Ph-dependent Anti-sulfated glycosaminoglycan antibody and antibody-drug conjugate

Abstract

Provided is an antibody that exhibits pH dependence and specifically binds to a sulfated glycosaminoglycan (sGAG).　An antibody specifically binding to a sulfated glycosaminoglycan that is at least one selected from the group consisting of the following (1) to (3):　(1) an antibody that contains a heavy chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 1, CDR2 comprising an amino acid sequence represented by SEQ ID NO: 2 and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 3, and a light chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 4, CDR2 comprising an amino acid sequence represented by LGS and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 6; (2) an antibody that contains a heavy chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 7, CDR2 comprising an amino acid sequence represented by SEQ ID NO: 8 and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 9, and a light chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 10, CDR2 comprising an amino acid sequence represented by AAS and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 12; and (3) an antibody that contains a heavy chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 13, CDR2 comprising an amino acid sequence represented by SEQ ID NO: 14 and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 15, and a light chain having CDR1 comprising an amino acid sequence represented by SEQ ID NO: 16, CDR2 comprising an amino acid sequence represented by WAS and CDR3 comprising an amino acid sequence represented by SEQ ID NO: 18.

Description

pH-dependent anti-sulfated glycosaminoglycan antibodies and antibody-drug conjugates

[0001] The present invention relates to an antibody that specifically binds to sulfated glycosaminoglycan (hereinafter, referred to as sGAG), and an antibody-drug conjugate (ADC) comprising the antibody.

[0002] Glycosaminoglycans are long-chain polysaccharides with repeating units of disaccharides of amino sugars and uronic acid or galactose, and are classified according to the type of constituent sugar, the degree and site of sulfation, and the linkage mode of the constituent sugars. Representative examples include heparan sulfate, heparin, chondroitin sulfate, dermatan sulfate, keratan sulfate, and hyaluronic acid, and they vary in chain length and degree of sulfation. On the cell surface, glycosaminoglycans exist as proteoglycans bound to core proteins and play an important role as receptors for growth factors and other factors.

[0003] The sugar components, degree of sulfation, and chain length of cell surface glycosaminoglycans are known to differ significantly between tumor tissues and normal tissues, and several reports have suggested that glycosaminoglycans can be therapeutic targets for tumors (Non-Patent Documents 1 to 3). A drug conjugate consisting of a cytotoxic compound conjugated to var2csa derived from malaria, which binds to tumor-specific sulfated glycosaminoglycans rich in chondroitin sulfate, has been shown to exhibit antitumor effects in in vivo models (Non-Patent Document 3). While the effectiveness of sulfated glycosaminoglycans as therapeutic targets has been suggested, the tumor-specific sugar chain structure and targeting are not fully understood.

[0004] Antibody-drug conjugates (ADCs), which combine a cytotoxic drug with an antibody that binds to an antigen expressed on the cell surface and can be internalized into the cell, are expected to selectively kill cancer cells because they can deliver drugs selectively to target-expressing cells (see Non-Patent Documents 4 and 5). Repertoire analysis and bioinformatics analysis of tumor-infiltrating B cells in Diffuse-type gastric carcinoma (DGC) have suggested that sulfated glycosaminoglycans are tumor-specific B cell antigens. Anti-sulfated glycosaminoglycan antibodies produced by tumor-infiltrating B cells and their antibody-drug conjugates have been expected to be effective in cancer treatment, but their tumor accumulation and antitumor effects upon administration have not been elucidated (Patent Document 1 and Non-Patent Document 6). Furthermore, since sulfated glycosaminoglycans are abundant in normal tissues, the development of drugs that recognize tumor-specific sugar chain structures or activate tumor-specifically has been desired from a safety standpoint.

[0005] WO2018 / 235855

[0006] Int J Mol Sci. (2020) 21(17), 5983. Biomolecules. (2021) 11(3), 395. Cancer Cell. (2015) 28(4), 500-514. Bioconjugate Chem. (2010) 21, 5-13. Current Opinion in Chemical Biology (2010) 14, 529-537. Cell Rep. (2017) 20(5), 1073-1087.

[0007] The problem to be solved by the present invention is to provide an antibody that specifically binds to sulfated glycosaminoglycan (sGAG) that exhibits pH dependence, and a further problem to be solved by the present invention is to provide an antibody-drug conjugate (ADC) containing the antibody that is useful as an antitumor agent.

[0008] The present invention provides an antibody that binds to sGAG in a pH-dependent manner and an antibody-drug conjugate (ADC) comprising the antibody.

[0009] Therefore, the present invention has the following features: [1] An antibody that specifically binds to a sulfated glycosaminoglycan, which is at least one selected from the group consisting of (1) to (3) below: (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6; (2) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 10, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: AAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 12; and (3) An antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, CDR2 consisting of the amino acid sequence shown in WAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18.[2] The antibody of [1], which is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 20 and having binding activity to sulfated glycosaminoglycans, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, or a light chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 22 and having binding activity to sulfated glycosaminoglycans; (2) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 24 and having binding activity to sulfated glycosaminoglycans, and (3) an antibody comprising a light chain comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26, or a light chain comprising a variable region having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 26 and having binding activity to sulfated glycosaminoglycans; (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; An antibody comprising a light chain having a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain having a variable region that has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans.[3] An antibody according to [1] or [2], which is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 36 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 38 and having binding activity to sulfated glycosaminoglycans; (2) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 40 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 42 and having binding activity to sulfated glycosaminoglycans; (3) An antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 44, or a heavy chain having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 44 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 46, or a light chain having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 46 and having binding activity to sulfated glycosaminoglycans; and (4) an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 48, or a heavy chain having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 50, or a light chain having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 50 and having binding activity to sulfated glycosaminoglycans. [4] The antibody of any of [1] to [3], which is a human antibody. [5] The antibody of any of [1] to [4], whose subclass is an IgG1 antibody. [6] The antibody according to any one of [1] to [5], which binds with high affinity in an acidic pH range.

[0010] [7] An antibody-drug conjugate (ADC) represented by the following general formula 1: Ab-(L-D)p (1) [wherein Ab is an antibody that specifically binds to a sulfated glycosaminoglycan, and is at least one selected from the group consisting of the following (1) to (3): (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6. (2) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 10, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: AAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 12. (3) An antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, CDR2 consisting of the amino acid sequence shown in WAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18. L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), or MC-CL2; D is monomethyl auristatin, maytansinoid, or camptothecin derivative; and p is an integer selected from 1 to 12.[8] The antibody-drug conjugate (ADC) of [7], wherein Ab is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 20 and having binding activity to sulfated glycosaminoglycans, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, or a light chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 22 and having binding activity to sulfated glycosaminoglycans; (2) a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 24 and having binding activity to sulfated glycosaminoglycans, and (3) an antibody comprising a light chain comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26, or a light chain comprising a variable region having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 26 and having binding activity to sulfated glycosaminoglycans; (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence set forth in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; An antibody comprising a light chain having a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain having a variable region that has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans.[9] The antibody-drug conjugate (ADC) of [7] or [8], wherein Ab is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 36 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 38 and having binding activity to sulfated glycosaminoglycans; (2) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 40 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 42 and having binding activity to sulfated glycosaminoglycans; (3) An antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 44 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 46 and having binding activity to sulfated glycosaminoglycans; and (4) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 50 and having binding activity to sulfated glycosaminoglycans.

[10] The antibody-drug conjugate (ADC) of any of [7] to [9], wherein the Ab comprises a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6.

[11] The antibody-drug conjugate (ADC) of any of [7] to

[10] , wherein the Ab comprises a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22.

[12] The antibody-drug conjugate (ADC) of any of [7] to

[11] , wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38.

[13] An antibody-drug conjugate (ADC) according to any one of [7] to

[10] , wherein Ab is an antibody comprising a heavy chain having a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain having a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34.

[14] An antibody-drug conjugate (ADC) according to any one of [7] to

[10] , wherein Ab is an antibody comprising a heavy chain having the amino acid sequence shown in SEQ ID NO: 48, and a light chain having the amino acid sequence shown in SEQ ID NO: 50.

[15] An antibody-drug conjugate (ADC) according to any one of [7] to

[14] , wherein Ab is an IgG1 antibody.

[16] An antibody-drug conjugate (ADC) according to any one of [7] to

[15] , wherein Ab is a human antibody.

[17] An antibody-drug conjugate (ADC) according to any one of [7] to

[16] , wherein Ab is an antibody that binds with higher affinity in an acidic pH range than in a neutral pH range.

[18] The antibody-drug conjugate (ADC) of any of [7] to

[17] , wherein L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), or MC-CL2.

[19] The antibody-drug conjugate (ADC) of any of [7] to

[18] , wherein D is MMAE, MMAF, DM1, DM4, Dx8951, Dxd, or SN-38.

[20] The antibody-drug conjugate (ADC) of any of [7] to

[19] , wherein L-D is SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, MC-GGFG-Dx8591, or MC-CL2-SN-38.

[21] The antibody-drug conjugate (ADC) of any of [7] to

[10] or

[15] to

[20] , wherein Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20 and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, and L-D is MC-Val-Cit-PAB-MMAE.

[22] An antibody-drug conjugate (ADC) according to any one of [7] to

[10] or

[15] to

[20] , wherein Ab is an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and L-D is MC-Val-Cit-PAB-MMAE.

[23] An antibody-drug conjugate (ADC) according to any one of [7] to

[10] or

[15] to

[20] , wherein Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32 and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and L-D is MC-Val-Cit-PAB-MMAE.

[24] An antibody-drug conjugate (ADC) according to any one of [7] to

[10] or

[15] to

[20] , wherein Ab is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:48 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:50, and L-D is MC-Val-Cit-PAB-MMAE.

[25] An antibody-drug conjugate (ADC) according to any one of [7] to

[24] , wherein p is 3 to 8.

[26] A pharmaceutical composition comprising an antibody-drug conjugate (ADC) according to any one of [7] to

[25] and a carrier.

[27] An antitumor agent comprising as an active ingredient an antibody-drug conjugate (ADC) according to any one of [7] to

[25] .

[28] An antitumor agent according to

[27] for treating cancer expressing sulfated glycosaminoglycans.

[29] An antitumor agent according to

[27] for treating gastric cancer, pancreatic cancer, breast cancer, colon cancer, or lung cancer.

[30] A method for treating a tumor, comprising administering to a patient an antibody-drug conjugate (ADC) according to any one of [7] to

[25] .

[31] The method for treating a tumor according to

[30] , wherein the tumor is a cancer expressing sulfated glycosaminoglycans.

[32] The method for treating a tumor according to

[30] , wherein the tumor is gastric cancer, pancreatic cancer, breast cancer, colorectal cancer, or lung cancer.

[33] An antibody-drug conjugate (ADC) according to any one of [7] to

[25] , for use in treating a tumor.

[34] An antibody-drug conjugate (ADC) according to

[33] , wherein the tumor is a cancer expressing sulfated glycosaminoglycans.

[35] An antibody-drug conjugate (ADC) according to

[33] , wherein the tumor is gastric cancer, pancreatic cancer, breast cancer, colorectal cancer, or lung cancer.

[36] Use of an antibody-drug conjugate (ADC) according to any one of [7] to

[25] , for producing an antitumor agent.

[37] Use of

[36] , wherein the tumor is a cancer expressing sulfated glycosaminoglycans.

[38] The use of

[36] , wherein the tumor is gastric cancer, pancreatic cancer, breast cancer, colon cancer, or lung cancer. This specification incorporates the disclosure of Japanese Patent Application No. 2022-057542, from which the present application claims priority.

[0011] The antibody of the present invention specifically binds to sulfated glycosaminoglycans present in tumor tissues in a pH-dependent manner, and is useful not only as a reagent for detecting sGAG, but also as the antibody moiety of antibody-drug conjugates (ADCs). Furthermore, the antibody-drug conjugates (ADCs) of the present invention exhibit excellent antitumor effects with minimal side effects on normal tissues due to the pH dependence of the antibody.

[0012] 1 shows the pH-dependent binding activity of each sGAG antibody. This figure shows that the pH-dependent binding of each sGAG antibody is dependent on the target antigen due to the inhibition of sGAG formation. This figure shows target expression in normal cell lines and cancer cell lines using antibody 1. This figure shows the combination of antibodies and linker-payload for each ADC (part 1). This figure shows the combination of antibodies and linker-payload for each ADC (part 2). This figure shows the 50% effective concentration for the NUGC-4 cell line, when the maximum binding activity at pH 6.0 is taken as 100%, for each antibody and its ADC. This figure shows the stability of the drug linker in mouse and human plasma for each ADC. This figure shows the cytocidal effect of antibodies 1 to 3 and their ADCs 1 to 3 against NUGC-3. This figure shows the cytocidal effect of ADCs 4 to 10 against NUGC-4 and Capan-1. This figure shows the antitumor effect of ADC 6 against a mouse model subcutaneously implanted with the human pancreatic cancer cell line BxPC-3.

[0023] Figure 1 shows the antitumor effect of ADC6 on a mouse model in which the human scirrhous gastric cancer cell line NUGC-4 is subcutaneously implanted. Figure 2 shows the sequences of SEQ ID NOs: 1 to 4, 6 to 10. Figure 3 shows the sequences of SEQ ID NOs: 12 to 16, 18 to 20. Figure 4 shows the sequences of SEQ ID NOs: 21 to 26. Figure 5 shows the sequences of SEQ ID NOs: 27 to 32. Figure 6 shows the sequences of SEQ ID NOs: 33 to 36. Figure 7 shows the sequences of SEQ ID NOs: 37 to 39. Figure 8 shows the sequences of SEQ ID NOs: 40 to 42. Figure 9 shows the sequences of SEQ ID NOs: 43 to 45. Figure 10 shows the sequences of SEQ ID NOs: 46 to 48. Figure 11 shows the sequences of SEQ ID NOs: 49 and 50.

[0013] The present invention is described in detail below: The present invention provides an antibody against a sulfated glycosaminoglycan and an antibody-drug conjugate (ADC) of the antibody for cancer treatment.

[0014] <Antibody> The present invention provides an antibody that binds to sulfated glycosaminoglycans with high affinity in the acidic pH range.

[0015] The antibodies of the present invention are characterized by binding to sulfated glycosaminoglycans with high affinity in the acidic pH range, and are therefore useful as reagents for the detection, diagnosis, isolation, etc. of sulfated glycosaminoglycans under acidic pH conditions. Furthermore, due to their tumor tissue selectivity based on their pH dependence, they are useful as antibody moieties in antibody-drug conjugates (ADCs) with antitumor activity.

[0016] In the present invention, "sulfated glycosaminoglycan" refers to a glycosaminoglycan in which some of the constituent disaccharide units have been sulfated, regardless of the type of the constituent disaccharide units (hyaluronic acid, chondroitin sulfate, dermatan sulfate, keratan sulfate, heparan sulfate, heparin, etc.), the degree of sulfation, or the sulfation site.

[0017] In the present invention, the phrase "binding with high affinity in an acidic pH range" means binding with higher affinity in an acidic pH range than in a neutral pH range. Specifically, this means that the affinity of the antibody of the present invention when binding to sGAG is at least four times, preferably at least eight times, and more preferably at least ten times higher in an acidic pH range (5.0 to 6.8, preferably 5.5 to 6.4, more preferably 6.0) than in the pH range in blood (7.4). The affinity of the antibody of the present invention when binding to sGAG can be measured by known techniques, for example, as the Alexa Fluor 488 signal (MFI: Mean Fluorescence Intensity) by flow cytometry as described in Example 2 below. Antibodies whose binding affinity varies with pH in this way are called pH-dependent antibodies.

[0018] The antibodies of the present invention also have internalization ability, bind to sulfated glycosaminoglycans in tumor tissues, and are taken up into tumor cells.

[0019] The antibody of the present invention is at least one selected from the group consisting of the following (1) to (3):

[0020] (1) An antibody comprising a heavy chain comprising a CDR1 (complementarity determining region) consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown as LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6.

[0021] (2) An antibody comprising a heavy chain comprising CDR1 having the amino acid sequence shown in SEQ ID NO: 7, CDR2 having the amino acid sequence shown in SEQ ID NO: 8, and CDR3 having the amino acid sequence shown in SEQ ID NO: 9, and a light chain comprising CDR1 having the amino acid sequence shown in SEQ ID NO: 10, CDR2 having the amino acid sequence shown in AAS, and CDR3 having the amino acid sequence shown in SEQ ID NO: 12.

[0022] (3) An antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, CDR2 consisting of the amino acid sequence shown in WAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18.

[0023] Among these, preferred is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6.

[0024] Furthermore, a preferred antibody of the present invention is at least one selected from the group consisting of the following (1) to (4): (1) An antibody comprising a heavy chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 20, and a light chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 22. (2) An antibody comprising a heavy chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 24, and a light chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26. (3) An antibody comprising a heavy chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 28, and a light chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 30. (4) An antibody comprising a heavy chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 32, and a light chain including a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 34.

[0025] Among these, more preferred are: (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22; or (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34; and particularly preferred is (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34.

[0026] The heavy chain variable region includes not only heavy chain variable regions consisting of the amino acid sequence represented by the above SEQ ID NO, but also heavy chain variable regions consisting of an amino acid sequence in which one or several, for example, 1 to 10, preferably 1 to 5, more preferably 1 or 2, and even more preferably 1 amino acid has been deleted, substituted, or added in the amino acid sequence other than the CDRs, and which consist of a protein having the activity of the heavy chain variable region of an antibody, i.e., binding activity to sulfated glycosaminoglycans. The light chain variable region includes not only light chain variable regions consisting of the amino acid sequence represented by the above SEQ ID NO, but also light chain variable regions consisting of an amino acid sequence in which one or several, for example, 1 to 10, preferably 1 to 5, more preferably 1 or 2, and even more preferably 1 amino acid has been deleted, substituted, or added in the amino acid sequence other than the CDRs, and which consist of a protein having the activity of the light chain variable region of an antibody, i.e., binding activity to sulfated glycosaminoglycans. The presence or absence of binding activity to sulfated glycosaminoglycans can be measured by known techniques.

[0027] Such amino acid sequences in which one or several amino acids have been deleted, substituted or added in the amino acid sequence shown by the above SEQ ID NO: include those which have at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence of the portion other than the CDRs in the amino acid sequence shown by the above SEQ ID NO: when calculated using BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information) or the like (for example, default, i.e., initial setting parameters).

[0028] Such a protein having an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in the above SEQ ID NO: is substantially identical to the protein consisting of the amino acid sequence shown in the above SEQ ID NO:.

[0029] Furthermore, a preferred antibody of the present invention is at least one selected from the group consisting of the following (1) to (4):

[0030] (1) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 20, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 6, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 22, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 22;(2) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 24, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence set forth in AAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 12, and comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 26 and has binding activity to sulfated glycosaminoglycans;(3) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 28, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence set forth in WAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 18, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 30, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 30 and has binding activity to sulfated glycosaminoglycans;and (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO:4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO:6, and comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO:34, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:34, and having binding activity to sulfated glycosaminoglycans;

[0031] Among these, more preferred are: (1) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 20, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 22; or(4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, and more preferably(4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, and more preferably (4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; andAn antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans.

[0032] Particularly preferred is an antibody comprising: (4) a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a variable region having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain comprises a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and having binding activity to sulfated glycosaminoglycans.

[0033] Furthermore, a preferred antibody of the present invention is at least one selected from the group consisting of the following (1) to (4): (1) An antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38 (Antibody 1). (2) An antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42 (Antibody 2). (3) An antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46 (Antibody 3). (4) An antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50 (Antibody 4).

[0034] Among these, more preferred are: (1) an antibody (antibody 1) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38; or (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50; and particularly preferred, from the viewpoints of high production efficiency and low aggregation tendency, is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50.

[0035] The heavy chain includes not only a heavy chain consisting of the amino acid sequence shown by the above SEQ ID NO, but also a heavy chain consisting of an amino acid sequence in which one or several, for example, 1 to 10, preferably 1 to 5, more preferably 1 or 2, and even more preferably 1 amino acid has been deleted, substituted, or added in the amino acid sequence in the portion other than the CDRs of the amino acid sequence, and which consists of a protein having antibody heavy chain activity, i.e., binding activity to sulfated glycosaminoglycans. The light chain includes not only a light chain consisting of the amino acid sequence shown by the above SEQ ID NO, but also a light chain consisting of an amino acid sequence in which one or several, for example, 1 to 10, preferably 1 to 5, more preferably 1 or 2, and even more preferably 1 amino acid has been deleted, substituted, or added in the amino acid sequence in the portion other than the CDRs of the amino acid sequence, and which consists of a protein having antibody light chain activity, i.e., binding activity to sulfated glycosaminoglycans. The presence or absence of binding activity to sulfated glycosaminoglycans can be measured by known techniques.

[0036] Such amino acid sequences in which one or several amino acids have been deleted, substituted or added in the amino acid sequence shown by the above SEQ ID NO: include those which have at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence of the portion other than the CDRs in the amino acid sequence shown by the above SEQ ID NO: when calculated using BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information) or the like (for example, default, i.e., initial setting parameters).

[0037] Furthermore, a preferred antibody of the present invention is at least one selected from the group consisting of the following (1) to (4):

[0038] (1) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 36, and having binding activity to sulfated glycosaminoglycans; and An antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain consists of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 38, and having binding activity to sulfated glycosaminoglycans.

[0039] (2) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and having a sequence identity of at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% with the amino acid sequence shown in SEQ ID NO: 40, and having binding activity to sulfated glycosaminoglycans; and An antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence shown in AAS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 12, wherein the light chain consists of the amino acid sequence shown in SEQ ID NO: 42, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 42, and having binding activity to sulfated glycosaminoglycans.

[0040] (3) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and having at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 44, and having binding activity to sulfated glycosaminoglycans; An antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence shown in WAS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18, wherein the light chain consists of the amino acid sequence shown in SEQ ID NO: 46, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 46, and having binding activity to sulfated glycosaminoglycans.

[0041] (4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and having a sequence identity of at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% with the amino acid sequence shown in SEQ ID NO: 48, and having binding activity to sulfated glycosaminoglycans; An antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain consists of the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 50, and having binding activity to sulfated glycosaminoglycans.

[0042] Among these, more preferred are: (1) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 36, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain consists of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 38, and having binding activity to sulfated glycosaminoglycans; or(4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and having a sequence identity of at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% with the amino acid sequence shown in SEQ ID NO: 48, and having binding activity to sulfated glycosaminoglycans; an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and which comprises an amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 50, and which has binding activity to sulfated glycosaminoglycans, and more preferably(4) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and having a sequence identity of at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% with the amino acid sequence shown in SEQ ID NO: 48, and having binding activity to sulfated glycosaminoglycans; The antibody comprises a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and the light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 50, and having binding activity to sulfated glycosaminoglycans.

[0043] More preferably, (4) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 48, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans; and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain comprises the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 50 and having binding activity to sulfated glycosaminoglycans.

[0044] Particularly preferred is an antibody comprising: (4) a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 48, or a heavy chain having at least 98% sequence identity with the amino acid sequence shown in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans; and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, wherein the light chain comprises the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 98% sequence identity with the amino acid sequence shown in SEQ ID NO: 50 and having binding activity to sulfated glycosaminoglycans.

[0045] Such a protein having an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in the above SEQ ID NO: is substantially identical to the protein consisting of the amino acid sequence shown in the above SEQ ID NO:.

[0046] The sequence information of "Antibody 1," "Antibody 2," "Antibody 3," and "Antibody 4," which are preferred antibodies in the present invention, is shown in Table 1.

[0047]

[0048] The form of the antibody of the present invention is not particularly limited as long as it is a molecule having a pair of heavy and light chains and capable of binding to an antigen. For example, in addition to immunoglobulins, antigen-binding fragments or derivatives thereof (Fab fragments, F(ab')) having all or part of the functions of a complete antibody can be used. ２ Fragments, single chain antibody fragments (scFv, variable region (Fv), dF(ab') ２ The antibody may also include Fab'iabodies, which are monovalent fragments of the variable region of an antibody prepared by treating the antibody under reducing conditions, and bispecific antibodies.

[0049] The origin of the antibodies of the present invention is not particularly limited, and examples include humans, rats, mice, rabbits, hamsters, guinea pigs, horses, monkeys, dogs, pigs, cattle, goats, and sheep. The antibodies of the present invention are preferably recombinant monoclonal antibodies. When derived from a species other than humans, they can be chimerized or humanized using well-known techniques. Chimeric antibodies are antibodies having human constant regions, and humanized antibodies are antibodies in which the CDRs (complementarity-determining regions) of a human antibody are grafted with the CDRs of an anti-sulfated glycosaminoglycan antibody from a non-human animal (CDR-grafted antibodies). Furthermore, the antibodies of the present invention include human antibodies, which are the expression products of antibody genes derived from humans. In the present invention, preferred are human antibodies, humanized antibodies, and chimeric antibodies, and more preferably human antibodies. Furthermore, when the antibodies of the present invention are human antibodies, their subtypes are not particularly limited, and IgG1, IgG2, IgG3, and IgG4 are preferred, with IgG1 being more preferred. Examples of heavy chain constant regions include Cγ1, Cγ2, Cγ3, and Cγ4, and examples of light chain constant regions include Cκ and Cλ.

[0050] The antibody of the present invention can be produced using various known methods, and the production method is not particularly limited. Such known methods include the hybridoma method and the phage display method.

[0051] The antibodies of the present invention can be modified by known recombinant means, provided that the effects of the present invention are not impaired. For example, at least one amino acid in the antibody constant region can be substituted with a different residue. Such changes are made to reduce undesirable activities, such as complement-dependent cytotoxicity.

[0052] The antibodies of the present invention can be produced as recombinant antibodies. Specifically, DNA encoding each region of the antibody (DNA encoding the heavy chain variable region, light chain variable region, heavy chain variable region, and heavy chain constant region) can be inserted into an expression vector, host cells can be transformed with the expression vector, and the cells can be cultured to produce the antibody. In this case, the DNA encoding each region may be operably linked to elements such as a promoter, polyadenylation signal, or enhancer. "Operatively linked" refers to linking each element so that it functions. The DNA used to insert each region into the vector may contain a signal peptide that promotes antibody secretion from host cells. The mature protein can be obtained by removing the signal peptide after antibody production. Known plasmids, phages, and the like can be used as expression vectors. Prokaryotic cells such as Escherichia coli and Bacillus subtilis, and eukaryotic cells such as yeast and animal cells can be used as host cells. Among these, eukaryotic cells in which a sugar chain is bound to the antibody are preferred. Suitable animal cells include Chinese hamster ovary cells (CHO) and HEK293 cells. The expression vector may be introduced into host cells by electroporation, DEAE-dextran transfection, calcium phosphate precipitation, or the like, to transform the host cells. The produced antibody may be purified by known methods, such as methods using various types of chromatography. It is known that antibodies produced in cultured mammalian cells lack a lysine residue at the carboxyl terminus of the heavy chain, and the present invention also includes antibodies in which a lysine residue at the carboxyl terminus of the heavy chain is deleted.

[0053] <Antibody-drug conjugates (ADCs)> The present invention provides antibody-drug conjugates (ADCs) represented by the following general formula 1: Ab-(L-D)p [wherein Ab is an antibody that specifically binds to a sulfated glycosaminoglycan and is at least one selected from the group consisting of the following (1) to (3): (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6.

[0054] (2) An antibody comprising a heavy chain comprising CDR1 having the amino acid sequence shown in SEQ ID NO: 7, CDR2 having the amino acid sequence shown in SEQ ID NO: 8, and CDR3 having the amino acid sequence shown in SEQ ID NO: 9, and a light chain comprising CDR1 having the amino acid sequence shown in SEQ ID NO: 10, CDR2 having the amino acid sequence shown in AAS, and CDR3 having the amino acid sequence shown in SEQ ID NO: 12.

[0055] (3) An antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, CDR2 consisting of the amino acid sequence shown in WAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18.

[0056] L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), or MC-CL2; D is monomethyl auristatin, maytansinoid, or camptothecin derivative; and p is an integer selected from 1 to 12.

[0057] The Ab in the ADC of the present invention is preferably the above-mentioned antibody.

[0058] In the ADCs of the present invention, L is a linker connecting an antibody and a drug, which may or may not contain an enzymatic cleavage site. Examples of linkers include 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB: MC-VC-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), and MC-CL2 (J Med Chem. 2008 November 13; 51(21): 6916-6926). For example, Val-Cit (valine-citrulline) is a peptide linker that combines valine and citrulline, and is cleaved by cysteine ​​protease enzymes such as cathepsin.

[0059] D in the ADC of the present invention is a drug, and examples thereof include anticancer agents such as monomethyl auristatin (MMA), maytansinoids, camptothecin derivatives, etc. Monomethyl auristatins include monomethyl auristatin E (MMAE), monomethyl auristatin D (MMAD), monomethyl auristatin F (MMAF), etc., with monomethyl auristatin E (MMAE) being preferred. Maytansinoids include DM1 (N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-Maytansine), DM4, etc. Camptothecin derivatives include topocathen, irinotecan, Dx-8951, Dx-d, SN-38, etc.

[0060] Preferred combinations of L and D in the ADCs of the present invention include SMCC-maytansinoid, SPDB-maytansinoid, MC-Val-Cit-PAB-monomethylauristatin, MC-Val-Cit-PAB-monomethylauristatin, MC-GGFG-camptothecin derivative, or MC-CL2-camptothecin derivative, more preferably SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, MC-GGFG-Dx8591, or MC-CL2-SN-38, more preferably SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, or MC-GGFG-Dx8591, and even more preferably MC-Val-Cit-PAB-MMAE.

[0061] In the ADC of the present invention, p is an integer selected from 1 to 12, preferably 1 to 10, more preferably 2 to 9, and particularly preferably 3 to 8.

[0062] The ADC of the present invention can be produced by known methods. For example, in an ADC, a drug is bound to an antibody via a linker, and the linker is bound to the antibody via an amino group or cysteine ​​residue of the antibody. Specifically, the linker is bound to the antibody via a Michael addition reaction between a sulfhydryl group of cysteine ​​and maleimide, a disulfide bond formation reaction, or an amide bond formation by condensation of a lysine amino group with a carboxylic acid. In the above general formula 1, p represents the number of drug molecules that can be bound to one antibody molecule. This number of molecules is called the DAR (Drug Antibody Ratio).

[0063] Furthermore, a preferred ADC of the present invention is an antibody that specifically binds to a sulfated glycosaminoglycan, wherein Ab in general formula 1 is at least one selected from the group consisting of the following (1) to (4): (1) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 20, and comprising a variable region that has binding activity to a sulfated glycosaminoglycan; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 6, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 22, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 22;(2) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 24, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence set forth in AAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 12, and comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 26 and has binding activity to sulfated glycosaminoglycans;(3) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 28, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence set forth in WAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 18, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 30, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 30 and has binding activity to sulfated glycosaminoglycans;and (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO:4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO:6, and comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO:34, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:34, and having binding activity to sulfated glycosaminoglycans;

[0064] an ADC in which L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), or MC-CL2; D is monomethyl auristatin, maytansinoid, or camptothecin derivative; and p is an integer selected from 1 to 12.

[0065] More preferably, in general formula 1, Ab is an antibody that specifically binds to a sulfated glycosaminoglycan, and is at least one selected from the group consisting of (1) to (4) below: (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:22; (2) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:24, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:26; (3) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:28, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:30; and (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:34.

[0066] an ADC in which L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyric acid-N-hydroxysuccinimide ester (SPDB), or MC-CL2; D is monomethyl auristatin, maytansinoid, or camptothecin derivative; and p is an integer selected from 1 to 12.

[0067] More preferably, in general formula 1, Ab is an antibody that specifically binds to a sulfated glycosaminoglycan, and is at least one selected from the group consisting of (1) to (4) below: (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:22; (2) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:24, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:26; (3) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:28, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:30; and (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:34.

[0068] L-D is SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, MC-GGFG-Dx8591, or MC-CL2-SN-38, and p is an integer selected from 1 to 12.

[0069] More preferably, in general formula 1, Ab is an antibody that specifically binds to a sulfated glycosaminoglycan, and is at least one selected from the group consisting of (1) to (4) below: (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:22; (2) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:24, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:26; (3) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:28, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:30; and (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO:34.

[0070] LD is SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, or MC-GGFG-Dx8591; and p is an integer selected from 1 to 12.

[0071] More preferably, in general formula 1, Ab is an antibody that specifically binds to a sulfated glycosaminoglycan, and is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38 (antibody 1); (2) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42 (antibody 2); (3) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46 (antibody 3); (4) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50 (antibody 4).

[0072] LD is SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, or MC-GGFG-Dx8591; and p is an integer selected from 1 to 12.

[0073] A preferred ADC of the present invention is an ADC represented by the following general formula 2: [wherein Ab is at least one antibody selected from the group consisting of (1) to (3) below that specifically binds to sulfated glycosaminoglycans, and is: (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6; (2) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 10, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: AAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 12; and (3) An antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 16, CDR2 consisting of the amino acid sequence shown in WAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 18, where p is an integer selected from 1 to 12.

[0074] More preferably, in the formula, Ab is at least one selected from the group consisting of the following (1) to (4): (1) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 20, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 6, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 22, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 22;(2) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 24, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence set forth in AAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 12, and comprising a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 26, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 26 and has binding activity to sulfated glycosaminoglycans;(3) A heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, and a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, or a heavy chain having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 28, and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence set forth in WAS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 18, wherein the light chain comprises a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 30, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 30 and has binding activity to sulfated glycosaminoglycans;and (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, wherein the heavy chain comprises a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32, and having binding activity to sulfated glycosaminoglycans; and An antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence set forth in LGS, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 6, and a variable region consisting of the amino acid sequence set forth in SEQ ID NO: 34, or a light chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 34, and having binding activity to sulfated glycosaminoglycans.

[0075] More preferably, in the formula, Ab is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22. (2) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 26. (3) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 30. (4) an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34. An ADC in which p is an integer selected from 1 to 12,

[0076] Particularly preferred are antibodies in which Ab is at least one selected from the group consisting of the following (1) to (4): (1) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38 (antibody 1); (2) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42 (antibody 2); (3) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46 (antibody 3); (4) an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50 (antibody 4); and p is an integer selected from 1 to 12, and is an ADC.

[0077] A preferred ADC of the present invention is an ADC represented by the following general formula 3: [wherein Ab is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and p is an integer selected from 1 to 12.]

[0078] More preferably, the Ab is (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, wherein p is an integer selected from 1 to 12;

[0079] More preferably, the ADC is an antibody (4) in which Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and p is an integer selected from 1 to 12;

[0080] Particularly preferred is an ADC in which Ab is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, and p is an integer selected from 1 to 12.

[0081] A preferred ADC of the present invention is an ADC represented by the following general formula 4: [wherein Ab is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and p is an integer selected from 1 to 12.]

[0082] More preferably, the Ab is (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, wherein p is an integer selected from 1 to 12;

[0083] More preferably, the ADC is an antibody (4) in which Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and p is an integer selected from 1 to 12;

[0084] Particularly preferred is an ADC in which Ab is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, and p is an integer selected from 1 to 12.

[0085] A preferred ADC of the present invention is an ADC represented by the following general formula 5: [wherein Ab is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and p is an integer selected from 1 to 12.]

[0086] More preferably, the Ab is (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, wherein p is an integer selected from 1 to 12;

[0087] More preferably, the ADC is an antibody (4) in which Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and p is an integer selected from 1 to 12;

[0088] Particularly preferred is an ADC in which Ab is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, and p is an integer selected from 1 to 12.

[0089] A preferred ADC of the present invention is an ADC represented by the following general formula 6: [wherein Ab is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and p is an integer selected from 1 to 12.]

[0090] More preferably, the Ab is (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, wherein p is an integer selected from 1 to 12;

[0091] More preferably, the ADC is an antibody (4) in which Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and p is an integer selected from 1 to 12;

[0092] Particularly preferred is an ADC in which Ab is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, and p is an integer selected from 1 to 12.

[0093] A preferred ADC of the present invention is an ADC represented by the following general formula 7: [wherein Ab is (1) an antibody comprising a heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and p is an integer selected from 1 to 12.]

[0094] More preferably, the Ab is (4) a heavy chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, a CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain comprising a variable region having at least 85%, preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, and particularly preferably at least 99% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans; and an antibody comprising a light chain comprising a CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, a CDR2 consisting of the amino acid sequence shown in LGS, and a CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6, and comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain comprising a variable region which has at least 85%, preferably at least 90%, more preferably at least 91%, even more preferably at least 92%, even more preferably at least 93%, even more preferably at least 94%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and particularly preferably at least 99% sequence identity to the amino acid sequence shown in SEQ ID NO: 34 and has binding activity to sulfated glycosaminoglycans, wherein p is an integer selected from 1 to 12;

[0095] More preferably, the ADC is an antibody (4) in which Ab is an antibody comprising a heavy chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain comprising a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, and p is an integer selected from 1 to 12;

[0096] Particularly preferred is an ADC in which Ab is (4) an antibody (antibody 4) comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, and p is an integer selected from 1 to 12.

[0097] The linker-drug structure and linker structure used in the ADC of the present invention are shown below. In the diagram of the linker, the linker is bound to the antibody on the left side via the dashed line, and to the drug on the right side.

[0098] ADC1~4 (SMCC-DM1)

[0099] SMCC

[0100] DM1

[0101] ADC5 (SPDB-DM4)

[0102] SPDB

[0103] DM4

[0104] ADC6 (MC-Val-Cit-PAB-MMAE)

[0105] MC-Val-Cit-PAB

[0106] MMAE

[0107] ADC7 (MC-GGFG-Dxd)

[0108] MC-GGFG

[0109] Dxd

[0110] ADC8 (MC-GGFG-Dx8951)

[0111] MC-GGFG

[0112] DX8951

[0113] ADC9 (MC-Val-Cit-PAB-MMAF)

[0114] MC-Val-Cit-PAB

[0115] MMAF

[0116] ADC10 (MC-CL2-SN-38)

[0117] MC-CL2

[0118] SN-38

[0119] <Anticancer Agent> The present invention also includes an anticancer agent containing the above-mentioned ADC as an active ingredient.

[0120] The cancer on which the antibody-drug conjugate (ADC) of the present invention exerts an antitumor effect is not particularly limited as long as it is a cancer that expresses sGAG, and specific examples include gastric cancer, pancreatic cancer, colon cancer, liver cancer, lung cancer, breast cancer, skin cancer, oral cancer, esophageal cancer, bile duct cancer, uterine cancer, cervical cancer, ovarian cancer, renal cell carcinoma, urothelial cancer, and prostate cancer. Preferably, it is a solid cancer, more preferably, breast cancer, ovarian cancer, uterine cancer, cervical cancer, neuroblastoma, glioma, glioblastoma, bladder cancer, melanoma, colon cancer, colorectal cancer, gastric cancer, lung cancer, pancreatic cancer, prostate cancer, liver cancer, nasopharyngeal cancer, renal cancer, multiple myeloma, oral squamous cell carcinoma, esophageal adenocarcinoma, esophageal squamous cell carcinoma, peripheral nerve sheath tumor (Int. J. Mol. Sci. 2020, 21(17), 5983), and particularly preferably, gastric cancer, pancreatic cancer, breast cancer, colorectal cancer, or lung cancer.

[0121] The ADC of the present invention is taken up into the above-mentioned cancer cells and exerts a therapeutic effect.

[0122] Although some sGAG is expressed in normal tissues as well, the antibodies used in the ADCs of the present invention are pH-dependent and therefore do not bind to sGAG in normal tissues, which have a neutral pH environment. As a result, they are unlikely to be taken up by cells in normal tissues and are unlikely to cause side effects on normal tissues.

[0123] The administration route of anticancer agents containing the ADCs of the present invention is not limited, and they can be administered orally, parenterally, or the like. Parenteral administration includes intravenous administration, intramuscular administration, subcutaneous administration, and the like. Furthermore, they may be administered transmucosally (e.g., sublingual or buccal administration), transdermal administration, or rectally. Administration may be systemic or local administration to cancer tissue. Examples of dosage forms for the formulation include tablets, capsules, pills, powders, granules, and injections. Oral formulations such as tablets, capsules, pills, powders, and granules can be prepared using additives such as excipients such as glucose, sucrose, lactose, and mannitol; disintegrants such as starch and sodium alginate; lubricants such as magnesium stearate and talc; binders such as polyvinyl alcohol, gelatin, and hydroxypropyl cellulose; surfactants such as fatty acid esters; and plasticizers such as glycerin. The injection preparation may contain additives such as water, sugars such as sucrose, sorbitol, xylose, trehalose, and fructose; sugar alcohols such as sorbitol, mannitol, and xylitol; buffers such as phosphate buffer, citrate buffer, and glutamate buffer; and surfactants such as fatty acid esters.

[0124] The dosage of ADC administered to a patient in need of treatment varies depending on the age, sex, severity, etc. of the patient, and can be determined by the attending physician. For example, ADC may be administered at 0.05 to 10 mg / kg body weight, preferably 0.1 to 2 mg / kg body weight per dose. Alternatively, ADC may be administered at a drug equivalent of MMAE or the like at 0.001 nM to 1000 nM, preferably 0.001 nM to 100 nM per dose.

[0125] The dose may be administered once or in divided doses, such as two, three, four or more times per day, at appropriate intervals over a period ranging from one day to several years.

[0126] Anticancer agents containing the ADC of the present invention as an active ingredient can be used in combination with other anticancer agents or other therapies, such as surgery, radiation therapy, and chemotherapy.

[0127] The present invention further encompasses a method for treating tumors, comprising administering the ADC to a patient in need of treatment. The tumors to be treated are as described above, and the administration method is also as described above.

[0128] The present invention further includes the above-mentioned ADC for use in treating a tumor. The tumor to be treated is as described above, and the administration method is also as described above.

[0129] The present invention further encompasses use of the ADC for the manufacture of an antitumor agent. The tumor to be treated is as described above, and the administration method is also as described above.

[0130] The present invention will now be described in detail with reference to examples, but the present invention is not limited to these examples.

[0131] Example 1: Production of pH-dependent anti-sGAG monoclonal antibodies (mABs) using recombinant DNA methods. cDNA constructs were prepared from the heavy and light chain genes encoding antibodies 1 to 4 listed in Table 2. Each of the cDNA constructs encoding the heavy and light chains was integrated into pcDNA3 plasmid (Life Technologies), and the respective clonal combinations were transfected into HEK293 cells to forcibly express the heavy and light chains. The culture supernatant was filtered through a 0.22 μm filter, followed by Protein A purification and gel filtration purification to obtain pH-dependent anti-sGAG antibodies.

[0132]

[0133] [Example 2: pH-dependent binding of anti-sGAG antibody to cancer cells] Gastric cancer cell line NUGC-3 (HSRRB) was washed with PBS and then collected with a scraper. The cell number was measured and 1 × 10 ６The cells were collected into a 96-well plate at 100 cells / well. The anti-sGAG antibody obtained in Example 1 and Human IgG Isotype Control (Invitrogen) were diluted to a final concentration of 15 μg / mL using 1% BSA / PBS (pH 8.0-5.0) adjusted to each pH. The diluted solution was added to NUGC-3 and allowed to stand at room temperature for 30 minutes. After the sGAG antibody reaction, the cells were washed with PBS (pH 8.0-5.0), and anti-Human IgG-Alexa Fluor 488 (Invitrogen) dissolved in 1% BSA / PBS (pH 8.0-5.0) was added. After the secondary antibody reaction, the cells were washed with PBS (pH 8.0 to 5.0), and the Alexa Fluor 488 signal (MFI: Mean Fluorescence Intensity) was detected using a flow cytometer (CytoFLEX S, Beckman Coulter). As a result, a pH-dependent increase in binding ability was observed in a low pH environment of pH 6.8 to 5.5 for all sGAG antibodies evaluated (Figure 1).

[0134] It has been shown that PNP-xyl reduces the amount of GAG on the cell surface by competitive inhibition (Biochemical Pharmacology, 1991, 42, (10), pp. 1987-1995, Oncotarget, 2017, 8, (40), pp. 66960-66974). The pH-dependent binding of sGAG antibodies was significantly inhibited by treating cultured cells with PNP-Xyl (4 mM, 48 hours), indicating that the pH-dependent binding to cells is not due to a nonspecific effect (Figure 2).

[0135] Example 3: Evaluation of binding ability of sGAG antibody to multiple cancer cell lines Using the same method as in Example 2, the binding ability of antibody 1 to multiple cancer cell lines was evaluated. The binding ability of the isotype control antibody and Antibody 1 at pH 6.0 was evaluated in gastric cancer cell lines (NCI-N87 (ATCC), NUGC-3 (HSRRB)), pancreatic cancer cell lines (PANC-1 (Dainippon Pharmaceutical Co., Ltd.), AsPC-1 (JCRB), KP-4 (HSRRB), SUIT2 (JCRB)), breast cancer cell line (MDA-MB-231 (ATCC)), colon cancer cell lines (SK-CO-1 (ATCC), HCT116 (ATCC)), and lung cancer cell lines (NCI-H460 (ATCC), DMS273 (ECACC), A549 (Dainippon Pharmaceutical Co., Ltd.)). Expression of the Antibody 1 target antigen was confirmed in all cancer cell lines evaluated, demonstrating its therapeutic utility against multiple cancer types (Figure 3).

[0136] Example 4: Antibody-drug conjugation of anti-sGAG antibody The ADC of the present invention can be prepared by reacting an antibody having an amino group and a sulfhydryl group with a drug linker to be added, using the method described below. The combinations of drug linkers conjugated to the antibody for ADCs obtained using anti-sGAG antibodies are shown in Figure 4. The produced ADC can be identified by concentrating, buffer exchanging, purifying, measuring the antibody concentration, and measuring the average number of drugs bound per antibody molecule using the following common procedures.

[0137] Common Procedure A: Concentration of antibody or ADC solution The antibody or ADC solution was placed in an Amicon Ultra (100,000 MWCO, Millipore Co.) container, and the antibody or ADC solution was concentrated by centrifugation using a centrifuge (himac CF5RE, HITACHI) (centrifugation at 2000 G to 15000 G for 5 to 20 minutes). Common Procedure B: Measurement of antibody concentration The antibody concentration was measured using a UV measurement device (Nanodrop 8000, Thermo Fisher Scientific Inc.) according to the method specified by the manufacturer. Common Procedure C: Buffer Exchange and Purification of Antibody / ADC A Zeba™ Spin Desalting Column (7K MWCO to 40K MWCO, Cat. 89890, Thermo Scientific) was equilibrated with D-PBS (Nacalai Tesque Inc.) according to the method specified by the manufacturer. After loading the antibody / ADC solution, it was eluted with the amount of D-PBS specified by the manufacturer, and the concentration was measured by common procedure B. Common Procedure D-1: Measurement of antibody concentration in ADC and average number of drugs bound per antibody molecule The drug-antibody ratio of DM1 bound to the antibody can be calculated by measuring the UV absorbance of the ADC solution at two wavelengths, 280 nm and 252 nm, and then performing the following calculation.

[0138]

[0139] Here, R is the ADC A252 / A280 = Absorbance ratio, is the molar absorption coefficient of the antibody used at 252 nm or 280 nm, indicates the molar absorption coefficient of DM1 at 252 nm or 280 nm.

[0140] teeth, was used.

[0141] was estimated from the amino acid sequence of each antibody by a known calculation method (Protein Science, 1995, 4, pp. 2411-2423). Antibody 1 was 214,460 M －１ cm －１ , antibodies 2 and 3 are 207360M－１ cm －１ was used.

[0142] The actual average value of each antibody was used.

[0143] Common Procedure D-2: Measurement of the average number of drugs bound per antibody molecule in ADC. The drug-antibody ratio was analyzed by native size-exclusion chromatography mass spectrometry (SEC-MS). SEC-MS analysis allows for the measurement of the drug-antibody ratio based on the mass change caused by the binding of the drug linker to the antibody. 2 mg / mL of ADC was injected into an Acquity LC Xevo G2-S Q Tof MS (Waters Corporation), and the ADC was eluted from the column by isocratic elution (100 mM aqueous ammonium acetate, 20 min). The obtained data was analyzed using MassLynx software (Waters Corporation). The column used was an ACQUITY UPLC Protein BEH SEC Column, 200 Å, 1.7 μm, 4.6 mm×150 mm (Waters), and the measurement was carried out at a flow rate of 0.1 to 0.3 mL / min and a column temperature of 25°C.

[0144] [Production Example 1: Antibody-drug conjugate 1 (ADC1)]

[0145] Step 1: Conjugation of antibody and drug linker To a solution containing 0.15 mg of antibody 1 prepared in Example 1, a DMSO solution containing 10 mM SMCC-DM1 (MedChemExpress) was added at room temperature so that the amount was 10 equivalents per antibody molecule, and DMA (N,N-dimethylacetamide) was added to a final concentration of 10%. The mixture was stirred at room temperature for 24 hours using a rotator to bind the drug linker to the antibody, yielding antibody-drug conjugate 1 (ADC1).

[0146] Step 2: Purification and Characterization The solution containing antibody-drug conjugate 1 (ADC1) obtained in step 1 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-1. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 0.255 mg / mL, average number of drugs bound per antibody molecule (n): 2.5

[0147] [Production Example 2: Antibody-drug conjugate 2 (ADC2)]

[0148] Step 1: Conjugation of antibody and drug linker To a solution containing 0.29 mg of antibody 2 prepared in Example 1, a DMSO solution containing 10 mM SMCC-DM1 (MedChemExpress) was added at room temperature so that the amount was 10 equivalents per antibody molecule, and DMA was added to a final concentration of 10%. The mixture was stirred at room temperature for 24 hours using a rotator to bind the drug linker to the antibody, yielding antibody-drug conjugate 2 (ADC2).

[0149] Step 2: Purification and Characterization The solution containing antibody-drug conjugate 2 (ADC2) obtained in step 1 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-1. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 2.24 mg / mL, average number of drugs bound per antibody molecule (n): 3.5

[0150] [Production Example 3: Antibody-drug conjugate 3 (ADC3)]

[0151] Step 1: Conjugation of antibody and drug linker To a solution containing 0.18 mg of antibody 3 prepared in Example 1, a DMSO solution containing 10 mM SMCC-DM1 (MedChemExpress) was added at room temperature so that the amount was 10 equivalents per antibody molecule, and DMA was added to a final concentration of 10%. The mixture was stirred at room temperature for 24 hours using a rotator to bind the drug linker to the antibody, yielding antibody-drug conjugate 3 (ADC3).

[0152] Step 2: Purification and Characterization The solution containing antibody-drug conjugate 3 (ADC3) obtained in step 1 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-1. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 0.182 mg / mL, average number of drugs bound per antibody molecule (n): 3.5

[0153] [Production Example 4: Antibody-drug conjugate 4 (ADC4)]

[0154] Step 1: Conjugation of antibody and drug linker To a solution containing 1 mg of antibody 4 prepared in Example 1, a DMSO solution containing 10 mM SMCC-DM1 (MedChemExpress) was added at room temperature to give 9.6 equivalents per antibody molecule, and DMA was added to a final concentration of 10%. The mixture was stirred at room temperature for 24 hours using a rotator to bind the drug linker to the antibody, yielding antibody-drug conjugate 4 (ADC4).

[0155] Step 2: Purification and Characterization The solution containing antibody-drug conjugate 4 (ADC4) obtained in step 1 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 0.503 mg / mL, average number of drugs bound per antibody molecule (n): 2.9

[0156] [Production Example 5: Antibody-drug conjugate 5 (ADC5)]

[0157] Step 1: Conjugation of antibody and drug linker To a solution containing 1 mg of antibody 4 prepared in Example 1, a DMSO solution containing 10 mM SPDB-DM4 (MedChemExpress) was added at room temperature to give 16 equivalents per antibody molecule, and DMA was added to a final concentration of 20%. The mixture was stirred at room temperature for 24 hours using a rotator to bind the drug linker to the antibody, yielding antibody-drug conjugate 5 (ADC5).

[0158] Step 2: Purification and Characterization The solution containing antibody-drug conjugate 5 (ADC5) obtained in step 1 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 0.500 mg / mL, average number of drugs bound per antibody molecule (n): 3.1

[0159] [Production Example 6: Antibody-drug conjugate 6 (ADC6)]

[0160] Step 1: Reduction of antibody A 5 mM aqueous TCEP solution (20 equivalents per antibody molecule) was added to a solution containing 64.8 mg of antibody 4 prepared in Example 1, and the mixture was incubated at 37°C for 2 hours to reduce the disulfide bond in the hinge region of the antibody.

[0161] Step 2: Conjugation of antibody and drug linker To the solution obtained in step 1, a DMSO solution containing 10 mM MC-VC-PAB-MMAE (MedChemExpress) (10 equivalents per antibody molecule) was added at room temperature, and the mixture was stirred using a rotator at 4°C for 24 hours to bind the drug linker to the antibody, thereby obtaining antibody-drug conjugate 6 (ADC6).

[0162] Step 3: Purification and Characterization The solution containing antibody-drug conjugate 6 (ADC6) obtained in Step 2 was purified, concentrated, and its concentration measured by common procedures A and C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 4.38 mg / mL, average number of drugs bound per antibody molecule (n): 6.8

[0163] [Production Example 7: Antibody-drug conjugate 7 (ADC7)]

[0164] Step 1: Reduction of antibody A 5 mM aqueous TCEP solution (30 equivalents per antibody molecule) was added to a solution containing 6.5 mg of antibody 4 prepared in Example 1, and the mixture was incubated at 37°C for 2 hours to reduce the disulfide bond in the hinge region of the antibody.

[0165] Step 2: Conjugation of antibody and drug linker To 1 mg of antibody in the above solution, a DMSO solution containing 10 mM MC-GGFG-Dxd (MedChemExpress) (9.6 equivalents per antibody molecule) was added at room temperature, and the mixture was stirred using a rotator at 4°C for 24 hours to bind the drug linker to the antibody, thereby obtaining antibody-drug conjugate 7 (ADC7).

[0166] Step 3: Purification and Characterization The solution containing antibody-drug conjugate 7 (ADC7) obtained in Step 2 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 1.17 mg / mL, average number of drugs bound per antibody molecule (n): 6.5

[0167] [Production Example 8: Antibody-drug conjugate 8 (ADC8)]

[0168] Step 1: Reduction of antibody A 5 mM aqueous TCEP solution (30 equivalents per antibody molecule) was added to a solution containing 6.5 mg of antibody 4 prepared in Example 1, and the mixture was incubated at 37°C for 2 hours to reduce the disulfide bond in the hinge region of the antibody.

[0169] Step 2: Conjugation of antibody and drug linker To 1 mg of the antibody in the solution obtained in step 1, a DMSO solution containing 10 mM MC-GGFG-Dx8951 (MedChemExpress) (9.6 equivalents per antibody molecule) was added at room temperature, and the mixture was stirred using a rotator at 4°C for 24 hours to bind the drug linker to the antibody, thereby obtaining antibody-drug conjugate 8 (ADC8).

[0170] Step 3: Purification and Characterization The solution containing antibody-drug conjugate 8 (ADC8) obtained in Step 2 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 1.07 mg / mL, average number of drugs bound per antibody molecule (n): 4.9

[0171] [Production Example 9: Antibody-drug conjugate 9 (ADC9)]

[0172] Step 1: Reduction of antibody A 5 mM aqueous TCEP solution (30 equivalents per antibody molecule) was added to a solution containing 6.5 mg of antibody 4 prepared in Example 1, and the mixture was incubated at 37°C for 2 hours to reduce the disulfide bond in the hinge region of the antibody.

[0173] Step 2: Conjugation of antibody and drug linker To 1 mg of the antibody in the solution obtained in step 1, a DMSO solution containing 10 mM MC-VC-PAB-MMAF (MedChemExpress) (9.6 equivalents per antibody molecule) was added at room temperature, and the mixture was stirred using a rotator at 4°C for 24 hours to bind the drug linker to the antibody, thereby obtaining antibody-drug conjugate 9 (ADC9).

[0174] Step 3: Purification and Characterization The solution containing antibody-drug conjugate 9 (ADC9) obtained in Step 2 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 1.64 mg / mL, average number of drugs bound per antibody molecule (n): 1.2

[0175] [Production Example 10: Antibody-drug conjugate 10 (ADC10)]

[0176] Step 1: Reduction of antibody A 5 mM aqueous TCEP solution (30 equivalents per antibody molecule) was added to a solution containing 6.5 mg of antibody 4 prepared in Example 1, and the mixture was incubated at 37°C for 2 hours to reduce the disulfide bond in the hinge region of the antibody.

[0177] Step 2: Conjugation of antibody and drug linker To 1 mg of the antibody in the solution obtained in step 1, a DMSO solution containing 10 mM MC-CL2-SN38 (MedChemExpress) (9.6 equivalents per antibody molecule) was added at room temperature, and the mixture was stirred using a rotator at 4°C for 24 hours to bind the drug linker to the antibody, thereby obtaining antibody-drug conjugate 10 (ADC10).

[0178] Step 3: Purification and Characterization The solution containing antibody-drug conjugate 10 (ADC10) obtained in Step 2 was purified and its concentration measured by common procedure C, and the following characteristic values ​​were obtained by common procedure D-2. The product was confirmed to be free of aggregates by dynamic light scattering. Antibody concentration: 1.65 mg / mL, average number of drugs bound per antibody molecule (n): 4.0

[0179] Example 5: Evaluation of binding ability of ADCs to multiple cancer cell lines The binding ability and pH dependency of ADCs 1 to 10 prepared in Example 4 and antibodies 1 to 4 prepared in Example 1 to the gastric cancer cell line NUGC-4 were evaluated by flow cytometry in the same manner as in Example 2. The antibodies and ADCs were serially diluted starting from 300 μg / mL at a common ratio of 3, and the median fluorescence intensity (MFI, mean fluorescence intensity) at pH 7.4 and pH 6.0 was evaluated. The acquired data was analyzed using XLfit software (IDBS), and the EC ５０ As a result, no decrease in activity was observed due to ADC formation for any of Antibodies 1 to 4 (Figure 5).

[0180] Example 6: Ex vivo plasma stability evaluation of ADCs ADCs 6 to 10 prepared in Test Example 4 were evaluated for stability in mouse and human plasma. Each ADC, adjusted to 1 mg / mL, was added to human and BALB / c mouse plasma (anticoagulant: Heparin Na) to a final concentration of 20 μg / mL, and the mixture was incubated for 48 hours. The incubation samples were deproteinized with ethanol or acetonitrile / methanol (9 / 1, v / v), and the free drug from the extracted ADC was measured using LC / MS / MS. Free payload release (%) was calculated using the following formula: As a result, ADCs 6 to 9 were found to be stable in mouse and human plasma (FIG. 6).

[0181] Example 7: Cytotoxicity mediated by ADCs ADCs 1 to 3 were prepared using the procedure described in Example 4, and their cytotoxicity against the gastric cancer cell line NUGC-3 was evaluated using the CellTiter-Glo 2.0 luminescent cell viability assay (Promega). Each type of cell was seeded at 1,000 cells / well in a 96-well plate, and the ADCs were added and cultured at 37°C for 5 days. After 5 days, the CellTiter-Glo 2.0 luminescent cell viability assay was performed according to the manufacturer's recommendations. The relative luminescence intensity was measured using an EnSpire Multimode Plate Reader (PerkinElmer), and the data was analyzed using XLfit software (IDBS). The IC ５０ The results showed that although antibodies 1 to 3 alone did not exhibit cytotoxicity against NUGC-3, when antibodies 1 to 3 were converted into ADCs, all of the ADCs exhibited cytotoxicity against NUGC-3, suggesting the usefulness of the ADCs of the present invention as antitumor agents.

[0182] Furthermore, ADCs 4 to 10 with different drug linkers were prepared using the procedure described in Example 4, and their cytotoxicity in a pancreatic cancer cell line (Capan-1) and a gastric cancer cell line (NUGC-4) was evaluated using the same method as described above (Figure 8). As a result, it was confirmed that all Examples except ADC7 exhibited growth inhibitory effects against NUGC-4 and Capan-1. Note that the drug linker used in ADC7, MC-GGFG-Dxd, is the same as the drug linker used in, for example, trastuzumab deruxtecan (an anti-HER2 antibody ADC), which is marketed as an antitumor agent, and this does not mean that the drug linker itself is inactive. In other words, even if a drug linker has activity, it does not necessarily exhibit activity when converted into an ADC from the perspective of its relationship with the antibody. Furthermore, ADC8, which uses MC-GGFG-Dx8951, a drug linker structurally similar to that of ADC7, exhibits a growth inhibitory effect, so it is difficult to predict whether or not the ADC will actually exhibit activity when converted into an ADC.

[0183] Example 8: Antitumor effect of ADC (in vivo) Cancer-bearing animal models were prepared using gastric cancer (NUGC-4) and pancreatic cancer (BxPC-3) cell lines. First, each cancer cell line was prepared and subcutaneously inoculated into the right thoracic region of nude mice (BxPC-3: 5 × 10 ６ pieces / 100ul, NUGC-4:2×10 ６ (cells / 100 μl). The volume of the inoculated tumor was 100-300 mm ３ When tumor size reached approximately 100%, nude mice were randomly assigned to a control group (administered with PBS) and a test group, and the ADC6 was intravenously injected at a dose of 5 mg / kg, 10 mg / kg, or 20 mg / kg. As shown in Figure 9, weekly administration, biweekly administration, and single administration were adopted as the administration frequency. The tumor size was measured twice a week, and calculated using the following formula: tumor size (mm ３ ) = (long axis x short axis ２ ) / 2

[0184] As shown in Figure 9, tumor growth was confirmed to be suppressed in the ADC6 administration group compared to the control group from 7 days after the start of the test. These results demonstrate the remarkable inhibitory effect of the ADC of the present invention on the growth of pancreatic cancer and gastric cancer. Furthermore, while target expression was observed in normal cell lines (MSC, HUVEC) under pH 6 conditions for antibody 4, no weight loss or the like was observed even in the high-dose ADC of the present invention administration group. These results suggest that the ADC of the present invention selectively binds to tumor tissue due to its pH dependency, and suggest that the ADC of the present invention is highly safe and useful for therapeutic purposes.

[0185] SEQ ID NO: 1: Amino acid sequence of CDR1 in the heavy chains of antibody 1 and antibody 4 SEQ ID NO: 2: Amino acid sequence of CDR2 in the heavy chains of antibody 1 and antibody 4 SEQ ID NO: 3: Amino acid sequence of CDR3 in the heavy chains of antibody 1 and antibody 4 SEQ ID NO: 4: Amino acid sequence of CDR1 in the light chains of antibody 1 and antibody 4 LGS: Amino acid sequence of CDR2 in the light chains of antibody 1 and antibody 4 SEQ ID NO: 6: Amino acid sequence of CDR3 in the light chains of antibody 1 and antibody 4 SEQ ID NO: 7: Amino acid sequence of CDR1 in the heavy chain of antibody 2 SEQ ID NO: 8: Amino acid sequence of CDR2 in the heavy chain of antibody 2 SEQ ID NO: 9: Amino acid sequence of CDR3 in the heavy chain of antibody 2 SEQ ID NO: 10: Amino acid sequence of CDR1 in the light chain of antibody 2 AAS: Amino acid sequence of CDR2 in the light chain of antibody 2 SEQ ID NO: 12: Amino acid sequence of CDR3 in the light chain of antibody 2 SEQ ID NO: 13: Amino acid sequence of CDR1 in the heavy chain of antibody 3 SEQ ID NO: 14: Amino acid sequence of CDR2 of the heavy chain of antibody 3 SEQ ID NO: 15: Amino acid sequence of CDR3 of the heavy chain of antibody 3 SEQ ID NO: 16: Amino acid sequence of CDR1 of the light chain of antibody 3 WAS: Amino acid sequence of CDR2 of the light chain of antibody 3 SEQ ID NO: 18: Amino acid sequence of CDR3 of the light chain of antibody 3 SEQ ID NO: 19: Nucleotide sequence of cDNA encoding the heavy chain variable region of antibody 1 SEQ ID NO: 20: Amino acid sequence of the heavy chain variable region of antibody 1 SEQ ID NO: 21: Nucleotide sequence of cDNA encoding the light chain variable region of antibody 1 SEQ ID NO: 22: Amino acid sequence of the light chain variable region of antibody 1 SEQ ID NO: 23: Nucleotide sequence of cDNA encoding the heavy chain variable region of antibody 2 SEQ ID NO: 24: Amino acid sequence of the heavy chain variable region of antibody 2 SEQ ID NO: 25: Nucleotide sequence of cDNA encoding the light chain variable region of antibody 2 SEQ ID NO: 26: Amino acid sequence of the light chain variable region of antibody 2 SEQ ID NO: 27: Nucleotide sequence of cDNA encoding the heavy chain variable region of antibody 3 SEQ ID NO: 28: Amino acid sequence of the heavy chain variable region of antibody 3 SEQ ID NO: 29: Nucleotide sequence of cDNA encoding the light chain variable region of antibody 3 SEQ ID NO: 30: Amino acid sequence of the light chain variable region of antibody 3 SEQ ID NO: 31: Nucleotide sequence of cDNA encoding the heavy chain variable region of antibody 4 SEQ ID NO: 32: Amino acid sequence of the heavy chain variable region of antibody 4 SEQ ID NO: 33: Nucleotide sequence of cDNA encoding the light chain variable region of antibody 4 SEQ ID NO: 34: Amino acid sequence of the light chain variable region of antibody 4SEQ ID NO: 35: Nucleotide sequence of the heavy chain of antibody 1 SEQ ID NO: 36: Amino acid sequence of the heavy chain of antibody 1 SEQ ID NO: 37: Nucleotide sequence of the light chain of antibody 1 SEQ ID NO: 38: Amino acid sequence of the light chain of antibody 1 SEQ ID NO: 39: Nucleotide sequence of the heavy chain of antibody 2 SEQ ID NO: 40: Amino acid sequence of the heavy chain of antibody 2 SEQ ID NO: 41: Nucleotide sequence of the light chain of antibody 2 SEQ ID NO: 42: Amino acid sequence of the light chain of antibody 2 SEQ ID NO: 43: Nucleotide sequence of the heavy chain of antibody 3 SEQ ID NO: 44: Amino acid sequence of the heavy chain of antibody 3 SEQ ID NO: 45: Nucleotide sequence of the light chain of antibody 3 SEQ ID NO: 46: Amino acid sequence of the light chain of antibody 3 SEQ ID NO: 47: Nucleotide sequence of the heavy chain of antibody 4 SEQ ID NO: 48: Amino acid sequence of the heavy chain of antibody 4 SEQ ID NO: 49: Nucleotide sequence of the light chain of antibody 4 SEQ ID NO: 50: Amino acid sequence of the light chain of antibody 4 All publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. An antibody that specifically binds to sulfated glycosaminoglycans, selected from at least one of the following groups (1) to (3): (1) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, An antibody comprising a light chain containing CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 6; (2) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, An antibody comprising a light chain containing CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 10, CDR2 consisting of the amino acid sequence shown in AAS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 12; and (3) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, An antibody comprising a light chain containing CDR1, consisting of the amino acid sequence shown in SEQ ID NO: 16; CDR2, consisting of the amino acid sequence shown in WAS; and CDR3, consisting of the amino acid sequence shown in SEQ ID NO:

18.

2. The antibody according to claim 1, which is at least one selected from the group consisting of (1) to (4) below: (1) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 20 and having binding activity to sulfated glycosaminoglycans, and An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 22 and having binding activity to sulfated glycosaminoglycans; (2) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 24 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 26, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 26 and having binding activity to sulfated glycosaminoglycans; (3) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 28 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 30, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 30 and having binding activity to sulfated glycosaminoglycans; and (4) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and having binding activity to sulfated glycosaminoglycans.

3. The antibody according to claim 1, which is at least one selected from the group consisting of (1) to (4) below: (1) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 36 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 38 and possessing binding activity to sulfated glycosaminoglycans; (2) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 40 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 42 and possessing binding activity to sulfated glycosaminoglycans; (3) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 44 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 46 and having binding activity to sulfated glycosaminoglycans; and (4) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48, or a heavy chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 50 and possessing binding activity to sulfated glycosaminoglycans.

4. The antibody according to claim 1, which is a human antibody.

5. The antibody according to claim 1, wherein the subclass is an IgG1 antibody.

6. The antibody according to claim 1, characterized by binding with high affinity in the acidic pH range.

7. Antibody-drug conjugates (ADCs) represented by the following general formula 1: Ab-(LD)p (1) [In the formula, Ab is an antibody that specifically binds to sulfated glycosaminoglycans, which is selected from at least one of the following groups (1) to (3): (1) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, An antibody comprising a light chain containing CDR1, consisting of the amino acid sequence shown in SEQ ID NO: 4; CDR2, consisting of the amino acid sequence shown in LGS; and CDR3, consisting of the amino acid sequence shown in SEQ ID NO:

6. (2) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 7, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 9, An antibody comprising a light chain containing CDR1, consisting of the amino acid sequence shown in SEQ ID NO: 10; CDR2, consisting of the amino acid sequence shown in AAS; and CDR3, consisting of the amino acid sequence shown in SEQ ID NO:

12. (3) A heavy chain comprising CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 13, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 15, An antibody comprising a light chain containing CDR1, consisting of the amino acid sequence shown in SEQ ID NO: 16; CDR2, consisting of the amino acid sequence shown in WAS; and CDR3, consisting of the amino acid sequence shown in SEQ ID NO:

18. L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyrate-N-hydroxysuccinimidone (SPDB), or MC-CL2. D is monomethyl auristatin, maytansinoid, or camptothecin derivative. p is an integer selected from 1 to 12.

8. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is at least one selected from the group consisting of (1) to (4) below: (1) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 20 and having binding activity to sulfated glycosaminoglycans, and An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 22, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 22 and having binding activity to sulfated glycosaminoglycans; (2) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 24, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 24 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 26, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 26 and having binding activity to sulfated glycosaminoglycans; (3) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 28, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 28 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 30, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 30 and having binding activity to sulfated glycosaminoglycans; and (4) A heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, or a heavy chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 32 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 34, or a light chain containing a variable region having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 34 and having binding activity to sulfated glycosaminoglycans.

9. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is at least one selected from the group consisting of (1) to (4) below: (1) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 36 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 38, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 38 and possessing binding activity to sulfated glycosaminoglycans; (2) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 40, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 40 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 42, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 42 and possessing binding activity to sulfated glycosaminoglycans; (3) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 44, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 44 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 46, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 46 and having binding activity to sulfated glycosaminoglycans; and (4) A heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48, or a heavy chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 48 and having binding activity to sulfated glycosaminoglycans, An antibody comprising a light chain consisting of the amino acid sequence shown in SEQ ID NO: 50, or a light chain having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 50 and having binding activity to sulfated glycosaminoglycans.

10. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody comprising a heavy chain containing CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 1, CDR2 consisting of the amino acid sequence shown in SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO: 3, and a light chain containing CDR1 consisting of the amino acid sequence shown in SEQ ID NO: 4, CDR2 consisting of the amino acid sequence shown in LGS, and CDR3 consisting of the amino acid sequence shown in SEQ ID NO:

6.

11. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody comprising a heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO:

22.

12. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

38.

13. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody comprising a heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO:

34.

14. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

50.

15. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an IgG1 antibody.

16. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is a human antibody.

17. The antibody-drug conjugate (ADC) according to claim 7, wherein Ab is an antibody that binds with higher affinity in the acidic pH region than in the neutral pH region.

18. The antibody-drug conjugate (ADC) according to claim 7, wherein L is 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB), 6-maleimidocaproyl-glycine-glycine-phenylalanine-glycine (MC-GGFG), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), 4-(2-pyridyldithio)butyrate-N-hydroxysuccinimid ester (SPDB), or MC-CL2.

19. The antibody-drug conjugate (ADC) according to claim 7, wherein D is MMAE, MMAF, DM1, DM4, Dx8951, Dxd, or SN-38.

20. The antibody-drug conjugate (ADC) according to claim 7, wherein L-D is SMCC-DM1, SPDB-DM4, MC-Val-Cit-PAB-MMAE, MC-Val-Cit-PAB-MMAF, MC-GGFG-Dx8591, or MC-CL2-SN-38.

21. Ab is an antibody comprising a heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO:

22. The antibody-drug conjugate (ADC) according to claim 7, wherein L-D is MC-Val-Cit-PAB-MMAE.

22. Ab is an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

38. L-D is MC-Val-Cit-PAB-MMAE, and the antibody-drug conjugate (ADC) is as described in claim 7.

23. Ab is an antibody comprising a heavy chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO: 32, and a light chain containing a variable region consisting of the amino acid sequence shown in SEQ ID NO:

34. The antibody-drug conjugate (ADC) according to claim 7, wherein L-D is MC-Val-Cit-PAB-MMAE.

24. Ab is an antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 48 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

50. L-D is MC-Val-Cit-PAB-MMAE, and the antibody-drug conjugate (ADC) is as described in claim 7.

25. The antibody-drug conjugate (ADC) according to claim 7, wherein p is 3 to 8.

26. A pharmaceutical composition comprising an antibody-drug conjugate (ADC) according to any one of claims 7 to 25, and a carrier.

27. An antitumor agent containing an antibody-drug conjugate (ADC) according to any one of claims 7 to 25 as an active ingredient.

28. The antitumor agent according to claim 27 for treating cancer expressing sulfated glycosaminoglycans.

29. The antitumor agent according to claim 27 for treating gastric cancer, pancreatic cancer, breast cancer, colorectal cancer, or lung cancer.