concurrently, and other factors well known in the medical field. Specifically, the said therapeutically effective amount refers to an amount of drug effective for treating cancer.

[0211] Specifically, the dosage of the composition of the present invention may vary depending on the patient's age, gender, and body weight, and generally, about 0.01 mg to about 1,000 mg or about 0.5 mg to about 200 mg per kg of body weight may be administered daily or every other day, or divided into 1 to 3 doses per day. However, since the dosage may be increased or decreased depending on the route of administration, severity of the disease, gender, body weight, age, etc., the scope of the present invention is not limited thereto.

[0212] The subjects to whom the above pharmaceutical composition can be applied (prescribed) are mammals and humans, and it is particularly preferable for humans.

[0213] The antibody of the present invention or a pharmaceutical composition containing the same may be administered as an individual therapeutic agent or in combination with other therapeutic agents, administered sequentially or simultaneously with conventional therapeutic agents, and administered as a single or multiple doses. In this case, the other therapeutic agent may additionally include any compound or natural extract known to have anticancer activity and whose safety has already been verified, in order to enhance or reinforce anticancer activity. It is important to administer an amount that obtains maximum effect with the minimum amount and with minimal or no side effects, taking into account all of the above factors, and this can be easily determined by a person skilled in the art.

[0214] Another aspect of the present invention provides the use of the antibody-drug conjugate or a pharmaceutical composition containing the same for preventing or treating cancer.

[0215] Another aspect of the present invention provides a method for treating or preventing cancer comprising the step of administering the antibody-drug conjugate or a pharmaceutical composition containing the same to an individual. The antibody-drug conjugate, pharmaceutical composition, cancer, prevention, treatment, and administration are the same as described above. In particular, the cancer may be one in which lipocotin-1 is overexpressed.

[0216] The above individual may be a mammal, preferably a human. Additionally, the above individual may be a patient with cancer or an individual with a high likelihood of developing cancer.

[0217] In addition, the antibody-drug conjugate or the pharmaceutical composition containing it may be administered in combination with any compound or natural extract known to have an anticancer therapeutic effect, or may be combined with other drugs.

[0218] Epitope of anti-lipocotinic 1 antibody

[0219] Another aspect of the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to the region from the 27th amino acid to the 185th amino acid of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO. 206.

[0220] Specifically, the antibody or its antigen-binding fragment may specifically bind to a region from the 136th to the 162nd amino acid of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO. 206. More specifically, the antibody or its antigen-binding fragment may specifically bind to any one amino acid residue selected from the group consisting of the 136th (T136), 141st (L141), 149th (I149), 150th (R150), 157th (R157), 159th (E159), 160th (L160), 162nd (R162) amino acids of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO. 206 and combinations thereof. More specifically, the antibody or its antigen-binding fragment may specifically bind to any one amino acid residue selected from the group consisting of the 136th (T136), 141st (L141), 157th (R157), 159th (E159), 160th (L160), 162nd (R162) amino acids of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO. 206, and combinations thereof. Preferably, the antibody or its antigen-binding fragment may specifically bind to the 157th (R157) amino acid residue of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO. 206.

[0221] The above antibody or its antigen-binding fragment may compete with one or more of the following antibodies or their antigen-binding fragments for binding to the lipocotin 1 protein.

[0222] The term "competes for binding" in the above competitive binding assay refers to the ability of an antibody or another antigen-binding moiety to interfere with the binding of another antibody or antigen-binding moiety to a specific antigen. The ability or degree to which an antibody or another antigen-binding moiety can interfere with the binding of another antibody or antigen-binding moiety to a specific antigen, and accordingly, whether it may be referred to as mutually competing according to the present invention, can be determined using a standard competitive binding assay. One suitable assay may involve the use of ViaCore technology, which can measure the degree of interaction using surface plasmon resonance technology. Other assays for measuring mutual competition may use ELISA-based methods. In the present invention, the term "competitive binding" may mean that the antibody or its antigen-binding fragment competes for the Lipocotin 1 protein by interfering with the binding of the following antibody or its antigen-binding fragment to the Lipocotin 1 protein. Such competitive binding may mean sharing a binding site (epitope) for the lipocotin 1 protein.

[0223] The antibody or its antigen-binding fragment that competitively binds to the above antibody or its antigen-binding fragment for lipocotin 1 protein comprises: a heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 222, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228; an antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 222, HCDR2 represented by the amino acid sequence of SEQ ID NO. 223, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 226, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 229;An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 224, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 230; or may include a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228.

[0224] The present invention will be described in more detail below through examples. These examples are solely for illustrating the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be interpreted as being limited by these examples.

[0225] Example 1. Preparation and production of Lipocotin 1 antigen

[0226] To select antibodies that bind to lipocortin 1 (lipocortin 1; LPC-1; annexin A1; ANXA1; AA1, P04083), human antigen proteins were produced on GenScript via GomaBio translation (#U0348GD200).

[0227] A gene fragment encoding human lipocotin 1 antigen protein (LPC-1-his, SEQ ID NO. 207), which does not code for the 2nd to 26th amino acids at the N-terminus and contains tags for asparagine from the 27th to 376th serine and six histidine at the C-terminus, was loaded into a pET-30a(+) vector (GenScript) using NdeI (#R0131, NEB) and HindIII (#R3104, NEB) restriction enzymes.

[0228] The above vector was transformed into Escherichia coli BL21-Gold (DL3) (#C606010, Thermo) to express the lipocotin 1 protein, and then purified using a His-tag affinity column (#17-5247-01, Cytiva) and ion exchange chromatography (SP Sepharose column). Mouse lipocotin 1 antigen (mLPC-1-his, P10107) and monkey lipocotin 1 antigen (moLPC-1-his, F7D2H1) were synthesized and produced by TriOar Inc. using the same method as described above. The antigen proteins expressed and purified by the above method are shown in Figures 1a and 1b.

[0229] In addition, a mutant lipocotin 1 antigen was constructed to be used for the analysis of the antigen-binding sites (epitopes) of antibodies that specifically bind to the human lipocotin 1 antigen. After obtaining gene fragments encoding mutant lipocotin 1 through gene synthesis or PCR, they were cloned into pcDNA vectors using SfiI (#FD1824, Thermo) and NheI (#FD0974, Thermo) restriction enzymes. The mutation information of the antigen is listed in Table 1.

[0230] LPC-1 Antigen Mutation Information Antigen Name | Antigen Type | Gene Domain | Mutation Site WT (SEQ No. 239) LPC-1-hisAA1 (NP000691) S27-N346 M1~K26 deletion (Common to all antigens below) D1 (SEQ No. 240) LPC-1-D1-C-hisAA1+AA2 fusion | S27-N346 Substitution of AA1's S27-K113 with AA2's S18-K104 D2 (SEQ No. 241) LPC-1-D2-C-hisAA1+AA2 fusion | S27-N347 Substitution of AA1's T114-K185 with AA2's T105-K176 D4 (SEQ No. 242) LPC-1-D3-C-hisAA1+AA2 fusion | S27-N348 AA1's Substitution of G186-C271 with AA2's G177-C262 D4 (Sequence No. 243) LPC-1-D4-C-hisAA1+AA2 fusion S27-D347 Substitution of AA1's A272-N347 with AA2's I263-D338 V1 (Sequence No. 244) LPC-1-V1-C-hisAA1 S27-N346 N43D, S45E, S46 RV2 (Sequence No. 245) LPC-1-V2-C-hisAA1 S27-N346 V48A, A49L, A50N, L51I, H52E, K53 TV3 (Sequence No. 246) LPC-1-V3-C-hisAA1 S27-N346 M56K, V57 TV4 (Sequence No 247)LPC-1-V4-C-hisAA1S27-N346A63V, I66V, D67N, K71NV5(Sequence No. 248)LPC-1-V5-C-hisAA1S27-N346Q79D, K81A, A82FV6(Sequence No. 249)LPC-1-V6-C-hisAA1S27-N346L85Q, Q86R, E87R, G89K, P91EV7(Sequence No. 250)LPC-1-V7-C-hisAA1S27-N346D93A, E94S, K98SV8(Sequence No. 251)LPC-1-V8-C-hisAA1S27-N346E106T, V108I, A110GV9(Sequence No 252)LPC-1-V9-C-hisAA1S27-N346F118Y, D121S, R124K,A126SV10(Sequence No. 253)LPC-1-V10-C-hisAA1S27-N346T136S, L141IV11(Sequence No. 254)LPC-1-V11-C-hisAA1S27-N346I149L, R150QV12(Sequence No. 255)LPC-1-V12-C-hisAA1S27-N346R157K, E159M, L160Y, R162TV13(Sequence No. 256)LPC-1-V13-C-hisAA1S27-N346A165E, T169IV14(Sequence No. 257)LPC-1-V14-C-hisAA1S27-N346A179L, L180M, L181VV10_1(Sequence No. 258)LPC-1-V10_1-C-hisAA1S27-N346T136SV10_2(Sequence No. 259)LPC-1-V10_2-C-hisAA1S27-N346L141IV12_1(Sequence No. 260)LPC-1-V12_1-C-hisAA1S27-N346R157KV12_2(Sequence No. 261)LPC-1-V12_2-C-hisAA1S27-N346E159MV12_3(Sequence No. 262)LPC-1-V12_3-C-hisAA1S27-N346L160YV12_4(Sequence No 263)LPC-1-V12_4-C-hisAA1S27-N346R162T,

[0231] At this time, the above AA2 may be ANXA2 and may include the amino acid sequence of SEQ ID NO. 264.

[0232] The antigen mutant protein expressed and purified by the above method is shown in Fig. 1c.

[0233] Example 2. Biopanning of a scFv-type antibody phage library binding to Lipocotin 1

[0234] To screen antibody fragments that bind to lipocotin 1, biopanning was performed in two ways (immunoassay tubes and beads) using the fully human antibody phage library (Ymax®-ABL) of Y-Biologics (PA21298, PA21299).

[0235] Human lipocotinoid 1 antigen protein (LPC-1-his) was coated onto immunosorbent tubes or His purification beads and blocked (4% skim milk / 0.01% Tween-20 / PBS). 7.5×10 10 Human antibody phage libraries in the form of scFv (single chain variable fragment) with diversity and M1 helper phages (Y-Biologics) were prepared by infecting Escherichia coli (XL1-Blue, Y-Biologics) and incubating at 37°C for 16 hours, followed by centrifugation to dissolve the supernatant in PBS buffer (#LB001-02, Welgene). The prepared phage libraries were added to antigen-coated immunosorbent tubes or His purification beads and reacted at room temperature for 2 hours. After washing (0.01% Tween-20 / PBS), positive phages specifically bound to the antigen were extracted. These positive phages were reinfected into Escherichia coli, and panning was performed three times to increase the number of washes and amplify the phages, thereby obtaining poly-positive phages.

[0236] Escherichia coli infected with poly-positive phages were cultured on solid media (#2× YT-GMC, Y-Biologics), and a total of 1,000 colonies were selected from the formed colonies and cultured in 96-deep-well plates with 2× YT-GMC medium and M1 helper phage. Next, the cultured product was centrifuged to extract the supernatant containing mono-scFv phages, and an enzyme-linked immunosorbent assay (ELISA) was performed.

[0237] Specifically, the supernatant containing mono-positive phages was placed into a tube coated with LPC-1-his antigen (human lipocotinic 1 antigen protein). At this time, the supernatant of a phage control without target specificity (#38 or AB#38, Y-Biologics) was also added separately for testing. ELISA was performed under conditions coated with non-specific antigen controls (#ITGA6-Fc, #CD58-Fc, #RAGE-Fc, #AITR-Fc, Y-Biologics) to select a total of 452 individual clones that specifically bind to LPC-1.

[0238] The gene sequences of 452 selected individual clones were analyzed to identify 100 hit clones with different sequences. In addition, Direct ELISA was performed to exclude clones with a binding value (OD 450 nm) of less than 3 for LPC-1-his or a binding value of 0.3 or higher for non-specific antigens, ultimately securing 97 hit clones (Figs. 2a to 3b).

[0239] VL shuffling library and VH CDR mutation library for PA21299-010F10 hit clone were prepared through a commissioned service (ES2301000014) by Y-BioLogics Co., Ltd., and biopanning (beads) was performed twice (CPA23009, CPA23010) in the same manner as in Example 2 above to finally select four antibody clones (H2I23DGA, H2I23DID, H2I23EOA, and H2I23EOB) with mature affinities.

[0240] Meanwhile, to improve the physical properties and enhance the antigen-specific affinity of TRO22H2I using the Triore library, biopanning was performed several times in the same manner as in Example 2 above. 258 phage-hit clones were selected by modifying the amino acid sequence of the structural region (FR) within the heavy chain variable region (VH) or the amino acid sequence of the light chain variable region (VL) of TRO22H2I. Among these, two clones (TAB0035, TAB0041) with significantly improved productivity due to modification of the structural region (FR) were secured. Biopanning was performed on these two clones by modifying the heavy chain complementarity determining region (CDR) and changing the light chain variable region.

[0241] As a result, four antibody clones (H2I24095, H2I24109, H2I24128, H2I24132) with improved physical properties and affinity were selected from 13 novel sequences in which excellent antigen binding ability was confirmed.

[0242] The sequences of each of the above antibody clones are listed in Tables 2 to 5.

[0243] Example 3. Sequence analysis of antibody fragments binding to lipocotin 1

[0244] Among the 97 antibody fragment hit clones obtained through the screening of Example 2 above, the amino acid sequences of 30 antibody clones and 4 antibody clones with mature affinity were analyzed. Table 2 shows the amino acid sequence of the heavy chain complementarity determining region (HCDR) of the heavy chain variable region (VH), and Table 3 shows the amino acid sequence of the light chain complementarity determining region (LCDR) of the light chain variable region (VL). Tables 4 and 5 show the amino acid sequences of the heavy chain variable region (VH) and light chain variable region (VL), respectively.

[0245] Heavy chain complementation determining region of scFv binding to lipocotin 1 protein IgG CODEHCDR1HCDR2HCDR3TRO22O0DGFTFDDYA(SEQ No. 1)INWDSSNI(SEQ No. 23)VKLASDSGGYFES(SEQ No. 42)TRO22O3AGFTFNMHP(SEQ No. 2)IKYDGSK(SEQ No. 24)SCNGLGQDDEAFDI(SEQ No. 43)TRO22O4AGFTFDDYG(SEQ No. 3)ISWNSGSI(SEQ No. 25)TRYLHRLPFDV(SEQ No. 44)TRO22O6EGFTFSDFA(SEQ No. 4)ISGTSGRT(SEQ No. 26)ARGSPSGSADY(SEQ No. 45)TRO22O9AGITFGDYA(SEQ No. 5)IRSKAYGGTT(SEQ No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22O9BGYTLTELS(Sequence No. 6)FDLRNGET(Sequence No. 28)ARSYYDILTGYYRDAFDI(Sequence No. 47)TRO22O9GGNTFNGYY(Sequence No. 7)INPSGSST(Sequence No. 29)AQEGWTEQHPYAEYLQN(Sequence No. 48)TRO22O9HGFTFGDYA(Sequence No. 8)IRSKSFGGTT(Sequence No. 30)ARDGNYYDSRDYYYDTFDL(Sequence No. 49)TRO22A0GGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22A0HGFSFSDHY(Sequence No. 9)IRSAAYGGTT(Sequence No. 31)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22A2CGFTFSDYY(Sequence No. 10)ISSSGSTI(Sequence No. 32)ARATYYDILTGYYGYFDY(Sequence No. 50)TRO22A2DGFNFNTYA(Sequence No. 11)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22A2FGFTFGDYA(Sequence No. 8)IRTKTYGGTP(Sequence No. 33)VRDGNYYDSRGYYYDTFDL(Sequence No. 51)TRO22A2GGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No27)ARDGNYYDSRGYYYDTFDM(Sequence No. 52)TRO22A3BGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22A3CGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G1AGFTFSDHY(Sequence No. 12)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G1BGFSFGDYA(Sequence No. 13)IRSKRFGGTT(Sequence No. 34)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G1EGFNFGDHA(Sequence No. 14)IRSKTYGGTP(Sequence No. 35)ARDGNYYDSRGYYYDTFDM(Sequence No. 52)TRO22G1FGFIFGDYA(Sequence No. 15)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G2BGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G2CGFTFSDHY(Sequence No. 12)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G2GGFTFGDYA(Sequence No. 8)IRSKAYGGTT(Sequence No. 27)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G2HGFTFGEYA(Sequence No. 16)IRSKAFGGTP(Sequence No. 36)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G3EGFSFSSHA(Sequence No. 17)ISETGGIA(Sequence No. 37)ARDGNYYDSRGYYYDTFDM(Sequence No. 52)TRO22G4AGFTTNSYT(Sequence No. 18)ISSSTNYM(Sequence No. 38)ARDGNYYDSRGYYYDTFDM(Sequence No. 52)TRO22G4HGFTFGDYY(Sequence No. 19)ISSSSAYT(Sequence No. 39)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G5HGFTLDDYS(Sequence No. 20)IYTGGNT(Sequence No40)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)TRO22G8AGYTFSSYW(Sequence No. 21)INPGNGHT(Sequence No. 41)ARDGNYYDSRGYYYDTFDM(Sequence No. 52)TRO22H2IGFNFGDYP(Sequence No. 22)IRSKSFGGTT(Sequence No. 30)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I23DGAGFNFGDYP(Sequence No. 22)IRSKSFGGTT(Sequence No. 30)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I23DIDGFNFGDYP(Sequence No. 22)IRDKRIGGTT(Sequence No. 214)ARDGNYYDSRGYYYDTFDL(Sequence No 46)H2I23EOAGFNFGDYP(Sequence No. 22)IRDKTVGGTT(Sequence No. 215)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I23EOBGFNFGDYP(Sequence No. 22)IRPKWLGGTT(Sequence No. 216)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I24095GFNFGDYP(Sequence No. 22)IRDKQTGGTT(Sequence No. 222)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I24109GFNFGDYP(Sequence No. 22)IRDRLIGGTT(Sequence No. 223)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I24128GFNFGDYP(Sequence No 22)IRDKLTGGTT(Sequence No. 224)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)H2I24132GFNFGDYP(Sequence No. 22)IRDKRIGGTT(Sequence No. 214)ARDGNYYDSRGYYYDTFDL(Sequence No. 46)

[0246] Light chain complementation determining region of scFv binding to lipocotin 1 protein IgG CODEL CDR1LCDR2LCDR3TRO22O0DSGSIASKY(SEQ No. 53)EDNQR(SEQ No. 73)QSYDSSSVV(SEQ No. 85)TRO22O3ASGSIASKY(SEQ No. 53)EDNQR(SEQ No. 73)QSYDSSSVV(SEQ No. 85)TRO22O4ASGSIASKY(SEQ No. 53)EDNKR(SEQ No. 74)QSYDDNTVV(SEQ No. 86)TRO22O6ESGSIASKY(SEQ No. 53)EDNQR(SEQ No. 73)QSYDSSTVV(SEQ No. 87)TRO22O9AQAISSW(SEQ No. 54)ATS(SEQ No. 75)EQAHSFPLT(Sequence No. 88)TRO22O9BRSVLHSSSNQNY(Sequence No. 55)WAS(Sequence No. 76)QQCYSTPYT(Sequence No. 89)TRO22O9GQYIGTY(Sequence No. 56)DAS(Sequence No. 77)QQSYITPFA(Sequence No. 90)TRO22O9HQDISRW(Sequence No. 57)AAS(Sequence No. 78)EQAHSFPLT(Sequence No. 88)TRO22A0GQSLLHSNGYNY(Sequence No. 58)LDS(Sequence No. 79)MQTLHTLYT(Sequence No. 91)TRO22A0HQGISSY(Sequence No. 59)VAS(Sequence No. 80)QQSYRTPYT(Sequence No. 92)TRO22A2CQSISNW(Sequence No 60)GAS(Sequence No. 81)QQSYITQTWT(Sequence No. 93)TRO22A2DQSLLHSNGYNY(Sequence No. 58)LGS(Sequence No. 82)MQALQTPRT(Sequence No. 94)TRO22A2FQGISNW(Sequence No. 61)AAS(Sequence No. 78)QQGHSFPYT(Sequence No. 95)TRO22A2GQNLLHSNGYNY(Sequence No. 62)MGS(Sequence No. 83)MQPLQIPFT(Sequence No. 96)TRO22A3BQDIGHW(Sequence No. 63)AAS(Sequence No. 78)QQANSFPYT(Sequence No. 97)TRO22A3CQNLLHSNGYNY(Sequence No. 62)LGS(Sequence No. 82)MQTLQTLYT(Sequence No 98)TRO22G1AQSISNY(Sequence No. 64)AAS(Sequence No. 78)QQSYNTPRT(Sequence No.99)TRO22G1BQPISTW(Sequence No. 65)AAS(Sequence No. 78)QQANSFPYT(Sequence No. 97)TRO22G1EQSLLHSNGYNY(Sequence No. 58)LGS(Sequence No. 82)MQTLQTLYT(Sequence No. 98)TRO22G1FQDIKNY(Sequence No. 66)GAS(Sequence No. 81)QQYEELAALT(Sequence No. 100)TRO22G2BQGISTW(Sequence No. 67)AAS(Sequence No. 78)QQSYSIPRT(Sequence No. 101)TRO22G2CQGIRNY(Sequence No. 68)AAS(Sequence No. 78)LQDHDYPYT(Sequence No. 102)TRO22G2GQSLLHSNGYNY(Sequence No 58) LGS (Sequence No. 82) MQSLQTPPYT (Sequence No. 103) TRO22G2HHDISSW (Sequence No. 69) ATS (Sequence No. 75) QQGKSFPYT (Sequence No. 104) TRO22G3EQSLLHTNGYNY (Sequence No. 70) SAS (Sequence No. 84) MQALQTPWT (Sequence No. 105) TRO22G4AQSLLHSNGYNY (Sequence No. 58) LGS (Sequence No. 82) MQTLQTLPYT (Sequence No. 106) TRO22G4HQSLLHSNGYYY (Sequence No. 71) LGS (Sequence No. 82) MQGLMTPFT (Sequence No. 107) TRO22G5HQSLLHSNGYNY (Sequence No. 58) LGS (Sequence No 82)MQALRTPYT(Sequence No. 108)TRO22G8AQSLLHSNGYNY(Sequence No. 58)LGS(Sequence No. 82)MQALRTPYT(Sequence No. 108)TRO22H2IQGIRSW(Sequence No. 72)AAS(Sequence No. 78)QQANSFPYT(Sequence No. 97)H2I23DGAQSLLHSNGYNY(Sequence No. 58)MGS(Sequence No. 83)MQALQTQLT(Sequence No. 217)H2I23DIDQGIRSW(Sequence No. 72)AAS(Sequence No. 78)QQANSFPYT(Sequence No. 97)H2I23EOAQGIRSW(Sequence No. 72)AAS(Sequence No. 78)QQANSFPYT(Sequence No 97)H2I23EOBQGIRSW(Sequence No. 72)AAS(Sequence No. 78)QQANSFPYT(Sequence No. 97)H2I24095QGVRTW(Sequence No. 225)ASS(Sequence No.227)QQAYSFPFT(Sequence No. 228)H2I24109QGISRW(Sequence No. 226)AAS(Sequence No. 78)QQSYSTPRT(Sequence No. 229)H2I24128QGIRSW(Sequence No. 72)AAS(Sequence No. 78)QQANSFPRT(Sequence No. 230)H2I24132QGVRTW(Sequence No. 225)ASS(Sequence No. 227)QQAYSFPFT(Sequence No. 228)

[0247] The heavy chain variable region of scFv binding to lipocotin 1 protein IgG CODEVHTRO22O0DQMQLVQSGGGLVQPGRSLRLSCAVSGFTFDDYAMHWVRQAPGKGLEWVSGINWDSSNIGYADSVKGRFTISRDNAKNSLYLQMNNLRSEDTAVYYCVKLASDSGGYFESWGQGTLVTVSS(SEQ No. 109)TRO22O3AQVQLVQSGGGVVQPGRSLKLSCETSGFTFNMHPMHWVRQAPGKGLEWVASIKYDGSKRYYTDSVKGRFTVSRDHVDTVYLQMNSLRAEDTAVYYCSCNGLGQDDEAFDIWGQGTMVTVSS(SEQ No. 110)TRO22O4AQVQLVESGGGLVQPGRSLRLSCAASGFTFDDYGMHWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNSLRSEDTAVYYCTRYLHRLPFDVWGKGTTVTVSS (SEQ ID NO: 111)TRO22O6EQMQLVESGGGLVQPGGSLRLSCAASGFTFSDFAMTWVRQAPGKGLEWVSSISGTSGRTYYADSVKGRSTVSTDNSKNTVYLRMNSLRAEDTAVYYCARGSPSGSADYWGQGTLVTVSS (SEQ ID NO: 112)TRO22O9AQVQLVQSGGGLVQPGRSLRLSCTASGITFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS (SEQ ID NO: 113)TRO22O9BQMQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDLRNGETIYAQRFQGRVTMSGDTATDTAYMELSSLRSEDTAVYYCARSYYDILTGYYRDAFDIWGQGTMVTVSS (SEQ ID NO:114)TRO22O9GQMQLVQSGAEVQKPGASVKVSCKASGNTFNGYYIHWVRQAPGQGLEWMGIINPSGSSTFYAQKFRGRVTMTKDTSTSTVYMKLSSLRSEDTAVYYCAQEGWTEQHPYAEYLQNWGQGTLVTVSS(서열번호 115)TRO22O9HQVQLVESGGGLVQPGRSLRLSCTASGFTFGDYAMSWVRQAPGKGLEWIGFIRSKSFGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRDYYYDTFDLWGQGTMVTVSS(서열번호 116)TRO22A0GQVQLVQSGGGLVKPGRSLRLSCTASGFTFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTTVTVSS(서열번호 117)TRO22A0HQVQLVESGGGLVPPGGSLRLSCAASGFSFSDHYMDWVRQAPGKGLEWVGFIRSAAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 118)TRO22A2CQVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSYISSSGSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARATYYDILTGYY'-GYFDYWGQGTLVTVSS(서열번호 119)TRO22A2DQMQLVESGGGLVQPGRSLRLSCAASGFNFNTYAMHWVRQAPGRGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호120)TRO22A2FQVQLVESGGGLIQPGRSLRLSCTTSGFTFGDYAMSWVRQAPGKGLEWVGFIRTKTYGGTPEYAASLKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCVRDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 121)TRO22A2GQVQLVESGGGLIQPGRSLRLSCTASGFTFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDMWGQGTLVTVSS(서열번호 122)TRO22A3BQVQLVQSGGGLVQPGRSLRLSCTASGFTFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 123)TRO22A3CQVQLVESGGGLVKPGRSLRLSCTASGFTFGDYAMSWFRQAPGKGLEWVGFIRSKAYGGTTEHAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 124)TRO22G1AQVQLVQSGGGLVQPGGSLRLSCAASGFTFSDHYMDWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 125)TRO22G1BQVQLVESGGGLIQPGRSLRLSCRTSGFSFGDYAMTWVRQAPGKGLEWVGFIRSKRFGGTTEYAASVKGRFTISRDDSTSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호126)TRO22G1EQVQLVESGGGLVQPGHSLRLSCTSSGFNFGDHAVSWVRQAPGKGLEWVGFIRSKTYGGTPEYAASVKDRFSISRDDSKSSAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDMWGQGTLVTVSS(서열번호 127)TRO22G1FQVQLVESGGGLVQPGRSLRLSCTASGFIFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTDYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 128)TRO22G2BQMQLVQSGGGLVKPGRSLRLSCTASGFTFGDYAMSWFRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 129)TRO22G2CQMQLVESGGGLVQPGGSLRLSCAASGFTFSDHYMDWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 130)TRO22G2GQVQLVESGGGLVQPGRSLRLSCTASGFTFGDYAMSWVRQAPGKGLEWVGFIRSKAYGGTTEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 131)TRO22G2HQMQLVESGGDLVQPGRSLRLSCRTSGFTFGEYAMSWVRQAPGKGLEWVGFIRSKAFGGTPEYAASVKGRFTISRDDSKSIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호132)TRO22G3EQVQLVESGGGLIHPGESLRLSCAASGFSFSSHAMSWVRQAPGKGLEWVATISETGGIAFYADSVKGLFTISRDNSKNTLSLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDMWGQGTVVTVSS(서열번호 133)TRO22G4AQVQLVESGGGWVQPGGSLRLSCAGSGFTTNSYTMAWVRQAPGKGLEWVSSISSSTNYMYYADSVKGRFTISRDNAKNSVYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDMWGQGTLVTVSS(서열번호 134)TRO22G4HQVQLVESGGGLVKPGGSLRLSCAASGFTFGDYYMNWIRQAPGKGLEWVAYISSSSAYTRYADSVKGRFTISRDNTKNSLYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 135)TRO22G5HQVQLVESGGVAVQPGGSLRLSCTASGFTLDDYSMHWVRQRPGKGLEWVSVIYTGGNTYYSESVKGRFNIARDNSKNSLYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMVTVSS(서열번호 136)TRO22G8AQVQLVQSGAEVKKPGASVKLSCKASGYTFSSYWMHWVRQAPGQRLEWMGEINPGNGHTNYNEKFKSRVTITVDKSASTAYMELSSLRSEDTAVYYCARDGNYYDSRGYYYDTFDMWGQGTLVTVSS(서열번호 137)TRO22H2IQVQLVQSGGGLVQPGRSLRISCKASGFNFGDYPLSWVRQAPGKGLEWIGFIRSKSFGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호138)H2I23DGAQVQLVQSGGGLVQPGRSLRISCKASGFNFGDYPLSWVRQAPGKGLEWIGFIRSKSFGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 138)H2I23DIDQVQLVQSGGGLVQPGRSLRISCKASGFNFGDYPLSWVRQAPGKGLEWIGFIRDKRIGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 218)H2I23EOAQVQLVQSGGGLVQPGRSLRISCKASGFNFGDYPLSWVRQAPGKGLEWIGFIRDKTVGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 219)H2I23EOBQVQLVQSGGGLVQPGRSLRISCKASGFNFGDYPLSWVRQAPGKGLEWIGFIRPKWLGGTTEYAASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTMITVSS(서열번호 220)H2I24095QVQLVQSGGGLVQPGGSLRLSCAASGFNFGDYPLSWVRQAPGKGLEWIGFIRDKQTGGTTEYADSVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTLVTVSS(서열번호 231)H2I24109QVQLVQSGGGLVQPGGSLRLSCAASGFNFGDYPLSWVRQAPGKGLEWIGFIRDRLIGGTTEYADSVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTLVTVSS(서열번호232)H2I24128QVQLVQSGGGLVQPGGSLRLSCTASGFNFGDYPMSWVRQAPGKGLEWIGFIRDKLTGGTTEYVASVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTLVTVSS(서열번호 233)H2I24132QVQLVQSGGGLVQPGGSLRLSCTASGFNFGDYPMSWVRQAPGKGLEWIGFIRDKRIGGTTEYATSVKGRFTISRDDSRDIAYLQMNSLRAEDTAVYYCARDGNYYDSRGYYYDTFDLWGQGTLVTVSS(서열번호 234)

[0248] The variable region of the light chain of scFv binding to lipocotin 1 protein IgG CODEVLTRO22O0DQLVLTQPHSVSESPGKTVTISCARSSGSIASKYVQWYQQRPGSAPTAVIYEDNQRPSGVPDRFSGSIDTSSNSASLTISGLKTEDEADYYCQSYDSSSVVFGGGTKLTVL(SEQ No. 139)TRO22O3AQLVLTQPHSVSESPGKTVTISCTRSSGSIASKYVQWYQQRPGSAPTTVIYEDNQRPSGVPDRFSGSIDISSNSASLTISGLKTEDEADYYCQSYDSSSVVFGGGTKLTVL(SEQ No. 140)TRO22O4ANFMLTQPHSVSESPGKMVTISCTRSSGSIASKYVQWYQQRPGSAPITVIYEDNKRPSGVPDRFSGSIDSFSNSASLTITGLKTEDEADYYCQSYDDNTVVFGGGTKLTVL (SEQ ID NO: 141)TRO22O6EQLVLTQPHSVSESPGKTVTISCTRSSGSIASKYVQWYQQRPGSAPTTVIYEDNQRPSGVPDRFSGSIDISSNSASLTISGLKTEDEADYYCQSYDSSTVVFGGGTKLTVL (SEQ ID NO: 142)TRO22O9ADIQMTQSPSSVFASVGDRVTITCRASQAISSWLAWYQQKPGKAPKLLIYATSYLQTGVPSRFSGSGSGTDFTLSINNLQPEDSATYFCEQAHSFPLTFGPGTKVDIK (SEQ ID NO: 143)TRO22O9BDIQMTQSPDSLAVSLGERATINCNSSRSVLHSSSNQNYLAWYQQKPGQPPKLLISWASGRQSGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQCYSTPYTFGQGTKLDIK (SEQ ID NO: 144)TRO22O9GDIQMTQSPSFLSASVGDRVTITCRASQYIGTYLAWYQQQPGKAPKLLIYDASTLQPGVPSRFSGGGSGTDFTLTIRSVQPEDYATYFCQQSYITPFAFGQGTKLEIK (SEQ ID NO:145)TRO22O9HDIQMTQSPSSVSASVGDRVTITCRASQDISRWLAWYQQKPGKAPKLLIYAASILQTGVPSRFSGSGSGTDFTLSINNLQPEDSATYFCEQAHSFPLTFGPGTKVDIK(서열번호 146)TRO22A0GDIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLDSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLHTLYTFGQGTKLDIK(서열번호 147)TRO22A0HDIQMTQSPSSLSASVGDKVTITCRASQGISSYLAWYQQKPGKAPKLLIYVASSLQTGVPSRFSGSRSGTDFTLTISSLQSEDFGTYYCQQSYRTPYTFGQGTKLEIK(서열번호 148)TRO22A2CDIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYGASNLKSGVPSRFSGSGSGTHFTLTISSLHPEDFATYYCQQSYITQTWTFGQGTKVEIK(서열번호 149)TRO22A2DDIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPRTFGQGTKVDIK(서열번호 150)TRO22A2FDIQMTQSPSSVSASVGDRVTITCRASQGISNWVAWYQQKPGKAPKLLIYAASTLHSGVPSRFSGSASGTDFTLTISSLQPEDFAIYYCQQGHSFPYTFGQGTKLEIK(서열번호 151)TRO22A2GDIVMTQTPLSLPVTPGEPAFISCRSSQNLLHSNGYNYLDWYLQKPGQSPQLLIYMGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQPLQIPFTFGGGTKVEIK(서열번호152)TRO22A3BDIQMTQSPSSVSASLGDRVTITCRASQDIGHWLAWYQQKPGKAPKHLIYAASSLHSGVPSRFSGSGSGTEFTLTISNLQPEDFATYYCQQANSFPYTFGQGTKVDIK(서열번호 153)TRO22A3CDIVMTQTPLSLPVTPGEPASISCRSSQNLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLQTLYTFGQGTKVEIK(서열번호 154)TRO22G1ADIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLMSAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPRTFGQGTKVDIK(서열번호 155)TRO22G1BDIQMTQSPSSVSASVGDRVTITCRASQPISTWLVWYQHKPEKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTINSLQPDDFATYYCQQANSFPYTFGQGTKLDIK(서열번호 156)TRO22G1EDIVMTQTPLSLPVIPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLQTLYTFGQGTKLEIK(서열번호 157)TRO22G1FDIQMTQSPSSLSASVGDRITISCQASQDIKNYLNWYQKKPGKAPKLLIYGASNLAAGVPSRFSGSGYGTDFTFIISGLQPEDVATYYCQQYEELAALTFGGGTKVEIK(서열번호 158)TRO22G2BDIQMTQSPSSVSASIGDRITITCRASQGISTWLAWYQQKPGKAPKLLIYAASSLHSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSYSIPRTFGQGTKVEIK(서열번호159)TRO22G2CDIQMTQSPSSLSASVGDRVTITCRASQGIRNYLGWYQQKPGRAPKLLIYAASSLQSGVPSRFRGSGSGTDFTLTISSLQPEDFATYYCLQDHDYPYTFGQGTKVEIK(서열번호 160)TRO22G2GDIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLEWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQSLQTPPYTFGQGTKVEIK(서열번호 161)TRO22G2HDIQMTQSPSSVSASVGDRVTITCRASHDISSWLAWYQQKPGKAPKLLIYATSSLQNGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQGKSFPYTFGQGTKVEIK(서열번호 162)TRO22G3EDIVMTQTPLSLPVTLGEPASISCRSSQSLLHTNGYNYLDWYLQKPGQSPQLLVYSASTRASGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCMQALQTPWTFGQGTKVEIK(서열번호 163)TRO22G4ADIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVQAEDVGTYYCMQTLQTLPYTFGQGTKLEIK(서열번호 164)TRO22G4HDIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYYYLDWYLQKPGQSPQVLIYLGSNRAPGVPDRFSGSGSGTDFTLKISRVEADDVGIYYCMQGLMTPFTFGPGTKLDIK(서열번호 165)TRO22G5HDIVMTQTPLSLPVTPGGPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALRTPYTFGQGTKVDIK(서열번호166)TRO22G8ADIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLGWYRQKPGQSPQLLIYLGSNRAFGVPDRFSGSGSGTDYTLKISRVEAEDVGVYYCMQALRTPYTFGQGAKLDIK(서열번호 167)TRO22H2IDIQMTQSPSSVSASVGDRVTLTCRASQGIRSWLAWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFILTINSLQPDDFATYYCQQANSFPYTFGQGTKVEIK(서열번호 168)H2I23DGADIVMTQTPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYMGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTQLTFGGGTKVEIK(서열번호 221)H2I23DIDDIQMTQSPSSVSASVGDRVTLTCRASQGIRSWLAWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFILTINSLQPDDFATYYCQQANSFPYTFGQGTKVEIK(서열번호 168)H2I23EOADIQMTQSPSSVSASVGDRVTLTCRASQGIRSWLAWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFILTINSLQPDDFATYYCQQANSFPYTFGQGTKVEIK(서열번호 168)H2I23EOBDIQMTQSPSSVSASVGDRVTLTCRASQGIRSWLAWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFILTINSLQPDDFATYYCQQANSFPYTFGQGTKVEIK(서열번호 168)H2I24095DLVMTQSPSSVSASVGDKVTISCRASQGVRTWLAWYQQQPGKAPKLLIYASSKLQSGVPSRFSGSGSGTDFALTISSVQPEDFGTYYCQQAYSFPFTFGQGTRLEIK(서열번호235)H2I24109DIVMTQSPSSVSASVGDRVTITCRASQGISRWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPRTFGQGTKLEIK (SEQ ID NO: 236)H2I24128DIQMTQSPSSVSASVGDRVTITCRASQGIRSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPRTFGQGTKVEIK (SEQ ID NO: 237) H2I24132DIVMTQSPSSVSASVGDKVTISCRASQGVRTWLAWYQQQPGKAPKLLIYASSKLQSGVPSRFSGSGSGTDFALTISSVQPEDFGTYYCQQAYSFPFTFGQGTRVEIK (SEQ ID NO: 238)

[0249] Example 4. Production of IgG in the form of a human antibody binding to lipocotin 1

[0250] A recombinant vector was constructed to convert 97 scFv-type heat clones into human antibody-type IgG. PCR primers (Table 6) were produced by Bionia Inc. and used for PCR (#639298, Takara).

[0251] Specifically, the heavy chain variable region was inserted into the pC3.1-HC vector (TriOar) using restriction enzymes SfiI (#R0123S, NEB) and NheI-HF (#R3131, NEB), and the light chain variable region was transferred into the pC3.1-LC vector (TriOar) using EagI-HF (#R3505, NEB) and BglII (#R0144, NEB) to construct each recombinant vector. Each recombinant gene sequence was verified using a sequencing service (Bioneer).

[0252] PRIMER 5' → 3' direction[VH: variable heavy chain, VL: variable light chain, F: forward, R: reverse]VHF01tctttttggtggccacagcggccgatgtccactcgcaggtgcagctggtggag(Sequence No. 169)VHF02tctttttggtggccacagcggccgatgtccactcgcaggtgcagctggtacag(Sequence No. 170)VHF03tctttttggtggccacagcggccgatgtccactcgcaggtgcagctggtagag(Sequence No. 171)VHF04tctttttggtggccacagcggccgatgtccactcgcagatgcagctggtgcag(Sequence No. 172)VHF05tctttttggtggccacagcggccgatgtccactcgcaggtgcagctgcaggag(Sequence No. 173)VHF06tcttttggtggccacagcggccgatgtccactcgcagatgcagctggtagag(Sequence No. 174)VHF07tcttttggtggccacagcggccgatgtccactcgcaggtgcagctggtgcag(Sequence No. 175)VHF08tctttttggtggccacagcggccgatgtccactcgcagatgcagctggtggag(Sequence No. 176)VHF09tcttttggtggccacagcggccgatgtccactcgcagatgcagctggtacag(Sequence No. 177)VHF10tctttttggtggccacagcggccgatgtccactcgcatgtgcagctggtggag(Sequence No. 178)VHF11tctttttggtggccacagcggccgatgtccactcgcaggtacagctgcagcag(Sequence No. 179)VHRatgggcccttggtgctagctgaggagacggtga(Sequence No. 180)VLF01tctttttggtggccacagcggccgatgtccactcgtcctatgagctgactcag(Sequence No.181) VLF02tctttttggtggccacagcggccgatgtccactcgaattttatgctgactcag (SEQ ID NO: 182) VLF03tctttttggtggccacagcggccgatgtccactcgcagctcgtgctgactcag (SEQ ID NO: 183) VLF04tctttttggtggccacagcggccgatgtccactcgtcctatgagctgacacag (SEQ ID NO: 184) VLF05tctttttggtggccacagcggccgatgtccactcggatattgtgatgacccag (SEQ ID NO: 185) VLF06tctttttggtggccacagcggccgatgtccactcggacatccagatgacccag (SEQ ID NO: 186)VLF07tctttttggtggccacagcggccgatgtccactcgcagtctgccctgactcag(Sequence No. 187)VLF08tctttttggtggccacagcggccgatgtccactcgaattttatgctgactcaa(Sequence No. 188)VLF09tctttttggtggccacagcggccgatgtccactcgtcctatgagttgacacag(Sequence No. 189)VLF10tctttttggtggccacagcggccgatgtccactcgcaggctgtggtgactcag(Sequence No. 190)VLR01atggtgcagccacagatcttaggacggtcagcttggt(Sequence No. 191)VLR02atggtgcagccacagatcttaggacggtgaccttggt(Sequence No. 192)VLR03atggtgcagccacagatcttttgatttccactttggt(Sequence No. 193)VLR04atggtgcagccacagatcttttgatttccagcttggt(Sequence No. 194)VLR05atggtgcagccacagatcttttgatatccaccttggt(Sequence No. 195)VLR06atggtgcagccacagatcttttgatctccactttggt(Sequence No. 196)VLR07atggtgcagccacagatcttttgatctccaccttggt(Sequence No.197)VLR08atggtgcagccacagatcttttgatatccactttggt(Sequence No. 198)VLR09atggtgcagccacagatcttttgatctccagcttggt(Sequence No. 199)VLR10atggtgcagccacagatcttttgatttccaccttggt(Sequence No. 200)VLR11atggtgcagccacagatcttttgatatccagcttggt(Sequence No. 201)VLR12atggtgcagccacagatcttttgatttctagcttggt(Sequence No. 202)VLR13atggtgcagccacagatcttttgatatctagcttggt(Sequence No. 203)VLR14atggtgcagccacagatcttttgatttccagtcgtgt(Sequence No. 204)VLR15atggtgcagccacagatcttttgatatccagcttggc(Sequence No. 205)

[0253] Example 5. Production of IgG in the form of a human antibody binding to lipocotin 1

[0254] 97 types of IgG antibodies were produced through research services by Cytiva Korea.

[0255] The recombined pair of pC3.1-HC and pC3.1-LC vectors are ExpiCHO TM The vectors were expressed and purified in ExpiCHO-S cell lines using the Expression system kit (#A29133, Thermo). For example, equal amounts of the two vectors were mixed and transformed according to the manufacturer's instructions. The cells were then cultured in a 25 mL scale (110 rpm, 5% CO2, 32°C) while measuring cell count (Luna II, Logos) and antibody levels (Cedex Bio, Roche). When the defined criteria were met (viability >70% or antibody level >150 mg / L), the cells were centrifuged (5,000 rpm, 4°C, 30 min), and only the supernatant was harvested after processing through a 0.22 μm filter. The cells were then processed on an AKTA PCC75 (Cytiva) instrument using a Protein A affinity column (HiTrap TMFibro PrismA, Cytiva) and desalination column (HiTrap) TM Ninety-four types of antibodies, excluding three types (TRO22G2A, TRO22O0C, TRO22O1D), were purified using Desalt XK16 (Cytiva) (1 mg to 5 mg). The purity of the antibodies was confirmed by SEC-HPLC (KW803 column, Waters Alliance) analysis (Figs. 4a to 4d).

[0256] In addition, eight selected clones with mature affinity were prepared in IgG form and purified using the same method as in Examples 4 and 5. The purified products were analyzed by SDS-PAGE (Figs. 4e and 4f).

[0257] Example 6. Selection of Lipocotin 1-specific binding antibodies via ELISA evaluation

[0258] The binding ability of the 94 types of IgG antibodies produced by the method of Example 5 above to antigens was confirmed through ELISA (enzyme-linked immunosorbent assay).

[0259] Each antigen was prepared by diluting it in PBS solution (#P0750-N10L, Biowest) to a concentration of 0.1 to 1.0 μg / mL. As human antigens, LPC-1-his (a form lacking 26 amino acids from the N-terminus (#U0348GD200, GenScript), AA-1 (full-length annexin A1) (#NBP2-52001, Novus), AA-2 (full-length annexin A2) (#ab93005, Abcam), and AA-5 (full-length annexin A5) (#AN5-H5123, Acrobio) were used. mLPC-1-his (TriOar) and moLPC-1-his (TriOar) were used as mouse and monkey antigens, respectively.

[0260] Each IgG antibody was prepared by diluting with a diluent (4% skim milk / 0.05% Tween-20 / PBS).

[0261] 0.1 mL of the prepared antigen solution was dispensed into each well of a 96-well plate (#15142, Thermo) and reacted using a Multi-shaker (#3D-300, FinePCR) at room temperature for 1 hour. Then, the plates were washed twice with 0.2 mL of wash solution (0.05% Tween-20 / PBS). Next, 0.1 mL of each diluted hit antibody was added to each well and reacted at room temperature for 1 hour, followed by two washes. Human IgG (#31154, Thermo) was used as a negative control. The test antibody (#31413, Thermo) was diluted to a ratio of 1:3000, dispensed 0.1 mL into each well, and reacted by shaking at room temperature for 1 hour. Afterward, the plates were washed twice, and 0.1 mL of TMB substrate solution (#T0440, Sigma) was added and reacted in the dark. When color development occurred, 0.05 ml of 1 N sulfuric acid solution (#S1408, Samchun) was added to stop the reaction, and the absorbance was measured at 450 nm using a spectrometer (#GM3000, Promega) (Figs. 5a, 5b and Table 7).

[0262] As a result, as shown in Figures 5a and 5b, IgG antibodies were found to have various binding abilities to human lipocotin 1 antigens (N-terminal 26-amino acid deletion antigen (LPC-1-his), full-length antigen (AA1)). In addition, as shown in Table 7, antibodies with cross-binding ability to mouse lipocotin 1 (mLPC-1-his) and antibodies that selectively bind or have cross-binding ability to family members (AA-2, AA-5) were obtained.

[0263] The binding affinity of the above-mentioned HEAT antibody at different concentrations was analyzed using the Graphpad Prism 9.4 program with the above measurements. As a result, the binding affinity of the above-mentioned HEAT antibody to human antigen (LPC-1-his) and mouse antigen (mLPC-1-his) increased in a concentration-dependent manner (Figs. 6a and 6b).

[0264] In addition, for the four antibodies with mature affinities, the binding affinity increased in a concentration-dependent manner for each antigen (LPC-1-his, mLPC-1-his, moLPC-1-his) at a low concentration (0.1 μg / mL) (Fig. 6c). It was confirmed that the four antibodies selectively bind to the full-length antigen (AA-1) and family members (AA-2, AA-5) (Fig. 6d). Furthermore, the concentration-dependent binding affinity of the four antibodies with improved productivity and mature affinities to the human antigen (LPC-1-his) was confirmed (Fig. 6e). Through these results, it was confirmed that the hit antibody and the antibodies with mature affinities specifically bind to the lipocotin 1 antigen and can cross-bind to mouse antigens or monkey antigens.

[0265] Specific and selective binding affinity of IgG forms (ELISA) at OD 450 nm; [-]: <0.1, [+]: 0.1–0.5, [++]: 0.5–1.0, [+++]: >1.0, N / D: Not determined IgG CODE mLPC-1-his,[0.1] µg / mL AA-2,[0.1] µg / mL AA-5,[0.1] ㎍ / ㎖TRO22A0G[+++][-][-]TRO22A0H[+++][-][-]TRO22A1B[+++][-][-]TRO22A2C[+++][-][-]TRO22A2D[ +++][-][-]TRO22A2F[+++][-][-]TRO22A2G[+++][-][-]TRO22A3B[++][-][-]TRO22A3C[+++][-][-]TRO2 2F8CN / D[-][-]TRO22G0D[+++]N / DN / DTRO22G0E[+++][-][-]TRO22G1A[++][-][-]TRO22G1B[+++][-][-] TRO22G1E[+++][-][-]TRO22G1F[+++][-][-]TRO22G2B[++][-][-]TRO22G2C[+++][-][-]TRO22G2G[+++][ -][-]TRO22G2H[+++][-][-]TRO22G3E[+++][-][-]TRO22G4A[+++][-][-]TRO22G4H[+++][-][-]TRO22G5 H[+++][-][-]TRO22G8A[+++][-][-]TRO22H2I[+++][-][-]TRO22H4C[+++][-][-]TRO22O0D[+++][-][-]T RO22O3A[+++][+++][+++]TRO22O4A[+++][+++][-]TRO22O6E[+++][+++][++]TRO22O6O[+]N / DN / DTRO22O 9A[+++][-][-]TRO22O9B[+++][-][-]TRO22O9G[+++][-][-]TRO22O9H[+++][-][-]TRO22O9O[+++]N / DN / D

[0266] Example 7. Selection of Lipocotin 1-specific binding antibodies through FC evaluation

[0267] The binding affinity of IgG hit antibodies and antibodies with mature affinity to lipocotin 1 expressed in each cell line was confirmed using a flow cytometer (FC; flow cytometry) (CytoFlex S, Beckman).

[0268] HUVEC cell line (human umbilical vein endothelial cell line) was cultured in EGM2 growth medium [EGM(#CC-3162, Lonza) / 10% FBS(#SH30919.03, Hyclone) / 1× antibiotic-antifungal agent(#SV30079.01, Hyclone)], A549 cell line (human lung carcinoma cell line) was cultured in RPMI growth medium [RPMI(#SH30027.01, Hyclone) / 10% FBS / 1× antibiotic-antifungal agent], and NIH-3T3 cell line (mouse fibroblast cell line) was cultured in DMEM growth medium [DMEM(#SH30243.01, Hyclone) / 10% FBS / 1× antibiotic-antifungal agent] (#Steri-Cycle i160CO2 incubator, Forma, 37℃).

[0269] To confirm lipocotin 1 expression in each cell line, antibodies under the following three conditions were used to measure expression using a flow cytometer (CytoFlex S, Beckman) and analyzed using the Kaluza analysis program (Beckman). i) #MAB3770 (R&D) was used as the primary antibody, #14-4714-82 (Invitrogen) as the mouse IgG isotype control, and #17-4015-82 (Invitrogen) as the measurement antibody (Allophycocyanin fluorescence). ii) #71-3400 (Invitrogen) was used as the primary antibody, #31235 (Invitrogen) as the rabbit IgG isotype control, and #A-11008 (Invitrogen) as the measurement antibody (Alexa Fluor 488 fluorescence). iii) The F4 clone sequence of Public Patent 10-2015-0029457 was prepared using the same method as described in Examples 4 and 5 above, and the produced #MA11SS (Cytiva Research Service, TriOar) was used as the primary antibody, #31154 (Invitrogen) as the isotype control for human IgG, and #FI-3000 (Vector) as the measurement antibody (Fluorescein fluorescence).

[0270] Specifically, the cell line cultured by the above method is 5×10⁶ in 500 µl of DPBS (#SH30028.02, Hyclone). 5Cells were suspended and dispensed into sterile reaction tubes (#72.690, Sarstedt) or 96-deep-well plates (#90063, Bioneer), and 500 µl of cold 4% paraformaldehyde (#PC2031-050-00, Biosesang) solution was added and reacted at room temperature for 10 minutes. Afterward, the supernatant was removed by centrifugation (Avanti J-15R, Beckman, 4℃, 1,000 rpm, 3 min), and the supernatant was removed by washing twice with 500 µl of washing solution (2% FBS / DPBS) to prepare the cell pellet.

[0271] Flow cytometry analysis under fixation conditions was performed using the following method.

[0272] The primary antibody, diluted 1:50 in 100 µl of washing solution, and each isotype control or IgG-type hit antibody were added to the prepared cell pellet and suspended, then reacted at 4°C for 1 hour. After centrifugation, the cells were washed twice and the supernatant was removed. The secondary antibody, diluted 1:50 in 100 µl of washing solution, was added to the cells and suspended, then reacted at 4°C for 1 hour while blocking light. The reaction mixture was then centrifuged, washed twice, and the supernatant removed. The cells were again suspended in 300 µl of DPBS and analyzed using a flow cytometer under light-blocking conditions while maintaining 4°C. The binding affinity at different antibody concentrations was analyzed using Graphpad Prism 9.4 software.

[0273] Flow cytometry analysis under fixation and permeabilization conditions was performed using the following method.

[0274] 1 ml of SAP (0.1% saponin / DPBS) (#A18820.22, Alfa Aesar) solution was added to the prepared cell pellet and suspended, after which the supernatant was removed by centrifugation. The primary antibody diluted (1:50) in 100 µl of SAP solution, each isotype control, or IgG-type hit antibody were added to the cells and mixed, then reacted at 4°C for 1 hour. After centrifugation, the cells were washed twice with 1 ml of SAPF (0.1% saponin / 2% FBS / DPBS) solution, and the supernatant was removed. The secondary antibody diluted (1:50) in 100 µl of SAP solution was added to the cells and suspended, then reacted at 4°C for 1 hour while blocking light. Then, the cells were centrifuged and washed once each with 1 ml of SAPF solution and 1 ml of washing solution, after which the supernatant was removed. The cells were then suspended again in 300 µl of DPBS and analyzed using a flow cytometer under light-shielded conditions at 4°C. The binding affinity at different antibody concentrations was analyzed using the Graphpad Prism 9.4 program.

[0275] As a result, as shown in Figures 7a to 8e, each hit antibody and eight antibodies with matured affinity in cell lines (HUVEC, A549, NIH-3T3) exhibited excellent binding affinity to Lipocotin 1 expressed in each cell. Furthermore, the binding affinity increased in a concentration-dependent manner. Through these results, it was possible to secure antibodies that bind specifically to Lipocotin 1, and it was confirmed that said antibodies can cross-bind to mouse antigens.

[0276] Example 8. Confirmation of binding affinity of lipocotin 1-specific binding antibody

[0277] To measure the binding affinity of IgG-type hit antibodies to lipocotin 1, an OCTET R8 system (Forte) based on BLI (bio-layer interferometry) was used for analysis.

[0278] The Ni-NTA chip (#18-5101, ForteBio) biosensor was stabilized by immersing it in 1×Kinetic buffer (ForteBio) for approximately 10 minutes, after which the LPC-1-his antigen, prepared at a concentration of 2 µg / ml in the buffer, was immobilized onto the biosensor. The antigen not immobilized on the biosensor was washed with the buffer for 5 minutes, followed by an association reaction with IgG antibodies of different concentrations for 10 minutes, and then a dissociation reaction for 10 minutes. For all samples required for analysis, a 96-well black, flat-bottom polypropylene microplate (#655209, GreinerBio) was used. Additionally, all experiments were performed at 30°C and 1,000 rpm, and sensogram data were collected during the binding and dissociation processes over time. The results collected by the above method are applied to a 1:1 global binding fitting model according to OCTET data analysis software to determine the equilibrium dissociation constant (K D ) calculated.

[0279] The analysis process was conducted in three stages: data selection, processing, and analysis. Additionally, among the sample treatment concentrations, those at which the sensitogram was confirmed to be saturated were excluded during the analysis stage. To obtain reliable values ​​(R² > 0.95 and X² < 3), the antibody and antigen concentrations and reaction times were selected under optimized conditions. Binding graphs at different antibody concentrations were analyzed using the Graphpad Prism 9.4 program.

[0280] As a result, as shown in FIGS. 9a to 9h and Table 8, the K of the hit antibodies D The value is 10 -10 It is in the M range, and the K of the four types of antibodies with mature affinities D The value is 10 -11 to <10 -12 It showed excellent binding affinity as a range of M.

[0281] Binding affinity (OCTET) of IgG form hit antibody LPC-1-his onto Ni-NTA chip IgG CODEK D (M)K a (1 / Ms)K dis (1 / s)Full X 2 Full R 2 TRO22O3A3.68×10 -10 1.31 x 10 6 4.82 x 10 -4 0.15950.9968TRO22O4A7.09ⅹ10 -10 6.37 x 10 5 4.51 x 10 -4 0.05700.9974TRO22O6E1.06ⅹ10 -10 7.54 x 10 5 7.95 x 10 -5 0.06770.9973TRO22O9G9.82ⅹ10 -10 1.03 x 10 6 1.01 x 10 -3 0.04240.9939TRO22A0H6.36ⅹ10 -10 6.83 x 10 5 4.34 x 10 -4 0.04930.9961TRO22H2I9.99ⅹ10 -10 1.98 x 10 6 1.98 x 10 -3 0.37790.9886TRO22O9A9.06ⅹ10 -10 1.58 x 10 6 1.43 x 10 -3 0.63890.9881TRO22G4A5.62ⅹ10 -10 1.32 x 10 6 7.44 x 10-4 0.26080.9935H2I23DGA9.13×10 -11 2.22 x 10 5 2.03 x 10 -5 0.81040.9982H2I23DID<1.00ⅹ10 -12 2.87 x 10 5 <1.00 x 10 -7 1.63580.9988H2I23EOA<1.00ⅹ10 -12 3.14 x 10 5 <1.00 x 10 -7 1.32060.9990H2I23EOB7.63×10 -11 3.10 x 10 5 2.37 x 10 -5 1.90910.9987K a (1 / MS): Rate of associationK dis (1 / s): Rate of dissociationK D (M): Affinity constant Full X 2 : The sum of the squared deviationValues ​​close to zero indicate a good curve fit.Full R 2 : R 2 is the coefficient of determination (COD)Values ​​close to 1.0 indicate a good curve fit.

[0282] Meanwhile, to measure the binding affinity of four mature hit antibodies to lipocotin 1, technical services (Z-24-117588) were requested from the K-Bio Institute (KBIO), and binding kinetics analysis was performed using a Biacore T200 instrument (Cytiva) based on surface plasmon resonance (SPR).

[0283] Specifically, each flow cell was activated by flowing a mixture of 100 mM N-hydroxysuccinimide (NHS) and 400 mM 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (mixed in a 1:1 ratio (v / v)) through the Series S CM5 chip for 420 seconds. The capture molecule, diluted with immobilization buffer, was flowed through the flow cell at a flow rate of 10 µl / min for 420 seconds to achieve a volume of 9,000–12,000 RU. Excess carboxyl groups remaining on the surface of the CM5 chip were inactivated by flowing 70 µl of 1 M ethanolamine-HCl (pH 8.0). Each ligand (target protein or LPC-1-his) was diluted in running buffer and injected into the reference and sample flow cells for capture during each cycle. Each analyte was prepared by setting the saturation concentration to the maximum and performing serial dilution six times in twofold increments using running buffer. Ligands and analytes were injected and regenerated using the multi-cycle kinetics method, and sensorograms were measured. Injection time and flow rate were controlled using Biacore T200 control software. The measured sensorograms were fitted to a 1:1 binding model evaluation model using BIAevaluation software (version 3.1). The reliability of the sensorgram fitting was evaluated using Chi2 and U-values ​​according to the manufacturer's manual.

[0284] As a result, as shown in Table 9 and Figures 10a to 10e, the bonding strength (K of TRO22H2I) D) is 10 -9 The level was M, and the four hit antibodies with mature affinity were 10 against the target protein, human LPC-1-his antigen. -10 High binding affinity of M (K D It was confirmed that it represents ).

[0285] Binding affinity (BIACORE) of IgG form hit antibody LPC-1-his onto Series S CM5 chip IgG CODEK D (M)K a (1 / Ms)K dis (1 / s)R max (RU)Chi 2 (RU 2 )U-valueTRO22H2I2.31×10 -9 2.26 x 10 6 5.22 x 10 -3 35.360.831.85H2I240952.04ⅹ10 -10 1.11 x 10 6 2.27 x 10 -4 67.113.628.96H2I241093.37ⅹ10 -10 1.31 x 10 6 4.41 x 10 -4 63.793.314.07H2I241282.22ⅹ10 -10 8.75 x 10 5 1.94 x 10 -4 72.451.716.89H2I241322.09ⅹ10 -10 1.10 x 10 6 2.31 x 10 -4 88.654.446.89K a (1 / MS): Rate of associationK dis (1 / s): Rate of dissociationK D (M): Affinity constant Confidence range: Chi 2 The value is R max 10% or less, U-value 25 or less

[0286] Example 9. Analysis of the antigen binding site of a lipocotin 1-specific binding antibody

[0287] In order to analyze the epitopes, which are the antigen-binding sites of lipocotin 1-specific binding antibodies, mutant antigens of LPC-1 were prepared and enzyme immunoassay (ELISA) was performed in the same manner as in Example 6 above.

[0288] As described in Table 1 above, the amino acid sequences of domains 1 through 2 of the human LPC-1 protein were divided into 14 regions to create group mutations, and the binding of antibodies against the LPC-1 protein was verified by ELISA.

[0289] As a result, it was observed that the antigen binding affinity of the antibody completely disappeared for the V10 mutant antigen in which the 136th threonine and 141st leucine of the antigen protein were substituted, and for the V12 antigen in which the 157th arginine, 159th glutamate, 160th leucine, and 162nd arginine were substituted (Fig. 11b). In addition, through six single amino acid substitution mutations of the V10 and V12 antigens with reduced binding affinity (Table 1), it was confirmed that the 157th arginine is the most critical site for antibody-antigen binding (Fig. 11c).

[0290] Example 10. Analysis of in vitro transport efficiency of lipocotin 1-specific binding antibodies

[0291] To confirm the in vitro transcytosis, or endothelial transcytosis, of IgG hit antibodies under in vitro conditions, human umbilical vein endothelial cell lines (HUVEC) and cancer cell lines were co-cultured.

[0292] 10 µl of gelatin solution (#G1393, Sigma) was added to a Transwell insert (#353095, Falcon) and incubated for 1 hour in a 37°C incubator (#Steri-Cycle i160, Forma). Subsequently, the cells were prepared by irradiating with ultraviolet light for 30 minutes on a sterile workbench (#JSCB-1500SB, JSR). The HUVEC cell line was prepared by adding 1×10⁶ cells to 200 µl of EMG2 growth medium (EGM2 medium / 10% FBS / 1X antibiotic-antifungal agent). 4 The cells were suspended at the cell concentration and inoculated into the insert prepared as described above. The MCF-7 (breast adenocarcinoma) cell line was inoculated at 5×10⁻⁶ per 600 µl of RPMI growth medium (RPMI medium / 10% FBS / 1X antibiotic-antifungal agent). 4 The cells were suspended at a concentration and inoculated into a 24-well bottom plate (#3526, Corning). The transwell inert inoculated with the above HUVEC cell line was placed on top of the bottom-well inoculated with MCF-7, and cultured for 2 days (37°C, 5% CO2). On the 2nd day of culture, the medium of the insert was removed, and 200 µl of EGM2 conditional medium (EGM2 medium / 2% FBS / 1X antibiotic-antifungal agent) was added, followed by an additional 3 days of culture.

[0293] On the 5th day of culture, the medium was removed from the insert, and 200 µl of the prepared sample solution was added for incubation (37°C, 5% CO2). The sample solution was prepared by diluting the IgG isotype control or IgG hit antibody to 67 nM in EGM2 free medium (EGM2 medium / 1× antibiotic-antifungal agent). Between 4 and 24 hours after treatment with the sample solution, a specific amount (10 µl) of medium was collected from the bottom well, and the amount of IgG in the medium was measured using an IgG total Human ELISA kit (#BMS2091, Invitrogen) according to the manufacturer's instructions. Standard quantification curve analysis was performed using the 4PL sigmoidal curve function of the Graphpad Prism 9.4 program.

[0294] As a result, as shown in Figure 12, it was confirmed that the transport efficiency of lipocotin 1-specific binding antibodies was increased 3 to 6 times higher compared to the endothelial cell pass-through rate of the IgG control group.

[0295]

[0296] Example 11. Confirmation of in vivo antitumor efficacy of a lipocotin 1-specific antibody-drug conjugate

[0297] An antibody-drug conjugate (ADC), TRO22H2I-MMAF (test substance), was prepared by conjugating the human anti-lipocotin 1 antibody, TRO22H2I (IgG1, kappa, thiomab), with the maleimide linker-MMAF (Monomethyl auristatin F) using a known method (Nature biotechnology 2008, Vol. 26, 925~932).

[0298] Specifically, 2.9 mg of TRO22H2I antibody (1.53 mg / mL, 1927 µL) was dissolved in 4,000 µL of sodium citrate (#36439.36, Alfaaesar) buffer (25 mM, pH 6.0), and then 2 mM DTT (#43816-10 mL, Sigma) was reacted at 37°C for 3 hours to reduce cysteine ​​residues. The DTT used in the reaction was removed from the reduced antibody using a PD-10 column (#17-0851-01, Cytiva), and the mixture was centrifuged with Vivaspin-6 (#28-9323-17, Cytiva) at 20°C at a speed of 4,000 rpm for 18 minutes. The same process was repeated 6 times to remove any remaining DTT. The recovered antibody at this time was 2.5 mg (380 µl, 6.53 mg / mL), showing a yield of 84%. The recovered antibody was stored at 4°C for 3 days and oxidized again to restore the disulfide bond. The oxidized antibody was treated with maleimide linker-MMAF (Monomethyl auristatin F) at a 6-fold molar concentration and reacted with stirring at 37°C for 3 hours to bind the drug to the cysteine ​​residue of the antibody. Subsequently, to remove the remaining linker-MMAF, PD-10 was used once, followed by Vivaspin-6 (20°C, 4000 rpm, 18 min) seven times. Finally, 2.23 mg (2.3 mg / mL, 970 µl) of antibody-drug conjugate (TRO22H2I-MMAF) was obtained.

[0299] The antitumor efficacy of the MMAF-conjugated test substance in an in vivo JIMT-1 breast cancer tumor model was evaluated through the Asan Medical Center Anticancer Efficacy Evaluation Support Center (APEX) (#ASP-TRO-2213-2).

[0300] Frozen JIMT-1 human breast cancer cell lines held at APEX were thawed and subcultured at 3 to 4-day intervals in a culture medium (DMEM medium / 10% heat-inactivated FBS / 1× antibiotic-antifungal agent) at 37°C and 5% CO2. When cell viability was over 90% after staining with trypan blue, the cells were suspended in cold saline and 5×10⁶ cells were formed. 6 It was prepared for transplantation at a cell / 50 µl / mouse.

[0301] Prepared cells (5×10⁶) were injected into the flank of 6-week-old BALB / c nude mice stabilized for 1 week using a disposable syringe. 6 Cells (50 µl) and 50 µl of Matrigel were mixed and implanted subcutaneously in 100 µl portions. The tumor volume was calculated according to the formula (Volume = (a × 2b) / 2, where a is the short diameter and b is the long diameter), and on the day of administration, mice with a tumor volume of 150 ± 20 mm³ were selected and separated into groups using the paired-matching method.

[0302] The test substance was diluted with a buffer solution (PBS, #SH30256.01, Hyclone) to the administration concentration (0.02 μmol / kg; 3 mg / kg) and administered as a single intravenous (IV) injection. At this time, a buffer solution (PBS, #SH30256.01, Hyclone) was used as a control.

[0303] Tumor volume and clinical symptoms were measured twice a week after drug administration, and TGI values ​​were calculated using the formula (TGI(%) = {1-(△T / △C)}×100). Upon completion of the observation period, the animals were euthanized, and tumor tissues were excised from each individual, weighed, and photographed. All data acquired during the study period were recorded in an Excel format. Additionally, the test substance used was an antibody-drug conjugate (ADC) in which the microtubule inhibitor MMAF was conjugated to a TRO22H2I antibody, which was confirmed to be specific to lipocotin 1 and exhibit excellent binding affinity.

[0304] As a result, as shown in Fig. 13a, tumor growth inhibitory ability was confirmed compared to the negative control group. At this time, there were no abnormal findings in mouse body weight (Fig. 13b).

Claims

1. (a) i) heavy chain complementarity determining region 1 (HCDR1) represented by the amino acid sequence of SEQ ID NO. 210; ii) heavy chain complementarity determining region 2 (HCDR2) represented by the amino acid sequence of SEQ ID NO. 211; and iii) heavy chain variable region (VH) comprising heavy chain complementarity determining region 3 (HCDR3) represented by the amino acid sequence of SEQ ID NO. 212 and (b) iv) a light chain complementarity determining region 1 (LCDR1) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 53 to 72, SEQ ID NOs 225 and 226; v) a light chain complementarity determining region 2 (LCDR2) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 73 to 84 and SEQ ID NOs 227; and vi) an antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region (VL) including a light chain complementarity determining region 3 (LCDR3) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 85 to 108, SEQ ID NOs 217 and SEQ ID NOs 228 to 230; GX1X2X3X4X5X6X7(Sequence No. 210) Here, X1 is F, I, Y or N, X2 is T, S, N or I, X3 is F, L or T, X4 is G, D, N, S or T, X5 is D, M, E, G, T or S, X6 is Y, H, F or L, and X7 is A, P, G, S, Y or W; IRX8X9X 10 X 11 GGTX 12 (Sequence No. 211) Here, X8 is S, T, D, or P, X9 is K, A, or R, and X 10 is A, S, T, R, W, Q or L, and X 11 is Y, F, I, V, L or T, and X 12 is T or P and; X 13 RDGNYYDSRX 14 YYYDTFDX 15 (Sequence No. 212) Here X 13 is A or V, and X 14 is G or D, and X 15 is L or M.

2. In Paragraph 1, The above heavy chain variable region (VH) is i) Heavy chain complementarity determining region 1 (HCDR1) represented by any one amino acid sequence selected from the group consisting of SEQ ID NOs 1 to 22; ii) heavy chain complementarity determining region 2 (HCDR2) represented by any one amino acid sequence selected from the group consisting of SEQ ID NO. 27, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 33 to 36, SEQ ID NO. 214 to 216, and SEQ ID NO. 222 to 224; and iii) an antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a heavy chain complementarity determining region 3 (HCDR3) represented by any one amino acid sequence selected from the group consisting of SEQ ID NO. 46, SEQ ID NO. 49, SEQ ID NO. 51 and SEQ ID NO.

52.

3. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 1, HCDR2 represented by the amino acid sequence of SEQ ID NO. 23, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 42; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

85.

4. In Paragraph 3, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 109; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

139.

5. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 2, HCDR2 represented by the amino acid sequence of SEQ ID NO. 24, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 43; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

85.

6. In Paragraph 5, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 110; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

140.

7. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 3, HCDR2 represented by the amino acid sequence of SEQ ID NO. 25, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 44; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 74, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

86.

8. In Paragraph 7, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 111; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

141.

9. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 4, HCDR2 represented by the amino acid sequence of SEQ ID NO. 26, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 45; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 53, LCDR2 represented by the amino acid sequence of SEQ ID NO. 73, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

87.

10. In Paragraph 9, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 112; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

142.

11. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 5, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 54, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

88.

12. In Paragraph 11, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 113; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

143.

13. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 6, HCDR2 represented by the amino acid sequence of SEQ ID NO. 28, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 47; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 55, LCDR2 represented by the amino acid sequence of SEQ ID NO. 76, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

89.

14. In Paragraph 13, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 114; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

144.

15. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 7, HCDR2 represented by the amino acid sequence of SEQ ID NO. 29, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 48; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 56, LCDR2 represented by the amino acid sequence of SEQ ID NO. 77, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

90.

16. In Paragraph 15, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 115; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

145.

17. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 49; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 57, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

88.

18. In Paragraph 17, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 116; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

146.

19. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 79, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

91.

20. In Paragraph 19, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 117; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

147.

21. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 9, HCDR2 represented by the amino acid sequence of SEQ ID NO. 31, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 59, LCDR2 represented by the amino acid sequence of SEQ ID NO. 80, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

92.

22. In Paragraph 21, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 118; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

148.

23. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 10, HCDR2 represented by the amino acid sequence of SEQ ID NO. 32, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 50; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 60, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

93.

24. In Paragraph 23, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 119; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

149.

25. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 11, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

94.

26. In Paragraph 25, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 120; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

150.

27. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 33, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 51; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 61, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

95.

28. In Paragraph 27, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 121; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

151.

29. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

96.

30. In Paragraph 29, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 122; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

152.

31. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 63, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

32. In Paragraph 31, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 123; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

153.

33. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 62, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

98.

34. In Paragraph 33, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 124; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

154.

35. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 64, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

99.

36. In Paragraph 35, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 125; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

155.

37. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 13, HCDR2 represented by the amino acid sequence of SEQ ID NO. 34, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 65, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

38. In Paragraph 37, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 126; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

156.

39. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 14, HCDR2 represented by the amino acid sequence of SEQ ID NO. 35, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

98.

40. In Paragraph 39, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 127; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

157.

41. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 15, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 66, LCDR2 represented by the amino acid sequence of SEQ ID NO. 81, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

100.

42. In Paragraph 41, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 128; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

158.

43. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 67, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

101.

44. In Paragraph 43, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 129; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

159.

45. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 12, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 68, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

102.

46. ​​In Paragraph 45, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 130; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

160.

47. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 8, HCDR2 represented by the amino acid sequence of SEQ ID NO. 27, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

103.

48. In Paragraph 47, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 131; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

161.

49. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 16, HCDR2 represented by the amino acid sequence of SEQ ID NO. 36, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 69, LCDR2 represented by the amino acid sequence of SEQ ID NO. 75, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

104.

50. In Paragraph 49, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 132; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

162.

51. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 17, HCDR2 represented by the amino acid sequence of SEQ ID NO. 37, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 70, LCDR2 represented by the amino acid sequence of SEQ ID NO. 84, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

105.

52. In Paragraph 51, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 133; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

163.

53. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 18, HCDR2 represented by the amino acid sequence of SEQ ID NO. 38, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

106.

54. In Paragraph 53, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 134; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

164.

55. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 19, HCDR2 represented by the amino acid sequence of SEQ ID NO. 39, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 71, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

107.

56. In Paragraph 55, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 135; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

165.

57. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 20, HCDR2 represented by the amino acid sequence of SEQ ID NO. 40, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

108.

58. In Paragraph 57, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 136; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

166.

59. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 21, HCDR2 represented by the amino acid sequence of SEQ ID NO. 41, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 52; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 82, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

108.

60. In Paragraph 59, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 137; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

167.

61. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

62. In Paragraph 61, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 138; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

168.

63. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 58, LCDR2 represented by the amino acid sequence of SEQ ID NO. 83, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

217.

64. In Paragraph 63, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 138; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

221.

65. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

66. In Paragraph 65, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 218; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

168.

67. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 215, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

68. In Paragraph 67, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 219; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

168.

69. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 216, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

97.

70. In Paragraph 69, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 220; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

168.

71. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 222, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

228.

72. In Paragraph 71, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 231; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

235.

73. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 223, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 226, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

229.

74. In Paragraph 73, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 232; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

236.

75. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 224, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

230.

76. In Paragraph 75, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 233; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

237.

77. A heavy chain variable region comprising HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46; and An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, comprising a light chain variable region comprising LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

228.

78. In Paragraph 77, The above heavy chain variable region comprises the amino acid sequence of SEQ ID NO. 234; An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO.

238.

79. A polynucleotide encoding the antibody of any one of claims 1 to 78 or the antigen-binding fragment thereof.

80. A vector comprising the polynucleotide of claim 79.

81. Cells transformed with the vector of paragraph 80. 82.i) a step of culturing the transformed cells of claim 81; and ii) a step of obtaining an antibody or an antigen-binding fragment thereof from a culture medium of the cell; a method for preparing an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1.

83. A pharmaceutical composition for the prevention or treatment of cancer comprising, as an active ingredient, an antibody that specifically binds to lipocotin 1 according to any one of claims 1 to 78 or an antigen-binding fragment thereof.

84. In Paragraph 83, A pharmaceutical composition for the prevention or treatment of cancer, wherein the cancer is any one selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, and lymphoma.

85. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 78; and an anticancer agent bound thereto, comprising an antibody-drug conjugate.

86. In Paragraph 85, The antibody or its antigen-binding fragment; and the drug bound thereto are an antibody-drug conjugate formed by being bound through a linker.

87. In Paragraph 86, Antibody-drug conjugate having the structure of structural formula I below: <Structural Formula I> Ab-[LD]n At this time, in the above structural formula I, Ab is an antibody of any one of claims 1 to 78 or an antigen-binding fragment thereof, and L is a linker or direct link, D is an anticancer drug, and n is an integer from 1 to 10.

88. In Paragraph 87, The above linker is an antibody-drug conjugate in which the linker is a non-cleavage type linker or a cleavage type linker.

89. In Paragraph 88, The above-mentioned non-cleavage type linker is a maleimide linker, an antibody-drug conjugate.

90. In Paragraph 88, The above-mentioned cleavage linker is a chemical cleavage linker or an enzyme cleavage linker, an antibody-drug conjugate.

91. In Paragraph 90, An antibody-drug conjugate in which the enzyme-cleaving linker is a peptide-based linker or a specific enzyme-based linker.

92. In Paragraph 90, An antibody-drug conjugate, wherein the above-mentioned cleavage linker is one or more selected from the group consisting of a hydrazone linker, an ester linker, a disulfide linker, a valine-citruline linker, a valine-alanine linker, a phenylalanine-glycine linker, a β-galactoside linker, a β-glucuronide linker, and a phosphodiester linker.

93. In Paragraph 87, The above anticancer agent is an antibody-drug conjugate that is a cytotoxic agent.

94. In Paragraph 93, The above-mentioned cytotoxic agent is an antibody-drug conjugate selected from the group consisting of cytotoxic agents of the Maytansinoid class, Auristatin class, and Calicheamicin class.

95. In Paragraph 93, The above-mentioned cytotoxic agent is an antibody-drug conjugate selected from the group consisting of Ansamitocin, Mertansine, Ravtansine, Monomethyl auristatin E (MMAE), Monomethyl auristatin F (MMAF), and Calicheamicin γ1.

96. In Paragraph 87, The above antibody-drug conjugate is an antibody-drug conjugate represented by the following chemical formula VII: Chemical Formula VII In the above chemical formula, X is NR 1 , O, S or Se and, R 1 H or C 1-8 It is hydrocarbil, and The mAb is an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1 of any one of claims 1 to 78, and n is an integer from 1 to 10.

97. A pharmaceutical composition for the prevention or treatment of cancer comprising an antibody-drug conjugate of any one of claims 85 to 96 as an active ingredient.

98. In Paragraph 97, A pharmaceutical composition for the prevention or treatment of cancer, characterized in that the above cancer is overexpressed with lipocotin-1.

99. In Paragraph 98, A pharmaceutical composition for the prevention or treatment of cancer, wherein the above cancer is any one selected from the group consisting of stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, and lymphoma.

100. An antibody or its antigen-binding fragment that specifically binds to the region from the 27th amino acid to the 185th amino acid of the amino acid sequence of the lipocotin 1 protein represented by sequence number 206.

101. In Paragraph 100, The antibody or its antigen-binding fragment is an antibody or its antigen-binding fragment that specifically binds to the region from the 136th to the 162nd amino acid of the amino acid sequence of the lipocotin 1 protein represented by SEQ ID NO.

206.

102. In Paragraph 100, The above antibody or its antigen-binding fragment is an antibody or its antigen-binding fragment that competes for binding to Lipocotin 1 protein with one or more of the following antibodies or its antigen-binding fragments: An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 30, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 97; An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 222, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 228; An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 223, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 226, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 229; An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 224, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 72, LCDR2 represented by the amino acid sequence of SEQ ID NO. 78, and LCDR3 represented by the amino acid sequence of SEQ ID NO. 230; or An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region including HCDR1 represented by the amino acid sequence of SEQ ID NO. 22, HCDR2 represented by the amino acid sequence of SEQ ID NO. 214, and HCDR3 represented by the amino acid sequence of SEQ ID NO. 46, and a light chain variable region including LCDR1 represented by the amino acid sequence of SEQ ID NO. 225, LCDR2 represented by the amino acid sequence of SEQ ID NO. 227, and LCDR3 represented by the amino acid sequence of SEQ ID NO.

228.

103. An antibody or antigen-binding fragment thereof that specifically binds to lipocotin 1 of any one of claims 1 to 78; or the use of the antibody-drug conjugate of claim 85 for the prevention or treatment of cancer.

104. A method for preventing or treating cancer comprising the step of administering to an individual an antibody or an antigen-binding fragment thereof that specifically binds to lipocotin 1 according to any one of claims 1 to 78; or an antibody-drug conjugate according to claim 85.

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