Pharmaceutical composition for treating and / or preventing cancer

Abstract

A conjugate in which an antibody having immunoreactivity with a CAPRIN-1 protein having an amino acid sequence represented by any one of SEQ ID NO: 2 to SEQ ID NO: 30 or an amino acid sequence having 80% or more sequence identity with the amino acid sequence, or a fragment thereof, is bound to benzodiazepine # imgabs0 # is useful as an antibody-drug conjugate (ADC).

Description

Technical Field

[0001] The present invention relates to antibodies or fragments thereof against CAPRIN-1 protein and benzodiazepines The present invention relates to a conjugate of the same type, and a medical use of the conjugate as a therapeutic and / or preventive agent for cancer. Background Art

[0002] Various antibody drugs that target specific antigenic proteins on cancer cells are used in cancer treatment as cancer therapeutic drugs with few side effects due to their cancer specificity. For example, cytoplasmic-activation and proliferation-associated protein 1 (CAPRIN-1) is expressed on the cell membrane surface of many solid cancers, and antibodies against the CAPRIN-1 protein are known to be promising as drugs for the treatment and / or prevention of cancer (Patent Document 1).

[0003] In addition, benzodiazepines are known to The class of pyrrolobenzodiazepines is a substance that binds to the minor groove of DNA by recognizing a specific DNA sequence, thereby inhibiting DNA synthesis and inhibiting cell proliferation, thereby exerting anti-tumor activity. Specific examples include pyrrolobenzodiazepines. PBD, indolebenzodiazepine IGN, Pyridobenzodiazepine Class (PDD).

[0004] In recent years, research has been conducted to enhance the efficacy of antibody drugs against cancer, and in particular, the development of antibody-drug conjugates (ADCs) in which drugs that have a strong ability to directly kill cells are conjugated to antibodies has become popular (Non-Patent Documents 1 and 2). There are few successful examples of ADCs of this type, but ZYNLONTA, an ADC in which Tesirine (a derivative of SG3249), one of the PBD derivatives, is linked to an antibody targeting CD19, has been successfully developed. TM(loncastuximab tesirine-lpyl) has been approved for use in large B-cell lymphoma, and has achieved a complete or partial response of more than 48% for relapsed / refractory diffuse large B-cell lymphoma (DLBCL). In addition, Rovalpituzumab Tesirine, which is a DLL-3 antibody linked to Tesirine, Vadastuximab Talirine (SGN-CD33A), which is a CD33 antibody linked to Talirine, Camidanlumab Tesirine (Cami), which is a CD25 antibody linked to Tesirine, and SGN-CD70a, which is a CD70a antibody linked to SGD-1882.

[0005] Prior art literature

[0006] Patent Literature

[0007] Patent Document 1: WO2010 / 016526

[0008] Non-patent literature

[0009] Non-patent literature 1: Lancet Oncol. 2016; 17: e256-62

[0010] Non-patent literature 2: Pharm Res. 2015 Nov; 32(11): 3526-40 Summary of the invention

[0011] Problems to be solved by the invention

[0012] As mentioned above, benzodiazepines have been used to enhance the efficacy of antibody drugs against cancer. ADCs of this type have been put into practical use, but their success has been limited to certain cancers, and their anti-tumor effects are also limited.

[0013] Therefore, the present invention is based on the use of benzodiazepine The purpose of this class of ADCs is to enhance the anti-tumor effect compared with existing technologies.

[0014] Means for solving problems

[0015] As a result of intensive research, the inventors found that antibodies against CAPRIN-1 protein and benzodiazepines The conjugates of the present invention have a much stronger anti-tumor effect than the antibodies or fragments thereof directed against the CAPRIN-1 protein alone, and further, benzodiazepines are conjugated to the antibodies or fragments thereof directed against the CAPRIN-1 protein. The enhancement of the anti-tumor effect of the same class of drugs by conjugating benzodiazepines to existing cancer antibody drugs The present invention was completed because the enhancement effect of the anti-tumor effect is significantly superior to that of the same type.

[0016] Specifically, the present invention has the following features (1) to (15).

[0017] (1) Antibodies or fragments thereof and benzodiazepines The antibody or fragment thereof is immunoreactive with a CAPRIN-1 protein having an amino acid sequence represented by any of the even-numbered sequence numbers from SEQ ID NOs. 2 to 30, or an amino acid sequence having a sequence identity of 80% or more to the amino acid sequence.

[0018] (2) The conjugate according to (1), wherein the antibody or fragment thereof is immunoreactive with a partial polypeptide of a CAPRIN-1 protein, and the partial polypeptide of the CAPRIN-1 protein has an amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, and 309, or an amino acid sequence having a sequence identity of 80% or more thereto.

[0019] (3) The conjugate according to claim 1 or 2, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

[0020] (4) The conjugate according to any one of (1) to (3), wherein the antibody or fragment thereof is any one of the following (A) to (M),

[0021] (A) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 36, 37 and 38 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 40, 41 and 42 as CDR1, CDR2 and CDR3, respectively,

[0022] (B) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 44, 45 and 46 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 48, 49 and 50 as CDR1, CDR2 and CDR3, respectively,

[0023] (C) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 52, 53 and 54 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 56, 57 and 58 as CDR1, CDR2 and CDR3, respectively,

[0024] (D) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 60, 61 and 62 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 64, 65 and 66 as CDR1, CDR2 and CDR3, respectively,

[0025] (E) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 170, 171 and 172 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 173, 174 and 175 as CDR1, CDR2 and CDR3, respectively,

[0026] (F) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementarity determining regions of SEQ ID NOs: 176, 177 and 178 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 as CDR1, CDR2 and CDR3, respectively,

[0027] (G) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 182, 183 and 184 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 185, 186 and 187 as CDR1, CDR2 and CDR3, respectively,

[0028] (H) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 188, 189 and 190 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 191, 192 and 193 as CDR1, CDR2 and CDR3, respectively,

[0029] (I) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 146, 147 and 148 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 149, 150 and 151 as CDR1, CDR2 and CDR3, respectively,

[0030] (J) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 272, 273 and 274 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 275, 276 and 277 as CDR1, CDR2 and CDR3, respectively,

[0031] (K) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 290, 291 and 292 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 293, 294 and 295 as CDR1, CDR2 and CDR3, respectively,

[0032] (L) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 301, 302, and 303 as CDR1, CDR2, and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 305, 306, and 307 as CDR1, CDR2, and CDR3, respectively,

[0033] (M) An antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementarity determining regions of SEQ ID NOs: 134, 135 and 136 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of SEQ ID NOs: 137, 138 and 139 as CDR1, CDR2 and CDR3, respectively.

[0034] (5) The conjugate according to any one of (1) to (4), wherein the antibody or fragment thereof is any one of the following (a) to (a1),

[0035] (a) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43,

[0036] (b) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:47 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:51,

[0037] (c) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59,

[0038] (d) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 67,

[0039] (e) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69,

[0040] (f) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71,

[0041] (g) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73,

[0042] (h) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75,

[0043] (i) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77,

[0044] (j) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79,

[0045] (k) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81,

[0046] (l) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83,

[0047] (m) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85,

[0048] (n) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87,

[0049] (o) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89,

[0050] (p) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:91,

[0051] (q) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:93,

[0052] (r) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:95,

[0053] (s) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:97,

[0054] (t) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:99,

[0055] (u) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101,

[0056] (v) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103,

[0057] (w) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105,

[0058] (x) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107,

[0059] (y) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109,

[0060] (z) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111,

[0061] (aa) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113,

[0062] (ab) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115,

[0063] (ac) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117,

[0064] (ad) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119,

[0065] (ae) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121,

[0066] (af) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123,

[0067] (ag) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125,

[0068] (ah) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127,

[0069] (ai) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129,

[0070] (aj) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131,

[0071] (ak) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133,

[0072] (a1) An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 304.

[0073] (6) The conjugate according to any one of (1) to (5), wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single-chain antibody.

[0074] (7) The conjugate according to any one of (1) to (6), wherein the antibody or fragment thereof is combined with a benzodiazepine The classes are connected by a linker.

[0075] (8) The conjugate according to any one of (1) to (7), wherein the benzodiazepine Benzodiazepines with antitumor activity kind.

[0076] (9) The conjugate according to any one of (1) to (8), wherein the benzodiazepine Pyrrolobenzodiazepine PBD, indolebenzodiazepine IGN, Pyridobenzodiazepine PDD or isoquinoline benzodiazepine Class (IQB).

[0077] (10) The conjugate according to any one of (1) to (9), wherein the benzodiazepine The classes are DSB-120, SJG-136 (SG2000), DC-81, DSB-120, SJG-136, SG2057, SG2202, SG2285, SGD-1882, SGD-1910, SG3199, SG3249, SG2219, IMGN779, IMGN632, (S)-N-(4-aminophenyl)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazo -3-yl)oxy)butyrylamide)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide, D211, D221, D231, GWL-78, KMR-28-39, or derivatives thereof.

[0078] (11) The conjugate according to (10), wherein the derivative is Tesirine which is a derivative of SG3249, Talirine which is a derivative of SGD-1910, SG3364, SG3227, SG3140, i.e. MC-Phe-Lys-PAB-SG2057, SG3170, SG3203, i.e. MC-Phe-Lys-PAB-SG2057, SG3231, SG3400, SG3376, DGN642, DGN549, FGX5-67 or FGX-2-62, FGX11-38.

[0079] (12) A pharmaceutical composition for treating and / or preventing cancer, comprising the conjugate according to any one of (1) to (11) as an active ingredient.

[0080] (13) The pharmaceutical composition according to (12), wherein the cancer is a cancer that expresses CAPRIN-1 protein on the cell membrane surface.

[0081] (14) The pharmaceutical composition according to (12) or (13), wherein the cancer is breast cancer, kidney cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, bile duct cancer, sarcoma, mast cell tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, head and neck cancer or urothelial carcinoma.

[0082] (15) A method for treating and / or preventing cancer, comprising administering the conjugate according to any one of (1) to (11) or the pharmaceutical composition according to any one of (12) to (14) to a subject.

[0083] Effects of the Invention

[0084] The conjugate of the present invention not only exerts a much stronger anti-tumor effect than the antibody or fragment thereof directed against CAPRIN-1 protein alone, but also has a higher efficacy than the cancer antibody drugs and benzodiazepines known to date. In addition, by conjugating an antibody or a fragment thereof against the CAPRIN-1 protein with a benzodiazepine The enhanced anti-tumor effect of the drug is similar to that of conjugating existing cancer antibody drugs to benzodiazepines. The enhancement effect of the anti-tumor effect is also significant compared with the same time. Therefore, the conjugate involved in the present invention is effective for the treatment and prevention of cancer. DETAILED DESCRIPTION

[0085] The antibody or fragment thereof against CAPRIN-1 protein (hereinafter referred to as "anti-CAPRIN-1 antibody") used in the present invention and benzodiazepines The antitumor activity of the conjugates is evaluated by investigating the inhibition of tumor growth in cancer-bearing animals in vivo as described below.

[0086] The term "conjugate" in the present invention refers to an antibody and a benzodiazepine Antibodies and benzodiazepines The binding of classes can also be through a connector.

[0087] The anti-CAPRIN-1 antibody forming the above-mentioned conjugate involved in the present invention may be a monoclonal antibody or a polyclonal antibody, but preferably a monoclonal antibody. In addition, the conjugate of the present invention may be any type of antibody as long as it can exert anti-tumor activity, and may be a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, or a non-human animal antibody.

[0088] The benzodiazepines forming the above conjugates of the present invention The class is known as a substance that is toxic to cancer cells by recognizing a specific DNA sequence and binding to the minor groove of the DNA, thereby inhibiting DNA synthesis and suppressing cell proliferation.

[0089] In the present invention, the subject to be treated and / or prevented for cancer is a mammal such as a human, pet, livestock, or competitive animal, and a preferred subject is a human.

[0090] The following is about the anti-CAPRIN-1 antibody and benzodiazepine involved in the present invention. Class, anti-CAPRIN-1 antibodies and benzodiazepines The present invention describes a conjugate of the type, a pharmaceutical composition using the conjugate, and a method for treating and / or preventing cancer.

[0091] <Anti-CAPRIN-1 Antibody>

[0092] Among the CAPRIN-1 proteins having an amino acid sequence represented by any of the even-numbered sequence numbers among SEQ ID NOs: 2 to 30 and having immunoreactivity with the anti-CAPRIN-1 antibody used in the present invention, the amino acid sequences represented by SEQ ID NOs: 6, 8, 10, 12, and 14 are amino acid sequences of canine CAPRIN-1 proteins, the amino acid sequences represented by SEQ ID NOs: 2 and 4 are amino acid sequences of human CAPRIN-1 proteins, the amino acid sequence represented by SEQ ID NOs: 16 is an amino acid sequence of bovine CAPRIN-1 protein, the amino acid sequence represented by SEQ ID NOs: 18 is an amino acid sequence of horse CAPRIN-1 protein, the amino acid sequences represented by SEQ ID NOs: 20 to 28 are amino acid sequences of mouse CAPRIN-1 proteins, and the amino acid sequence represented by SEQ ID NOs: 30 is an amino acid sequence of chicken CAPRIN-1 protein.

[0093] In addition, the anti-CAPRIN-1 antibody used in the present invention may be an antibody that is immunoreactive with a variant of the CAPRIN-1 protein having a sequence identity of 80% or more, preferably 90% or more, more preferably 95% or more, and even more preferably 99% or more to the amino acid sequence represented by any of the even-numbered sequence numbers 2 to 30. Here, "% sequence identity" means the percentage (%) of identical amino acids (or bases) relative to the total number of amino acids (or bases) (including the number of gaps) when two sequences are aligned (arranged) so as to achieve the maximum similarity (or identity) with or without the introduction of gaps.

[0094] The anti-CAPRIN-1 antibody used to prepare the conjugate in the present invention refers to an antibody or an antigen-binding fragment thereof that is immunoreactive with the full length or fragments of CAPRIN-1 protein as an antigen. Here, "immunoreactivity" refers to the property of the antibody to bind to the CAPRIN-1 protein or a partial polypeptide thereof in vivo.

[0095] The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.

[0096] Polyclonal antibodies immunoreactive with the full length or fragments of CAPRIN-1 protein (anti-CAPRIN-1 polyclonal antibodies) can be obtained by, for example, immunizing mice, mice producing human antibodies, rats, rabbits, chickens, etc. with native CAPRIN-1 protein, or fusion proteins with GST or the like, or partial peptides thereof, and obtaining serum. The obtained serum can be obtained by, for example, passing through ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, affinity column coupled with CAPRIN-1 protein or partial peptide, etc.

[0097] The full length or fragments of the CAPRIN-1 protein used in the above immunization, the base sequence and amino acid sequence of CAPRIN-1 and its homologues can be obtained, for example, by registering in GenBank (NCBI, USA) and using algorithms such as BLAST and FASTA (Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90: 5873-5877, 1993; Altschul et al., Nucleic Acids Res. 25: 3389-3402, 1997). In addition, the method for producing the CAPRIN-1 protein can be obtained by referring to WO2014 / 012479, and cells expressing the CAPRIN-1 protein can also be used.

[0098] Monoclonal antibodies immunoreactive with the full length or fragments of CAPRIN-1 protein (anti-CAPRIN-1 monoclonal antibodies) can be obtained, for example, by administering breast cancer cells SK-BR-3 expressing CAPRIN-1, the full length or fragments of CAPRIN-1 protein, etc. to mice for immunization, and selecting clones producing anti-CAPRIN-1 monoclonal antibodies from the fusion cells (hybridomas) obtained by separating spleen cells from the mice and myeloma cells. Antibodies produced by the selected hybridomas can be obtained by the same method as the above-mentioned method for purifying polyclonal antibodies.

[0099] The antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies, and non-human animal antibodies.

[0100] Human antibodies can be obtained by sensitizing human lymphocytes infected with Epstein-Barr virus with proteins, protein-expressing cells or their lysates, fusing the sensitized lymphocytes with myeloma cells such as human-derived U266 cells, and obtaining antibodies immunoreactive with the full-length CAPRIN-1 protein or fragments thereof from the fused cells.

[0101] Humanized antibodies are altered antibodies also called reshaped human antibodies. Humanized antibodies are constructed by transplanting the complementary determining regions of antibodies derived from immune animals into the complementary determining regions of human antibodies. Gene recombination methods, which are general methods, are also well-known techniques. Specifically, by the PCR method, a DNA sequence designed to connect the complementary determining regions of, for example, mouse antibodies and rabbit antibodies to the framework regions of human antibodies is synthesized from several oligonucleotides made in a manner with overlapping portions at the ends. The obtained DNA is connected to the DNA encoding the constant region of the human antibody, then integrated into an expression vector, and introduced into the host to produce it, thereby obtaining a humanized antibody (refer to EP239400, WO96 / 02576). The framework region of the human antibody connected via the complementary determining region can select a framework region that allows the complementary determining region to form a good antigen binding site. If necessary, the amino acids in the framework region of the variable region of the antibody can also be replaced so that the complementary determining region of the reshaped human antibody forms an appropriate antigen binding site (Sato K. et al., Cancer Research 1993, 53: 851-856). Alternatively, the framework regions may be replaced with those derived from various human antibodies (see WO99 / 51743).

[0102] Antibodies are usually heteropolymeric glycoproteins containing at least 2 heavy chains and 2 light chains. Antibodies are composed of 2 identical light chains and 2 identical heavy chains. The heavy chain has a heavy chain variable region at one end, to which are attached several constant regions. The light chain has a light chain variable region at one end, to which are attached several constant regions. The variable region displays specific variable regions called complementarity determining regions (CDRs) that confer binding specificity to the antibody. The relatively conserved portions of the variable region are called framework regions (FRs). The variable regions of complete heavy and light chains contain 4 FRs connected by 3 CDRs (CDR1 to CDR3) respectively.

[0103] In addition, the sequences of the constant regions and variable regions of the heavy and light chains of human origin can be obtained from NCBI (United States: GenBank, UniGene, etc.). For example, the heavy chain constant region of human IgG1 can refer to the sequence of accession number J00228, the heavy chain constant region of human IgG2 can refer to the sequence of accession number J00230, the human light chain κ constant region can refer to the sequences of accession numbers V00557, X64135, X64133, etc., and the human light chain λ constant region can refer to the sequences of accession numbers X64132, X64134, etc.

[0104] Chimeric antibodies are antibodies produced by combining sequences derived from different animals, for example, antibodies containing the heavy chain variable region and light chain variable region of a mouse antibody and the constant region of the heavy chain variable region and light chain variable region of a human antibody, etc. The production of chimeric antibodies can be carried out using known methods, for example, by connecting DNA encoding the antibody V region with DNA encoding the human antibody C region, integrating it into an expression vector and introducing it into a host, thereby producing it.

[0105] Non-human animal antibodies can be obtained by immunizing animals with sensitizing antigens according to known methods. As a general method, sensitizing antigens can be injected into the peritoneal cavity, intradermal or subcutaneous of animals such as mice. When injecting sensitizing antigens, they can be mixed with various adjuvants represented by CFA (complete Freund's adjuvant) in appropriate amounts and administered to animals multiple times. After immunizing animals and confirming that the serum contains anti-CAPRIN-1 antibodies, serum can be obtained. As mentioned above, it can be activated by ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, affinity column coupled to CAPRIN-1 protein or partial peptide, etc. In addition, when obtaining monoclonal antibodies from non-human animals, immune cells can be collected from immunized animals and obtained by cell fusion with myeloma cells. The cell fusion of the immune cells and myeloma cells can be carried out according to known methods (see Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).

[0106] The antibodies used in the present invention can also be obtained as recombinant antibodies by cloning antibody genes from hybridomas, integrating them into appropriate vectors, introducing them into hosts, and producing them using gene recombination technology (see Carl, AK, Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990).

[0107] The anti-CAPRIN-1 antibody used to obtain the conjugate of the present invention may also be an antibody in which amino acids in the variable region (e.g., FR) or the constant region are replaced with other amino acids. The amino acid replacement is 1 or more, for example, less than 15, less than 10, 8 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less amino acids, preferably 1 to 9 amino acids. The replaced antibody should have the same or higher binding affinity to the antigen as the unsubstituted antibody, and should not cause rejection when used in humans.

[0108] The higher the binding affinity of the anti-CAPRIN-1 antibody used in the present invention to the CAPRIN-1 protein on the surface of cancer cells, the stronger the anti-tumor effect can be expected. The binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5×10 8 M -1 , at least 10 9 M -1 , at least 5×10 9 M -1 , at least 10 10 M -1 , at least 5×10 10 M -1 , at least 10 11 M -1 , at least 5×10 11 M -1 , at least 10 12 M -1 , or at least 10 13 M -1 .

[0109] The anti-CAPRIN-1 antibody used in the present invention can enhance the binding ability of the anti-CAPRIN-1 antibody to effector cells by replacing one, two or more amino acids in the heavy chain constant region of the antibody, or removing fucose bound to N-acetylglucosamine in the N-glycosidic bond sugar chain bound to the heavy chain constant region. The above may be a single amino acid replacement or a combination with an antibody bound to fucose.

[0110] Antibodies in which one, two or more amino acids in the heavy chain constant region are substituted can be prepared with reference to, for example, WO2004 / 063351, WO2011 / 120135, US Pat. No. 8,388,955, WO2011 / 005481, US Pat. No. 6,737,056, and WO2005 / 063351.

[0111] Antibodies from which fucose attached to N-acetylglucosamine in the N-glycosidic sugar chain in the heavy chain constant region has been removed, or cells producing the antibodies, can be prepared with reference to U.S. Patent No. 6,602,684, European Patent No. 1,914,244, and U.S. Patent No. 7,579,170. Antibodies from which fucose bound to N-acetylglucosamine in the N-glycosidic sugar chain bound to the heavy chain constant region has been removed, and compositions of antibodies to which fucose has been bound, or cells producing the compositions, can be prepared with reference to, for example, U.S. Patent No. 8,642,292.

[0112] The anti-CAPRIN-1 polyclonal antibody, anti-CAPRIN-1 monoclonal antibody, antibody preparation method, purification method, and preparation method of CAPRIN-1 protein or its partial polypeptide for immunization used in the present invention can refer to WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO 2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212.

[0113] Specific examples of the anti-CAPRIN-1 antibody in the present invention include the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212, but the following antibodies are mentioned as preferred anti-CAPRIN-1 antibodies.

[0114] An antibody or a fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as shown in SEQ ID NO: 2 or SEQ ID NO: 4, or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and even more preferably 99% or more) to the amino acid sequence.

[0115] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 31 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 36, 37, and 38, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 40, 41, and 42, or a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 140, 141, and 142, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 143, 144, and 145. 45 (CDR1, CDR2 and CDR3, respectively), and an antibody or fragment thereof that is immunoreactive with the CAPRIN-1 protein; or an antibody or fragment thereof that contains a heavy chain variable region comprising the complementary determining regions of SEQ ID NOs: 164, 165 and 166 (CDR1, CDR2 and CDR3, respectively), and a light chain variable region comprising the complementary determining regions of SEQ ID NOs: 167, 168 and 169 (CDR1, CDR2 and CDR3, respectively), and is immunoreactive with the CAPRIN-1 protein. More preferably, an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 39 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 43; or an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 70 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 71; or an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 78 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 79.

[0116] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 33 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 60, 61, and 62, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 64, 65, and 66, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 63 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 67 in the light chain variable region.

[0117] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 32 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 52, 53, and 54, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 56, 57, and 58, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 55 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 59 in the light chain variable region.

[0118] An antibody or a fragment thereof immunoreactive with a partial polypeptide of the CAPRIN-1 protein having the amino acid sequence shown in SEQ ID NO: 34 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 170, 171 and 172 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 173, 174 and 175 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein; or a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 176, 177 and 178 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein. More preferably, it is an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 80 and the light chain variable region comprises the amino acid sequence of sequence number 81; or an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 82 and the light chain variable region comprises the amino acid sequence of sequence number 83.

[0119] An antibody or fragment thereof immunoreactive with a partial polypeptide of the CAPRIN-1 protein having the amino acid sequence of SEQ ID NO: 35 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 182, 183 and 184 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 185, 186 and 187 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein; or a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 188, 189 and 190 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein. More preferably, it is an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 84 and the light chain variable region comprises the amino acid sequence of sequence number 85; or an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 86 and the light chain variable region comprises the amino acid sequence of sequence number 87.

[0120] An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 44, 45 and 46, and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 48, 49 and 50, and having immunoreactivity with the CAPRIN-1 protein. Preferably, the antibody or fragment thereof is one in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 47, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 51.

[0121] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 296 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 146, 147, and 148, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 149, 150, and 151, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 72 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 73 in the light chain variable region.

[0122] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 297 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 272, 273, and 274, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 275, 276, and 277. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 114 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 115 in the light chain variable region.

[0123] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 298 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 290, 291, and 292, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 293, 294, and 295, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 120 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 121 in the light chain variable region.

[0124] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 299 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 301, 302, and 303, and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 305, 306, and 307. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 300 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 304 in the light chain variable region.

[0125] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 308 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 134, 135, and 136, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 137, 138, and 139. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 68 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 69 in the light chain variable region.

[0126] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 309 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 134, 135, and 136, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 137, 138, and 139. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 68 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 69 in the light chain variable region.

[0127] In addition, the following anti-CAPRIN-1 antibodies are also preferably used.

[0128] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69.

[0129] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71.

[0130] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73.

[0131] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75.

[0132] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77.

[0133] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79.

[0134] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81.

[0135] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83.

[0136] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85.

[0137] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87.

[0138] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89.

[0139] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 91.

[0140] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 93.

[0141] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 95.

[0142] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 97.

[0143] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 99.

[0144] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101.

[0145] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103.

[0146] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105.

[0147] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107.

[0148] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109.

[0149] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111.

[0150] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113.

[0151] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115.

[0152] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117.

[0153] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119.

[0154] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121.

[0155] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123.

[0156] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125.

[0157] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127.

[0158] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129.

[0159] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131.

[0160] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133.

[0161] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 304.

[0162] In the Examples described below, conjugates with microtubule regulators were prepared using the above-mentioned polyclonal antibodies and monoclonal antibodies against the full-length CAPRIN-1 protein and a partial polypeptide region expressed on the cell membrane surface of cancer cells, and their strong antitumor effects were confirmed.

[0163] <Benzodiazepine Category >

[0164] Benzodiazepines There are substances in the class that can recognize specific DNA sequences and bind to the minor groove (DNA minor groove), inhibit DNA synthesis and inhibit cell proliferation to exert anti-tumor effects. In the present invention, benzodiazepines having such anti-tumor activity are preferably used. kind.

[0165] Benzodiazepines with antitumor activity Specific examples of the pyrrolobenzodiazepines include PBD, indolebenzodiazepine IGN, Pyridobenzodiazepine PDD or isoquinoline benzodiazepine Class (IQB).

[0166] PBD has an aromatic ring A (A ring), a ring B (B ring), and a pyrrolo ring C (C ring), and forms any of imine (N=C), methanolamine (NH-CH(OH)) or methanolamine methyl ether (NH-CH(OMe)) at the position of N10-N11 in ring B. In addition, PBD can also take the structure of C2-unsaturated PBD dimer, PBD dimer with C2 aromatic substitution, and polypyrrole-PBD. PBD has the ability to recognize and bind to a specific sequence of DNA, and the preferred sequence is 5'PuGPu3'.

[0167] Specific examples of PBD include anthramycin and its analogs, which are antitumor antibiotics. Anthramycin analogs include abbeymycin, chicamycin, mazethramycin, neothramycin A, meothramycin B, porothramycin, prothracarcin, sibanomicin (DC-102), sibiromycin, tomamycin, didehydroanhydroanthramycin, spadicomycin, and DC-81.

[0168] In addition, specific examples of PBD include compounds described in "Mantaj, J et al. 2016 Angew. Chem. Int Ed. 55, 2-29; D. Antonow and D. Thurston, Chem. Rev. 2011", "Leanna et al. J. Med. Chem. 2020, 63, 9603-9622", "Jhon et al. Expert Opinion on Biological Therapy Vol 21, 2021, 931-943", and "Julia et al. Angew Chem Int Ed Engl. 2017 Jan 9; 56 (2): 462-488".

[0169] Specific examples of the complex compound of PBD and linker include WO1993 / 018045, WO2000 / 046228, WO2002 / 083682, WO2005 / 040170, WO2005 / 110423, WO2005 / 085251, WO2006 / 135687, WO2010 / 043880, WO2013 / 055990, WO2013 / 055993, WO2014 / 057072, WO2014 / 159981, WO2014 / 057120, WO2015 / 052532, WO2015 / 181559, WO2017 / 020972, WO2 017 / 059289、WO2017 / 137555、WO2017 / 137556、WO2017 / 186894、WO2018 / 03 1662、WO2018 / 069490、WO2018 / 091646、WO2018 / 146188、WO2018 / 146189、WO 2018 / 18234、WO2018 / 192944、WO2019 / 034764、WO2019 / 065964、WO2019 / 09 6788、WO2019 / 104289、WO2019 / 126691、WO2019 / 224340、WO2020 / 006722、WO 2020 / 079229、WO2020 / 079239、WO2020 / 100954、WO2020 / 141923、WO2020 / 1 52462、WO2020 / 1964474、WO2020 / 196712、WO2021 / 137646、WO2022 / 063853、 WO2016 / 083468, WO2014 / 057073, WO2014 / 057113, WO2014 / 057114, WO2014 / 057115、WO2014 / 057117、WO2014 / 057118、WO2014 / 057119、WO2014 / 057120 、WO2014 / 057122、U.S. Patent No. 4316900、WO2020 / 245283、WO2019 / 197602、WO2004 / 043963、WO2005 / 085260、WO2006 / 111759、WO2010 / 010347、WO2010 / 043877、WO2011 / 128650、WO2011 / 130598、WO2011 / 130613、WO2011 / 130616、WO2013 / 041606、WO2013 / 053871、WO2013 / 053873、WO2013 / 055987、WO2013 / 053872、Compounds described in WO2013 / 164593, WO2013 / 164592, WO2014 / 096368, WO2014 / 140862, WO2014 / 140174, WO2015 / 052322, WO2015 / 052532, WO2015 / 052533, WO2015 / 052534, WO2015 / 052535, WO2016 / 038383, WO2016 / 037644, WO2016 / 044560, WO2017 / 137553, WO2017 / 129652, WO2017 / 223275, WO2018 / 146199, and WO2018 / 182341.

[0170] Examples of IGN and composite compounds of IGN and linkers include compounds described in WO2017 / 015495, WO2017 / 015496, WO2012 / 128868, WO2012 / 112687, WO2012 / 112708, WO2016 / 036801, WO2017 / 015502, WO2012 / 112708, WO2018 / 140435, WO2018 / 195245, WO2018 / 098258, WO2010 / 091150, WO2016 / 036804, WO2019 / 133652, WO2020102051, WO2020 / 102053, and WO2020 / 205564.

[0171] Examples of PDD and complex compounds of PDD and a linker include compounds described in WO2012 / 152915, WO2015 / 166289, WO2017 / 032983, WO2016 / 198869, WO2017 / 194960, WO2019 / 043417, WO2020 / 049286, WO2020 / 157491, and WO2022 / 023735.

[0172] Examples of IQB and complex compounds of IQB and a linker include compounds described in WO2016 / 149546, WO2018 / 071455, WO2018 / 053552, WO2017 / 011803, WO2017 / 091615, U.S. Patent No. 9974864, U.S. Patent No. 9504694, and U.S. Patent No. 10350218.

[0173] The benzodiazepines used in the present invention The class may be a monomer or a dimer. In the case of a dimer, in addition to a homodimer, it may also be a heterodimer with other antitumor compounds. The dimer may also be a heterodimer with benzodiazepines. A dimer in which the carbon atoms at the 8th, 7th or 2nd positions of the same class are bound to each other.

[0174] Benzodiazepines preferably used in the present invention DSB-120, SJG-136 (SG2000), DC-81, DSB-120, SJG-136, SG2057, SG2202, SG2285, SGD-1882, SGD-1910, SG3199, SG3249, SG2219, IMGN779, IMGN632, (S)-N-(4-aminophenyl)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazo -3-yl)oxy)butyrylamide)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide, D211, D221, D231, GWL-78, KMR-28-39 or a derivative thereof.

[0175] In addition, the derivative is preferably a derivative obtained by binding a linker to the compound. Specific examples include Tesirine (a derivative of SG3249), Talirine (a derivative of SGD-1910), SG3364, SG3227, SG3140 (MC-Phe-Lys-PAB-SG2057), SG3170, SG3203 (MC-Phe-Lys-PAB-SG2057), SG3231, SG3400, SG3376, DGN642, DGN549, FGX5-67, FGX-2-62, and FGX11-38.

[0176] < Anti-CAPRIN-1 Antibodies and Benzodiazepines Conjugates of the type >

[0177] In the present invention, anti-CAPRIN-1 antibody and benzodiazepine Conjugates of anti-CAPRIN-1 antibodies with benzodiazepines The binding form of the benzodiazepines is not particularly limited as long as it can maintain the anti-tumor activity against cancer, but it is preferably a binding form between the anti-CAPRIN-1 antibody and the benzodiazepines. The combination form of connecting structure between classes.

[0178] Here, the linker refers to a substance that can bind the anti-CAPRIN-1 antibody to the benzodiazepine Various known linkers can be used, or the structure on the activator side can be appropriately chemically modified to allow direct binding.

[0179] The type of linker and the details of the binding method can be determined according to known methods (for example, see Greg T. Hermanson Bioconjugate Techniques, Third Edition, WO2004 / 010957, and WO2014 / 012479).

[0180] In an embodiment of the present invention, anti-CAPRIN-1 antibody, benzodiazepine The following are examples of reactive groups attached to the class and the linker.

[0181] As the reactive groups attached to the amino acid sequence of the antibody or the glycoprotein of the modified amino acid, primary amine (ε-amino), carboxyl, thiol (sulfhydryl), carbonyl (ketone or aldehyde) and hydroxyl can be listed as long as no special chemical modification is performed. Primary amines are present at the N-terminus of polypeptides and the side chains of lysine residues. They are positively charged under physiological conditions and usually exist on the outside of proteins, so they can be used for binding without changing the structure of the protein. Carboxyl groups are present at the C-terminus of polypeptides and the side chains of aspartic acid and glutamic acid. Sulfhydryl groups are present on the side chains of cysteine ​​and form disulfide bonds that maintain the higher-order structure of proteins. Ketones or aldehydes are produced in glycoproteins by oxidizing sugar groups with sodium metaperiodate.

[0182] The conjugate of the present invention is obtained by combining a linker that is bound to the reactive group on the antibody with a benzodiazepine. Benzodiazepine Benzodiazepines or directly bind benzodiazepines to antibodies Class to make.

[0183] Specific examples of the reactive groups attached to PBD and the linker include the following groups.

[0184] As reactive groups capable of reacting with amines, N-hydroxysuccinimide (NHS) esters, imide esters, pentafluorophenyl esters, hydroxymethylphosphines, isothiocyanates, isocyanates, acyl azides, N-hydroxy esters, sulfonyl chlorides, aldehydes, glyoxal, epoxides, ethylene oxides, carbonates, aryls, carbodiimides, and carboxylic anhydrides.

[0185] Carbodiimide, diazoalkane, diazoacetyl compound, and carbonyldiimidazole are reactive groups capable of reacting with carboxyl groups and amines.

[0186] As the reactive group capable of reacting with a thiol group, there may be mentioned maleimide, haloacetamide, pyridinium disulfide, thiosulfone, vinyl sulfone, haloacetyl, aziridine, acryloyl, and aryl.

[0187] As the reactive group capable of reacting with aldehyde, hydrazide and alkoxyamine. As the reactive group capable of reacting with hydroxyl group, epoxy group, ethylene oxide, carbonyldiimidazole, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, isocyanate.

[0188] Isocyanate is a reactive group capable of reacting with a hydroxyl group.

[0189] As photoreactive groups, diazirine, aryl azide, aryl, benzophenol, and diazo compounds.

[0190] Specific examples of the linker having the above-mentioned reactive group include the following linkers.

[0191] As linkers having the same reactive group terminal, linkers having N-hydroxysuccinimide ester as a reactive group (e.g., disuccinimidyl glutarate (DSG), disuccinimidyl suberate (DSS), bis(sulfosuccinimidyl) suberate (BS3), tris(succinimidyl)aminotriacetate (TSAT), PEGylated bis(sulfosuccinimidyl) suberate (BS(PEG)5, BS(PEG)9), dithiobis(succinimidyl propionate) (DSP), 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP), ethylene glycol bis(succinimidyl succinate) (EGS), sulfo-ethylene glycol bis(succinimidyl succinate (sulfo-EGS), dimethyl adipimidate dihydrochloride (DMA), dimethyl pimelimidate dihydrochloride (D MP), dimethyl succinimidyl ester dihydrochloride (DMS), dimethyl 3,3'-dithiobispropanimidate dihydrochloride (DTBP), 1,5-difluoro-1,5-dinitrobenzene (DFDNB), disuccinimidyl tartrate (DST), bis[2-(succinimidyloxycarbonyloxy)ethyl]sulfone (BSOCOES), and linkers with maleimide as a reactive group (e.g., bismaleimidoethane (BMOE), 1,4-bismaleimidobutane (BMB), bismaleimidohexane (BMH), tris(2-maleimidoethyl)amine (TMEA), 1,8-bismaleimido-(PEG)2 (BM(PEG)2), 1,8-bismaleimido-(PEG)3 (BM(PEG)3), dithiobismaleimidoethane (DTME).

[0192] As the main linkers with different reactive group ends, linkers with NHS ester and maleimide as reactive groups (e.g., AMAS, BMPS, GMBS, Sulfo-MBS, MBS, Sulfo-MBS, SMCC, Sulfo-SMCC, EMCS, Sulfo-EMCS, SMPB, Sulfo-SMPB, SMPH, LC-SMCC, Sulfo-KMUS, SM(PEG)2, SM(PEG)4, SM(PEG)6, SM(PEG)8, SM(PEG)9) can be used. 12 、SM(PEG) 24 ), linkers with NHS esters and pyridyl dithiol as reactive groups (e.g., SPDP, LC-SPDP, sulfo-LC-SPDP, SMPT, (PEG)4SPDP, PEG12-SPDP), linkers with NHS esters and haloacetyl as reactive groups (e.g., SIA, SBAP, SIAB, sulfo-SIAB), linkers with NHS esters and aryl azide as reactive groups (e.g., ANB-NOS, sulfo-SANPAH, ATFB), linkers with NHS esters and diazirine as reactive groups (e.g., For example, SDA, sulfo-SDA, LC-SDA, SDAD, sulfo-SDAD), a linker with carbodiimide as a reactive group (e.g., DCC, EDC, EDAC, NHS, sulfo-NHS), a linker with maleimide and hydrazide as reactive groups (e.g., BMPH, EMCH, MPBH, KMUH), a linker with pyridyl dithiol and hydrazide as reactive groups (e.g., PDPH), a linker with isocyanate and maleimide as reactive groups (e.g., PMPI), and a linker with NHS ester and psoralen as reactive groups (e.g., SPB).

[0193] As other linkers, linkers comprising polypeptides can be used, such as Fmoc(fluorenylmethoxycarbonyl)-Ala-Ala-Asn-PAB, Fmoc-Ala-Ala-Asn(Trt)-PAB, Fmoc-PEG3-Ala-Ala-Asn(Trt)-PAB, Fmoc-PEG4-Ala-Ala-Asn(Trt)-PAB, Fmoc-Ala-Ala-Asn-PAB-PNP , Fmoc-Ala-Ala-Asn(Trt)-PAB-PNP, Fmoc-PEG3-Ala-Ala-Asn(Trt)-PAB-PNP, Azide(azide)-PEG4-Ala-Ala-Asn(Trt)-PAB-PNP, Mal(maleimide)-PE G4-Ala-Ala-Asn(Trt)-PAB-PNP, Fmoc-Val-Cit-PAB-OH, Val-Cit-PAB-OH, Fmoc-Val-Cit-PAB-PNP, MC-Val-Cit-PAB, MC-Val-Cit-PAB-PNP, etc.

[0194] In addition, Bis-PEG-acid (biscarboxyl polyethylene glycol), PEG Acid (carboxyl polyethylene glycol) (for example, Acid (carboxyl)-PEG-TEMPO, Amino (amino)-PEG-acid (carboxyl), Amino-PEG-CH2CO2H, Aminoxy (aminooxy)-PEG-acid, Azido (azido)-PEG-acid, Carboxy (carboxyl)-PEG-sulfonic acid (sulfonic acid), Fmoc (fluorenylmethyloxycarbonyl)-N-amido (amido)-PEG-acid, Fmoc-N-amido (amino)-PEG-CH2CO2H, Fmoc-aminooxy (aminooxy)-PEG-acid, Hydroxy (hydroxy)-PEG-acid, Hydroxy-PEG-CH2CO2H, m (methyl)-PEG-acid, m-PEG-(CH2)3-acid, Methoxytrityl (methoxytrityl)-N-PEG G-acid, N-Methyl(methyl)-N-(t-Boc(tert-butyloxycarbonyl))-PEG-acid, Propargyl(propargyl)-PEG-acid, Propargyl-PEG-CH2CO2H, Propargyl-PEG-(CH2)3-acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG-CH2CO2H, t-Boc-Aminooxy-PEG-acid, Acid-PEG-PFP ester (ester), Miscellaneous (mixed) PEG acid), PEG PFP ester (e.g., Acid-PEG-PFP ester, Bis-PEG-PFP ester), Bis-PEG-NHS, PEG Aldehyde (PEG aldehyde) (e.g., m-PEG-aldehyde (aldehyde), m-PEG-benzaldehyde (benzaldehyde), Ald (aldehyde group)-PEG-acid, Ald-PEG-amine, Ald-PEG-azide (azide), Ald-PEG-NH-Boc (butyloxycarbonyl), Ald-PEG-NHS ester, Ald-PEG-TFP ester, Ald-PEG-t-butyl (tert-butyl) ester), PEG Tosylate (PEG tosylate) (e.g.,Azido-PEG-Tos(Tosylate), Hydroxy-PEG-Tos, m-PEG-Tos, t-Boc-Aminooxy-PEG-Tos, Trifluoroethyl(trifluoroethyl)-PEG-Tos, Tos-PEG-acid, Tos-PEG-CH2CO2H, Tos-PEG-alkyne, Tos-PEG-t-butyl ester, Tos-PEG-CH2CO2tBu (tert-butyl), Tos-PEG-Tos, S-acetyl (acetyl) -PEG6-Tos, N-Tos-N- (t-butoxycarbonyl (tert-butyloxycarbonyl)) -animooxy-PEG4-Tos, Ms-PEG-Ms, Ms-PEG-t-butyl (tert-butyl) ester, PEG-Ms, Propargyl-PEG-Ms), Boc-PEG (e.g., Amino-PEG-t-Boc-Hydrazide (hydrazide), Azido-PEG-t-Boc-Hydrazide, Boc-NH-PEG-NH-Boc, Bromoac etamido(bromoacetamido)-PEG-Boc-amine(amine), m-PEG-ONHBoc, Mal(maleimide)-Alkyl(alkyl)-t-Boc-amine, N-Boc-PEG-alcohol(alcohol hydroxyl), N-Boc-PEG-bromide(bromine), N-methyl-N-(t-Boc)-PEG-acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG-CH2CO2H, t-Boc-N-Amido-PEG-amine, t-Boc-N-amido-PEG-azide, t-Boc-N-amido-PEG-NHS ester, t-Boc-N-amido-PEG-sulfonic acid), PEGNHS ester (e.g., Acid-PEG-NHS ester, Azido-PEG-NHS ester, Bis-PEG-NHS, Fmoc-PEG-NHS ester, m-PEG-NHS ester, m-PEG-NHS Carbonate, Mal-PEG-NHS ester, Propargyl-PEG-NHS ester, t-Boc-N-amido-PEG-NHS ester, t-Butoxycarbonyl (tert-butyloxycarbonyl)-PEG-NHS ester), Fmoc-PEG (e.g.,Fmoc-N-amido-PEG-acid, Fmoc-NH-PEG-CH2CO2H, Fmoc-PEG-NHS ester), Biotin (e.g., Biotin PEG-acid, Biotin PEG-alcohol, Biotin PEG-alkyne, Biotin PEG-amine, Biotin PEG-azide, Biotin PEG-DBCO, Biotin PEG-hydrazide, Biotin-PEG-Mal, Biotin-PEG-NHS, Biotin-EDA-PEG-NHS, Biotin-PEG-oxyamine, Biotin-PEG-PFP, Biotin-EDA-PEG-PFP, Biotin-PEG-Tetrazine, Biotin-PEG-TFP, Azide-SS-biotin, Biotin-PEG3-SS-azide, DBCO-S-S-PEG3-Biotin, Dde Biotin-PEG4-Alkyne, Dde Biotin-PEG4-Azide, Dde Biotin-PEG4-DBCO, Diazo Biotin-PEG3-Alkyne, Diazo Biotin-PEG3-Azide, Diazo Biotin-PEG3-DBCO, Diol Biotin-PEG3-Alkyne, Diol Biotin-PEG3-Azide, PC Biotin-PEG3-Alkyne, PC-Biotin-PEG4-PEG4-Alkyne, PC-Biotin-PEG4-PEG4-Alkyne, PC Biotin-PEG3-Azide, PC-Biotin-PEG4-PEG3-Azide, PC-Biotin-PEG4-NHS carbonate, PC DBCO-PEG3-Biotin, WSPCBiotin-PEG3-DBCO, Fmoc-Lys(biotin-PEG)-OH, Fmoc-N-amido-(PEG-biotin)-acid, TAMRA-Azide-PEG-Biotin), PEG Phosphonate, Aminooxy PEG (e.g.,Aminooxy-PEG-acid, Aminooxy-PEG-alcohol, Aminooxy-PEG-azide, Aminooxy-PEG-bromide, Aminooxy-PEG-methane, Aminooxy-PEG-Propargyl, Aminooxy-PEG-t-butyl ester, Aminooxy-PEG-Thiol, Bis-(Aminooxy)-PEG, t-Boc-Aminooxy-PEG-acid, t-Boc-Aminooxy-PEG-alcohol, t-Boc-Aminooxy-PEG-amine, t-Boc-Aminooxy-PEG-Azide, t-Boc-Aminooxy-PEG-Bromide, t-Boc-aminooxy-PEG-Methane, t-Boc-aminooxy-PEG-Propargyl, t-Boc-aminooxy-PEG-S-Ac, t-Boc-Aminooxy-PEG-Thiol, t-Boc-Aminooxy-PEG-Tos, Fmoc-aminooxy-PEG-acid, Trifluoroethyl-PEG-Aminooxy, Alkyne PEG (e.g.,endo-BCN-PEG, exo-BCN-PEG, Propargyl-PEG-acid, Propargyl-PEG-CH2CO2H, Propargyl-PEG-(CH2)3-acid, Propargyl-PEG-(CH2)3-methyl ester, Propargyl-PEG-Acrylate, Propargyl-PEG-alcohol, Propargyl-PEG-amine, Propargyl-PEG-methylamine (methylamine), Aminooxy-PEG-Propargyl, Propargyl-PEG-azide, Propargyl-PEG-bromide, Propargyl-PEG-Maleimide (maleimide), Propargyl-PEG-Ms, Propargyl-PEG-NHS ester, Propargyl-PEG-sulfonic acid, Propargyl-PEG-t-butyl ester, Propargyl-PEG-CH2CO2tBu, Propargyl-PEG-thiol, Propargyl-PEG-5-nitrophenyl carbonate (nitrophenyl carbonate), t-Boc-aminooxy-PEG-Propargyl, Bis-Propargyl-PEG, m-PEG-Propargyl), AzidoPEG (for example,Azido-PEG-acid, Azido-PEG-CH2CO2H, Azido-PEG-(CH2)3-methyl ester, Azido-PEG-Acrylate, Azido-PEG-alcohol, Azido-PEG-(CH2)3OH, Azido-PEG-amine, Azido-PEG-azide, Azido-PEG-Maleimide, Azido-PEG-methylamine, Azido-PEG-methylester, Azido-PEG-NHS ester, Azido-PEG-CH2CO2-NHS, Azido-PEG-oxazolidin-2-one, Azido-PEG-PFP ester, Azido-PEG-phosphonic acid, Azido-PEG-phosphonic acid ethyl ester, Azido-PEG-sulfonic acid, Azido-PEG-t-Boc-Hydrazide, Azido-PEG-t-butyl ester, Azido-PEG-CH2CO2-t-butyl ester, Azido-PEG-TFP ester, Azido-PEG-Tos, Aminooxy-PEG-azide, Bromo-PEG-azide, Bromoacetamido-PEG-azide, Carboxyrhodamine 110-PEG-Azide, Isothiocyanato-PEG-Azide, Isothiocyanato-PEG-Azide, m-PEG-azide, Propargyl-PEG-azide, TAMRA-PEG-Azide, t-Boc-N-Amido-PEG-Azide, t-Boc-Aminooxy-PEG-Azide, Thiol-PEG-Azide, Trifluoroethyl-PEG-Azide, Azido-PEG-amino acid, Azido-PEG4-4-nitrophenyl carbonate, S-Acetyl-PEG3-Azido, Azide, Trityl-PEG, 10-Azide), Alkyne PEG, DBCO-PEG, BCN-PEG, Propargyl-PEG, Bis-PEG-acid, Bis-PEG-NHS, Bis-PEG-PFP, Bis-Propargyl-PEG, Amine-PEG-Amine, Azido-PEG-azide, Bromo-PEG, Mal PEG.

[0195] In addition, as another embodiment, by using polyethylene glycol (PEG) described in WO2015 / 057699 and WO2017 / 165851, a conjugate that is expected to have better pharmacokinetics in vivo can be obtained.

[0196] Between anti-CAPRIN-1 antibodies and benzodiazepines The connectors between classes can be composed of a single type or multiple types.

[0197] As a method for producing anti-CAPRIN-1 antibodies and benzodiazepines The conjugate method comprises: using the ε-amino group of the lysine side chain of the antibody to bind benzodiazepine A method of combining with a cysteine ​​residue that forms a disulfide bond of an antibody by reducing the cysteine ​​residue to form a thiol group.

[0198] When using the ε-amino group of a lysine residue of an antibody, for example, a method of reacting an active ester (eg, N-hydroxysuccinimide ester) to form an amide bond can be used. In this case, since there are a large number of lysine residues in the antibody, the binding reacts non-specifically.

[0199] In the case of using the thiol group forming a disulfide bond on the side chain of the cysteine ​​of the antibody, a method can be used to generate a thiol group from the disulfide bond on the antibody using a reducing agent such as mercaptoethanol to react it with maleimide or α-haloamide. In addition, in order to stabilize the binding via the thiol group, there are methods such as using sulfone phenyl oxadiazole, 4-cyanoethynyloxy derivatives, etc. These combinations are more stable for a long time than the conjugate addition reaction of cysteine ​​to maleimide. In addition, the imide ring formed by the addition of thiol groups to maleimide is opened by hydrolysis, and once an amide bond is formed, the stability is improved, so a connector having an amino group near the imide group can also be used. In addition, there is also a thiol group of the cysteine ​​on the antibody to form a disulfide bond, and benzodiazepine is bound therebetween via two thiol groups. For example, a linker having two disulfide bond sites that can be generated by an amide group having two sulfones at the β position or dibromomaleimide can be used to form a cross-linking type bond.

[0200] The conjugate of the present invention can be prepared, for example, using THIOMAB (trademark) technology (see Nature Biotechnology 26, 925-932 (2008)), which is a method capable of introducing a predetermined number of thiol groups into a specific site of an antibody, or a method using ThioBridge (trademark) technology (see Methods Mol Biol. 2078: 113-129 (2020)), or RESPECT (trademark) technology (see MAbs. 9 (6): 907-915 (2017)).

[0201] The conjugate of the present invention can be prepared, for example, by reducing the antibody using a reducing agent, dithiothreitol (DTT), in a phosphate buffer to obtain an antibody having a thiol reactive group, thereby forming a conjugate with benzodiazepine. In addition to the method using a reducing agent, it can also be obtained by introducing Traut's Reagent (2-iminothiolane, N-succinimidyl S-acetylthioacetate (SATA)) to add a thiol group to the primary amine on the lysine residue of the antibody.

[0202] The amount of thiol groups attached to the antibody can be quantified, for example, by mixing a sample solution containing 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and an SH group with a phosphate buffer (pH 8.0) and distilled water, adding a DTNB solution dissolved in a phosphate buffer, Good's buffer or Tris buffer, incubating for a certain period of time, and then measuring the absorbance at 412 nm (see GLEllman, Arch. Biochem. Biophys., 82, 70 (1959)).

[0203] The thiol groups added after the disulfide bonds of the antibody are cleaved by the reduction treatment are preferably treated (blocked) to prevent the disulfide bonds from forming again. For example, N-ethylmaleimide (NEM) and iodoacetamide (2-iodoacetamide (IAA)) can be used for the blocking.

[0204] To use thiol-linked benzodiazepines attached to antibodies The conjugate can be formed by a known method. Specifically, for example, a linker reagent having a maleimide group or a linker reagent having a bromoacetamide group can be used as a linker reagent that specifically binds to the thiol group of the reduced antibody. For example, N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC) can be used as a linker having a maleimide group. In this case, by making benzodiazepine The amino group of SMCC forms an amide bond with the N-succinimidyl group of SMCC to obtain a conjugate.

[0205] As another embodiment, SMCC can be used to form an amide bond with the amino group on the activator, and then the thiol group attached to the antibody side can be bonded to the benzodiazepine-bound The maleimide group of SMCC was reacted to obtain a conjugate.

[0206] In another embodiment, two linkers may be used to form a bond between an antibody and a benzodiazepine. For example, a substance is prepared by forming an amide bond between the primary amino group present on the lysine residue on the antibody side and the N-succinimidyl group of SATA (N-succinimidyl-S-acetylthioacetate), adding a thiol group to the antibody, and conjugating SMCC with benzodiazepines. Benzodiazepines having an amino group on the side or to which an amino group is added by a conventional method The reaction was used to synthesize a substance that formed an amide bond with the N-succinimide group on SMCC, thereby combining benzodiazepines The maleimide group on the SMCC of the type 1 was reacted with the thiol group on the SATA bound to the antibody to obtain a conjugate.

[0207] As another embodiment, there can be mentioned a combination of an antibody and a benzodiazepine. In the preparation of conjugates of this type, for example, maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-Val-Cit-PAB) is used as a linker. mc-val-Cit-PAB (mc-vc-PAB) is a linker that can be cleaved by intracellular proteases (e.g., cathepsin B). DTT or the like is used to attach a thiol group to an antibody dissolved in a phosphate buffer in advance. On the other hand, a benzodiazepine having an amino group is made The benzodiazepine reacts with the benzyloxycarbonyl (PAB) on mc-Val-Cit-PAB to produce a benzodiazepine combined with mc-val-Cit-PAB. The conjugate can be obtained by reacting the above-mentioned antibody to which a thiol group is added.

[0208] As another embodiment, the method comprises: binding SATA to the primary amino group on the lysine residue of the antibody and adding a thiol group, and binding succinimidyl 3-(2-pyridyldithio) propionate (SPDP) to a benzodiazepine having an amino group. The N-succinimidyl group on SPDP forms an amide bond to obtain a conjugate.

[0209] The linker used in the present invention may be cut under intracellular conditions, and the benzodiazepine Benzodiazepines The substance with antitumor activity that is a part of the linker is free. For example, it is a linker that is cut by peptidases or proteases in cells. Preferably, it is a linker that is cut by lysosomes, endosome proteases, cathepsin B, cathepsin D, and plasmin. For example, a linker containing a polypeptide (Val-Cit, Phe-Leu or Gly-Phe-Leu-Gly) that can be cut by cathepsin B can be mentioned. In more detail, the linker described in U.S. Patent No. 6,214,345 can be used.

[0210] Furthermore, as another embodiment, a linker having glucuronic acid (preferably β-D-glucuronide) as described in WO2007 / 0711968 can be used as a linker for improving the blood stability, solubility and metabolism of the conjugate of the present invention and the benzodiazepine Alternatively, the methods described in WO2013 / 173337, WO2015 / 095755, WO2015 / 123679, and WO2018 / 031690 can be used.

[0211] Furthermore, as another embodiment, in order to make benzodiazepine Site-specific binding to an anti-CAPRIN-1 antibody can be achieved, for example, using the means described in WO2006 / 65533 and WO2018 / 160683.

[0212] In addition, as another embodiment, a benzodiazepine is combined with Conjugates of two or more drugs including the above class and anti-CAPRIN-1 antibodies can be obtained by the method described in WO2018 / 112253.

[0213] In order to obtain a product comprising the anti-CAPRIN-1 antibody of the present invention and benzodiazepine The composition of the conjugate can be obtained, for example, by separating a peak that is higher in molecular weight than the antibody before binding to the linker by gel filtration chromatography, etc. In order to detect the quality of the conjugate while maintaining the intact bivalent antibody, for example, the method described in WO2013 / 049410 can be used.

[0214] The anti-CAPRIN-1 antibody of the present invention and benzodiazepine Conjugates of the type with each antibody conjugated to a benzodiazepine The number of species can be quantified by a known method such as mass spectrometry, ELISA, electrophoresis, or chromatography such as HPLC.

[0215] <Antitumor effect of conjugate>

[0216] The anti-CAPRIN-1 antibody of the present invention and benzodiazepine The conjugates of the present invention have antitumor activity in vitro or in vivo. The so-called antitumor activity refers to the reduction, elimination, inhibition of proliferation, apoptosis, cell necrosis or killing of the targeted cancer cells. The antitumor effect of the conjugates of the present invention can be known by investigating the antitumor activity against cancer.

[0217] In order to know the anti-tumor effect in vivo, the conjugate can be administered to an organism with cancer, the size of the tumor after administration can be measured, and the size of the cancer can be investigated over time to evaluate. The anti-tumor effect of the present invention can also be evaluated by investigating the survival rate. In addition, it can also be evaluated by investigating the production capacity of cytokines or chemokines. The anti-tumor effect of the conjugate of the present invention can be further known by investigating the prevention of cancer, the prevention of metastasis, or the prevention of recurrence.

[0218] The higher the binding affinity of the conjugate of the present invention to the CAPRIN-1 protein on the surface of cancer cells, the stronger the anti-tumor effect can be expected. The binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5×10 8 M -1 , at least 10 9 M -1 , at least 5×10 9 M -1 , at least 10 10 M -1 , at least 5×10 10 M -1 , at least 10 11 M -1 , at least 5×10 11 M -1 , at least 10 12 M -1 , or at least 10 13 M -1 .

[0219] The ability of the conjugate of the present invention to bind to CAPRIN-1 can be identified by a binding assay using, for example, the surface plasmon resonance method (SPR method), ELISA method, Western blotting method, immunofluorescence, flow cytometry, and the like.

[0220] As described above, the conjugate of the present invention has an enhanced antitumor effect compared to the anti-CAPRIN-1 antibody alone, and the enhancement rate is preferably 30% or more, more preferably 40% or more, further preferably 50% or more, further more preferably 55% or more, further more preferably 60% or more, further more preferably 65% ​​or more, and most preferably 70% or more. The enhancement rate of the antitumor effect of the conjugate of the present invention relative to the anti-CAPRIN-1 antibody alone can be calculated by administering the effective amount to cancer-bearing mice under the same conditions and comparing the tumor volume on the 10th day after the start of administration.

[0221] <Pharmaceutical composition, method for treating and / or preventing cancer>

[0222] The target of the pharmaceutical composition for treating and / or preventing cancer of the present invention is not particularly limited as long as it is a cancer (cell) expressing the CAPRIN-1 protein.

[0223] As used herein, the terms "tumor" and "cancer" refer to malignant neoplasms and are used interchangeably.

[0224] The cancer targeted in the present invention may be any cancer as long as it expresses CAPRIN-1 protein on the cell membrane surface. Preferred are breast cancer, kidney cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma. Testicular cancer, thyroid cancer, head and neck cancer, urothelial carcinoma.

[0225] The cancer more specifically includes, for example, breast cancer, combined breast cancer, malignant mixed tumor of the breast, intraductal papillary adenocarcinoma, recurrent metastatic breast cancer, lung adenocarcinoma, non-small cell lung cancer (NSCLC), squamous cell non-small cell lung cancer, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, glioma as a neuroepithelial tissue tumor, glioblastoma, neuroblastoma, ependymoma, neuronal tumor, fetal neuroectodermal tumor, schwannoma, neurofibroma, meningioma, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, digestive organ lymphoma, small to medium cell lymphoma, cecal cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, sigmoid colon cancer, rectal cancer, ovarian epithelial cancer, germ cell tumor, stromal cell tumor, pancreatic duct cancer, invasive pancreatic duct cancer, pancreatic cancer adenocarcinoma, metastatic adenocarcinoma, adenocarcinoma Follicular cell carcinoma, adenosquamous cell carcinoma, giant cell tumor, intraductal papillary mucinous neoplasm of the pancreas, mucinous cystadenocarcinoma, pancreatoblastoma, pancreatic head cell tumor, Frants tumor, serous cystadenocarcinoma, solid papillary carcinoma, gastrinoma, glucagonoma, insulinoma, multiple endocrine neoplasia syndrome 1 (Wermer syndrome), non-functional islet cell tumor, somatostatinoma, VIP producing tumor, cervical cancer, uterine body cancer, fibrosarcoma, bone and joint sarcoma, Ewing sarcoma, Wilms' tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, soft tissue malignant tumor, teratocarcinoma, head and neck cancer including hypopharyngeal cancer, middle pharyngeal cancer, tongue cancer, upper pharyngeal cancer, oral cancer, lip cancer, sinus cancer, laryngeal cancer, recurrent brain tumor, etc., but not limited to these.

[0226] In addition, preferred subjects (patients) are mammals, such as primates, pets, livestock, and competitive animals, and humans, dogs, and cats are particularly preferred.

[0227] When the conjugate used in the present invention is used as a pharmaceutical composition, it can be formulated using methods known to those skilled in the art. For example, it can be used parenterally in the form of a sterile solution or suspension injection with water or a pharmaceutically acceptable liquid other than water. For example, it is considered that it can be appropriately combined with a pharmacologically acceptable carrier or medium, specifically sterile water and / or physiological saline, vegetable oil, emulsifier, suspending agent, surfactant, stabilizer, fragrance, excipient, binder, etc., and formulated by mixing in a unit dosage manner required for generally believed pharmaceutical implementation. The amount of active ingredient in these preparations is such an amount that an appropriate dosage within the indicated range can be obtained.

[0228] When the conjugate of the present invention is used as a pharmaceutical composition, it may contain any salt, surfactant, buffer, sugar, and cryoprotectant (including a part of sugar) and be formulated in a freeze-dried state.

[0229] The sterile composition for injection can use a medium such as distilled water for injection, and is prescribed according to the usual formulation implementation. As an aqueous solution for injection, for example, physiological saline, isotonic solution containing glucose and / or other adjuvants, such as D-sorbitol, D-mannose, D-mannitol, and sodium chloride can be cited, and it can also be used in conjunction with a suitable dissolution aid, such as alcohol, specifically ethanol, polyols such as propylene glycol, polyethylene glycol, nonionic surfactants such as polysorbate 80 (TM), HCO-60. As an oily liquid, sesame oil and soybean oil can be cited, and it can also be used in conjunction with benzyl benzoate and benzyl alcohol as dissolution aids. In addition, it can also be mixed with buffers such as phosphate buffer, sodium acetate buffer, analgesics such as procaine hydrochloride, stabilizers such as benzyl alcohol, phenol, and antioxidants. The prepared injection is usually filled in a suitable ampoule. As an oily liquid, sesame oil and soybean oil can be cited, and it can also be used in conjunction with benzyl benzoate and benzyl alcohol as dissolution aids. In addition, it can also be mixed with a buffer such as phosphate buffer, sodium acetate buffer, an analgesic such as procaine hydrochloride, a stabilizer such as benzyl alcohol, phenol, and an antioxidant. The prepared injection is usually filled into a suitable ampoule.

[0230] Administration is oral or parenteral, preferably parenteral, and specifically includes injection, nasal administration, pulmonary administration, transdermal administration, etc. Examples of injections include systemic or local administration by, for example, intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, intratumor injection, etc.

[0231] In addition, an appropriate administration method can be selected according to the patient's age, weight, sex, symptoms, etc. The administration amount of the pharmaceutical composition containing the antibody or the polynucleotide encoding the antibody can be, for example, 0.5 mg, 1 mg, 2 mg, 3 mg, 5 mg, 10 mg, 20 mg, 50 mg, 75 mg, 100 mg, 200 mg, 500 mg or 1000 mg per kg of body weight in the range of 0.0001 mg to 1000 mg per kg of body weight. Or, for example, the administration amount can be selected in the range of 0.001 to 100000 mg / individual per patient, but it is not necessarily limited to these values.

[0232] The dosage and administration method vary depending on the patient's weight, age, sex, symptoms, etc., and can be appropriately selected by those skilled in the art.

[0233] By administering a pharmaceutical composition for the treatment and / or prevention of cancer comprising the conjugate of the present invention as an active ingredient to a subject, the aforementioned cancers expressing CAPRIN-1 on the cell membrane surface, preferably breast cancer, kidney cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, head and neck cancer, and urothelial carcinoma can be treated and / or prevented.

[0234] Example

[0235] Hereinafter, the present invention will be specifically described based on Examples, but the scope of the present invention is not limited to these specific examples.

[0236] (Example 1) Anti-CAPRIN-1 Polyclonal Antibody

[0237] The anti-CAPRIN-1 polyclonal antibody immunoreactive with the CAPRIN-1 protein used in the conjugate of the present invention is prepared by mixing 1 mg of the human CAPRIN-1 recombinant protein shown in sequence number 2 and sequence number 4 prepared in Example 3 of WO2010 / 016526 with an equal volume of Freund's incomplete adjuvant (IFA) solution, and administering it subcutaneously to rabbits every 2 weeks for a total of 4 times. Blood is then collected to obtain antiserum containing polyclonal antibodies. The obtained antiserum is purified using a protein G carrier (GE Healthcare Biosciences) to obtain a polyclonal antibody against the CAPRIN-1 protein (anti-CAPRIN-1 polyclonal antibody #1). In addition, the serum of a rabbit to which the antigen was not administered was purified using a protein G carrier in the same manner as described above as a rabbit control antibody.

[0238] In addition, the following polyclonal antibodies #2 to #6 against partial polypeptides of CAPRIN-1 were obtained by the same method as the method for preparing the above-mentioned polyclonal antibody against CAPRIN-1 protein.

[0239] SEQ ID NO: CAPRIN-1 protein Anti-CAPRIN-1 polyclonal antibody #2 directed against a partial polypeptide represented by SEQ ID NO: 37 described in WO2011 / 096528 (SEQ ID NO: 31 in the present specification).

[0240] Anti-CAPRIN-1 polyclonal antibody #3 directed against the partial polypeptide represented by SEQ ID NO: 5 described in WO2013 / 018894 (SEQ ID NO: 32 in the present specification).

[0241] Anti-CAPRIN-1 polyclonal antibody #4 directed against a partial polypeptide represented by SEQ ID NO: 5 described in WO2013 / 125654 (SEQ ID NO: 33 in the present specification).

[0242] Anti-CAPRIN-1 polyclonal antibody #5 directed against a partial polypeptide represented by SEQ ID NO: 37 described in WO2011 / 096533 (SEQ ID NO: 34 in the present specification).

[0243] Anti-CAPRIN-1 polyclonal antibody #6 directed against a partial polypeptide represented by SEQ ID NO: 37 described in WO2011 / 096534 (SEQ ID NO: 35 in the present specification).

[0244] (Example 2) Anti-CAPRIN-1 Monoclonal Antibody

[0245] The anti-CAPRIN-1 monoclonal antibodies used in the conjugate of the present invention are the following monoclonal antibodies.

[0246] As a monoclonal antibody against CAPRIN-1 described in WO2011 / 096528, an antibody in which CDR1 to 3 of the heavy chain variable region are composed of the amino acid sequences of SEQ ID NOs: 36, 37 and 38, respectively, and CDR1 to 3 of the light chain variable region are composed of SEQ ID NOs: 40, 41 and 42, respectively (for example, an antibody comprising the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 39 including the CDR1 to 3 of the heavy chain variable region, and the amino acid sequence of the light chain variable region represented by SEQ ID NO: 43 including the CDR1 to 3 of the light chain variable region).

[0247] As a monoclonal antibody against CAPRIN-1 described in WO2015 / 020212, an antibody in which CDR1 to 3 of the heavy chain variable region are composed of the amino acid sequences of SEQ ID NOs: 44, 45 and 46, respectively, and CDR1 to 3 of the light chain variable region are composed of SEQ ID NOs: 48, 49 and 50, respectively (for example, an antibody comprising the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 47 including the CDR1 to 3 of the heavy chain variable region, and the amino acid sequence of the light chain variable region represented by SEQ ID NO: 51 including the CDR1 to 3 of the light chain variable region).

[0248] As a monoclonal antibody against CAPRIN-1 described in WO2011 / 096519, an antibody in which CDR1 to 3 of the heavy chain variable region are composed of the amino acid sequences of SEQ ID NOs: 52, 53 and 54, respectively, and CDR1 to 3 of the light chain variable region are composed of SEQ ID NOs: 56, 57 and 58, respectively (for example, an antibody comprising the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 55 including the CDR1 to 3 of the heavy chain variable region, and the amino acid sequence of the light chain variable region represented by SEQ ID NO: 59 including the CDR1 to 3 of the light chain variable region).

[0249] As a monoclonal antibody against CAPRIN-1 described in WO2013 / 125654, an antibody in which CDR1 to 3 of the heavy chain variable region are composed of the amino acid sequences of SEQ ID NOs: 60, 61 and 62, respectively, and CDR1 to 3 of the light chain variable region are composed of SEQ ID NOs: 64, 65 and 66, respectively (for example, an antibody comprising the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 63 including the CDR1 to 3 of the heavy chain variable region, and the amino acid sequence of the light chain variable region represented by SEQ ID NO: 67 including the CDR1 to 3 of the light chain variable region).

[0250] The monoclonal antibody against CAPRIN-1 described in WO2011 / 096517 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:68 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:69.

[0251] The monoclonal antibodies against CAPRIN-1 described in WO2011 / 096528 include an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 70, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 71; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 72, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 73; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 74, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 75; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 76, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 77; and an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 78, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 79.

[0252] As the monoclonal antibody against CAPRIN-1 described in WO2011 / 096533, an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 80, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 81; an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 82, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 83.

[0253] As the monoclonal antibody against CAPRIN-1 described in WO2011 / 096534, an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 84, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 85; an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 86, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 87.

[0254] As monoclonal antibodies against CAPRIN-1 described in WO2010 / 016526, there are antibodies comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 88, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 89; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 90, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 91; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 92, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 93; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 94 The invention relates to an antibody comprising the amino acid sequence of a heavy chain variable region shown in the amino acid sequence of sequence number 96 and the amino acid sequence of a light chain variable region shown in the amino acid sequence of sequence number 97; an antibody comprising the amino acid sequence of a heavy chain variable region shown in the amino acid sequence of sequence number 98 and the amino acid sequence of a light chain variable region shown in the amino acid sequence of sequence number 99; and an antibody comprising the amino acid sequence of a heavy chain variable region shown in the amino acid sequence of sequence number 100 and the amino acid sequence of a light chain variable region shown in the amino acid sequence of sequence number 101.

[0255] As the monoclonal antibody against CAPRIN-1 described in WO2013 / 018894, an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 102, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 103; an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 104, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 105.

[0256] The monoclonal antibody against CAPRIN-1 described in WO2013 / 018892 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:106 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:107.

[0257] The monoclonal antibody against CAPRIN-1 described in WO2013 / 018891 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:108 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:109.

[0258] The monoclonal antibody against CAPRIN-1 described in WO2013 / 018889 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 111.

[0259] The monoclonal antibody against CAPRIN-1 described in WO2013 / 018883 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:112 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:113.

[0260] The monoclonal antibody against CAPRIN-1 described in WO2013 / 125636 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:114 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:115.

[0261] As the monoclonal antibody against CAPRIN-1 described in WO2013 / 125654, an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 116, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 117; an antibody comprising the amino acid sequence of the heavy chain variable region shown by the amino acid sequence of SEQ ID NO: 118, and the amino acid sequence of the light chain variable region shown by the amino acid sequence of SEQ ID NO: 119.

[0262] The monoclonal antibody against CAPRIN-1 described in WO2013 / 125630 includes an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 121.

[0263] The monoclonal antibodies against CAPRIN-1 described in WO2015 / 020212 include an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 122, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 123; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 124, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 125; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 126, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 127; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 128, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 129; an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 130, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 131; and an antibody comprising an amino acid sequence of a heavy chain variable region as shown in the amino acid sequence of SEQ ID NO: 132, and an amino acid sequence of a light chain variable region as shown in the amino acid sequence of SEQ ID NO: 133.

[0264] The base sequence was designed so that the heavy chain variable region CDR1 to 3 of the anti-CAPRIN-1 monoclonal antibody described above, which consisted of the amino acid sequences of SEQ ID NO: 36, SEQ ID NO: 37, and SEQ ID NO: 38, respectively, and the framework region contained the sequence of a human antibody, could be expressed, and this was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted. Similarly, the base sequence was designed so that the light chain variable region CDR1 to 3 consisted of the amino acid sequences of SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42, respectively, and the framework region contained the sequence of a human antibody, and this was inserted into a mammalian expression vector into which the light chain constant region of human IgG1 had been inserted. The above two recombinant expression vectors were introduced into mammalian cells according to conventional methods to obtain the culture supernatant of humanized monoclonal antibody #1 (humanized antibody #1) against CAPRIN-1, wherein CDR1 to 3 of the heavy chain variable region consist of the amino acid sequences of sequence numbers 36, 37 and 38, respectively, and CDR1 to 3 of the light chain variable region consist of sequence numbers 40, 41 and 42, respectively.

[0265] Similarly, the base sequence of the heavy chain variable region shown in sequence number 47 is designed so that CDR1 to 3 of the heavy chain variable region are composed of the amino acid sequences of sequence numbers 44, 45 and 46, respectively, and the framework region contains the sequence of a human antibody, and is inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 has been inserted. Similarly, the base sequence was designed in such a way that CDR1 to 3 of the light chain variable region consisted of the amino acid sequences of SEQ ID NOs: 48, 49 and 50, respectively, and the framework region contained the sequence of a human antibody in the heavy chain variable region shown in SEQ ID NO: 51, and the base sequence was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted. The above two recombinant expression vectors were introduced into mammalian cells according to a conventional method, and the culture supernatant of humanized anti-CAPRIN-1 monoclonal antibody #2 (humanized antibody #2) containing CDR1 to 3 of the heavy chain variable region consisting of the amino acid sequences of SEQ ID NOs: 44, 45 and 46, and CDR1 to 3 of the light chain variable region consisting of the amino acid sequences of SEQ ID NOs: 48, 49 and 50, respectively, was obtained.

[0266] Similarly, the culture supernatant of humanized anti-CAPRIN-1 monoclonal antibody #3 (humanized antibody #3) was obtained, wherein CDR1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO: 52, SEQ ID NO: 53, and SEQ ID NO: 54, respectively, and CDR1 to 3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO: 56, SEQ ID NO: 57, and SEQ ID NO: 58, respectively.

[0267] Similarly, the culture supernatant of humanized anti-CAPRIN-1 monoclonal antibody #4 (humanized antibody #4) was obtained, wherein CDR1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO: 60, SEQ ID NO: 61, and SEQ ID NO: 62, respectively, and CDR1 to 3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO: 64, SEQ ID NO: 65, and SEQ ID NO: 66, respectively.

[0268] Similarly, culture supernatants containing the following humanized anti-CAPRIN-1 monoclonal antibodies #9 to #41 (humanized antibodies #9 to #41) were obtained.

[0269] A humanized monoclonal antibody #9 (humanized antibody #9) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 68 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 69.

[0270] A humanized monoclonal antibody #10 (humanized antibody #10) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 70 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 71.

[0271] A humanized monoclonal antibody #11 (humanized antibody #11) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 72 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 73.

[0272] Humanized monoclonal antibody #12 (humanized antibody #12) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 75.

[0273] Humanized monoclonal antibody #13 (humanized antibody #13) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 76 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 77.

[0274] Humanized monoclonal antibody #14 (humanized antibody #14) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 78 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 79.

[0275] A humanized monoclonal antibody #15 (humanized antibody #15) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:80 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:81.

[0276] A humanized monoclonal antibody #16 (humanized antibody #16) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 82 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 83.

[0277] A humanized monoclonal antibody #17 (humanized antibody #17) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 84 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 85.

[0278] A humanized monoclonal antibody #18 (humanized antibody #18) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 86 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 87.

[0279] An antibody humanized monoclonal antibody #19 (humanized antibody #19) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 88 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 89.

[0280] A humanized monoclonal antibody #20 (humanized antibody #20) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:90 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:91.

[0281] A humanized monoclonal antibody #21 (humanized antibody #21) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:92 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:93.

[0282] A humanized monoclonal antibody #22 (humanized antibody #22) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:94 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:95.

[0283] A humanized monoclonal antibody #23 (humanized antibody #23) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:96 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:97.

[0284] A humanized monoclonal antibody #24 (humanized antibody #24) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:98 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:99.

[0285] A humanized monoclonal antibody #25 (humanized antibody #25) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:100 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:101.

[0286] A humanized monoclonal antibody #26 (humanized antibody #26) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:102 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:103.

[0287] A humanized monoclonal antibody #27 (humanized antibody #27) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:104 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:105.

[0288] A humanized monoclonal antibody #28 (humanized antibody #28) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO:106 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO:107.

[0289] A humanized monoclonal antibody #29 (humanized antibody #29) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 108 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 109.

[0290] Humanized monoclonal antibody #30 (humanized antibody #30) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 111.

[0291] A humanized monoclonal antibody #31 (humanized antibody #31) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 113.

[0292] A humanized monoclonal antibody #32 (humanized antibody #32) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 115.

[0293] A humanized monoclonal antibody #33 (humanized antibody #33) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 117.

[0294] A humanized monoclonal antibody #34 (humanized antibody #34) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 118 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 119.

[0295] A humanized monoclonal antibody #35 (humanized antibody #35) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 121.

[0296] A humanized monoclonal antibody #36 (humanized antibody #36) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 122 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 123.

[0297] A humanized monoclonal antibody #37 (humanized antibody #37) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 124 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 125.

[0298] A humanized monoclonal antibody #38 (humanized antibody #38) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 127.

[0299] A humanized monoclonal antibody #39 (humanized antibody #39) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 128 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 129.

[0300] A humanized monoclonal antibody #40 (humanized antibody #40) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 131.

[0301] A humanized monoclonal antibody #41 (humanized antibody #41) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 132 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 133.

[0302] In addition, for the humanized antibody #1 among the anti-CAPRIN-1 monoclonal antibodies, the base sequence was designed so that the heavy chain variable region CDR1 to 3 consisted of the amino acid sequences of SEQ ID NO: 36, SEQ ID NO: 37, and SEQ ID NO: 38, respectively, and the framework region contained the sequence of a human antibody, and was introduced into a mammalian expression vector into which the heavy chain constant region of human IgG1 in which the amino acid serine (Ser) at position 239 of EU index number was replaced by aspartic acid (Asp), and the amino acid isoleucine (Ile) at position 332 of EU index number was replaced by glutamic acid (Glu) was inserted. In addition, the base sequence was designed so that the amino acid sequence of the light chain variable region in which the CDR1 to 3 of the light chain variable region consisted of SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42, respectively, and the framework region contained the sequence of a human antibody could be expressed, and was inserted into a mammalian expression vector into which the light chain constant region of human IgG1 was inserted. The above two recombinant expression vectors were introduced into mammalian cells according to conventional methods to obtain the culture supernatant of humanized monoclonal antibody #5 (humanized antibody #5) against CAPRIN-1, which comprises a heavy chain full-length amino acid sequence consisting of the heavy chain variable region produced above and a heavy chain constant region of human IgG1 in which the amino acid serine (Ser) at EU number 239 is replaced by aspartic acid (Asp) and the amino acid isoleucine (Ile) at EU number 332 is replaced by glutamic acid (Glu), and a light chain full-length amino acid sequence consisting of the light chain variable region produced above and a human light chain constant region.

[0303] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #6 (humanized antibody #6) containing the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region of the humanized antibody #2 prepared above was obtained.

[0304] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #7 (humanized antibody #7) containing the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region of the humanized antibody #3 prepared above was obtained.

[0305] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #8 (humanized antibody #8) containing the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region of the humanized antibody #4 prepared above was obtained.

[0306] Similarly, culture supernatants of humanized anti-CAPRIN-1 antibodies #42 to #74 (humanized antibodies #42 to #74) containing the amino acid sequences of the heavy chain variable regions and light chain variable regions of humanized antibodies #9 to #41 prepared above were obtained.

[0307] The obtained culture supernatants containing humanized anti-CAPRIN-1 monoclonal antibodies #1 to #74 were purified using Hitrap Protein A Sepharose FF (GE Healthcare) according to a conventional method, replaced with PBS (-), and filtered through a 0.22 μm filter (Millipore).

[0308] (Example 3) Preparation of a conjugate of an anti-CAPRIN-1 antibody and a PBD derivative (SGD-1910)

[0309] Conjugates of the anti-CAPRIN-1 polyclonal antibodies #1 to #6 described in Example 1 and SGD-1910 (CAS #1342820-51-2) were prepared according to conventional methods. A solution was prepared by dissolving the purified anti-CAPRIN-1 polyclonal antibody #1 in PBS (-) at a concentration of 10 mg / ml. A PBS (-) solution containing 100 mM EDTA and 50 mM L-cysteine ​​was added to the solution containing the polyclonal antibody #1 at a ratio of 1 / 10, and then a reduction reaction was performed at 4°C for 1 hour. After reduction, the solution was replaced with PBS (-) containing 1 mM EDTA using a Zeba (trademark) Spin Desalting Column (MWCO70k or 40k), and the polyclonal antibody was allowed to stand at room temperature for 18 to 72 hours to reoxidize. Propylene glycol was added to the reoxidized polyclonal antibody #1 to make it 50%. 10 mM SGD-1910 dissolved in DMSO was added to the solution containing the polyclonal antibody #1 in a molar ratio of 1:5 between the antibody and SDG-1910, and the reaction was allowed to proceed for more than 1 hour at room temperature. After the reaction, the SGD-1910 not bound to the antibody was removed using a Zeba (trademark) Spin Desalting Column (MWCO70k or 40k), and the solvent was replaced with PBS (-) (implemented twice). After the buffer was replaced with PBS (-), a 0.22 μm sterilization filter membrane was used for filtration to obtain a solution containing the anti-CAPRIN-1 polyclonal antibody #1-SDG-1910 (conjugate 1) of the present invention. Solutions containing conjugates using SDG-1910 were obtained similarly for anti-CAPRIN-1 polyclonal antibodies #2 to #6 (the conjugate using anti-CAPRIN-1 polyclonal antibody #2: conjugate 2; the conjugate using anti-CAPRIN-1 polyclonal antibody #3: conjugate 3; the conjugate using anti-CAPRIN-1 polyclonal antibody #4: conjugate 4; the conjugate using anti-CAPRIN-1 polyclonal antibody #5: conjugate 5; the conjugate using anti-CAPRIN-1 polyclonal antibody #6: conjugate 6). In addition, a solution containing a conjugate (conjugate 10) with SDG-1910 was obtained using the humanized antibody #4 as the anti-CAPRIN-1 monoclonal antibody described in Example 2 by the same method as above. A solution containing a conjugate (control conjugate 1) with a linker of SDG-1910 was obtained by the same method as above for the rabbit control antibody described in Example 1 that does not react with the CAPRIN-1 protein.

[0310] Furthermore, a conjugate of humanized antibody #1 and SGD-1910 (CAS#1342820-51-2) as the anti-CAPRIN-1 monoclonal antibody described in Example 2 was prepared according to a conventional method. A solution was prepared by dissolving the purified humanized antibody #1 in PBS(-) at a concentration of 10 mg / mL. A PBS(-) solution containing TCEP at a final concentration of 10 mM was added to the solution containing humanized antibody #1, and then a reduction reaction was performed at room temperature for 1 hour. After reduction, the solution was replaced with PBS(-) containing 1 mM EDTA using a Zeba (trademark) Spin Desalting Column (MWCO70k or 40k), and then an equal amount of propylene glycol was added. To the solution containing humanized antibody #1, 10 mM SGD-1910 dissolved in DMSO was added in a molar ratio of antibody to SDG-1910 of 1:3, and it was reacted at room temperature for more than 1 hour. After the reaction, the SGD-1910 that was not bound to the antibody was removed using a Zeba (trademark) Spin Desalting Column (MWCO70k or 40k), and the solvent was replaced with PBS (-) (implemented twice). After the buffer was replaced with PBS (-), filtration was performed using a 0.22 μm sterilization filter membrane to obtain a solution containing the humanized antibody #1-SDG-1910 (conjugate 7) of the present invention. Similarly, for anti-CAPRIN-1 polyclonal antibodies #2 to #6 and humanized antibodies #2, #3, #5 to #74, solutions containing conjugates using SDG-1910 were also obtained (conjugate 8, conjugate 9, conjugate 11 to conjugate 53). By the same operation as above, a solution containing a conjugate with a linker to SDG-1910 (control conjugate 2) was obtained using the same method for a human IgG control antibody that does not react with the CAPRIN-1 protein.

[0311] (Example 4) Specific reactivity of the conjugate to CAPRIN-1 protein and CAPRIN-1 expressing cancer cells

[0312] The specific reactivity of the (anti-CAPRIN-1 antibody-SDG-1910 conjugate) prepared in Example 3 to the CAPRIN-1 protein and whether it showed reactivity to the cell membrane surface of human cancer cells and mouse cancer cells expressing the CAPRIN-1 protein were evaluated.

[0313] The specific reactivity to CAPRIN-1 protein was confirmed using the ELISA method. 100 μL of a 1 μg / mL CAPRIN-1 protein solution was added to each well of a 96-well plate in advance and allowed to stand at 4°C for 18 hours. After washing each well three times with PBS-T, 400 μL of a 0.5% bovine serum albumin (BSA) solution was added to each well and allowed to stand at room temperature for 3 hours. The solution was removed, and each well was washed three times with 400 μL of PBS-T. Then, 100 μL of a solution containing conjugates 1 to 6 and control conjugate 1 was added to each well and allowed to stand at room temperature for 2 hours. After washing each well three times with PBS-T, 100 μL of an HRP-labeled anti-rabbit IgG antibody diluted 5000 times with PBS was added to each well and allowed to stand at room temperature for 1 hour. After washing the wells 3 times with PBS-T, 100 μL of TMB substrate solution was added to each well and allowed to stand for 15 to 30 minutes to perform a color reaction. After color development, 100 μL of 1N sulfuric acid was added to each well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorbance meter. As a result, it was confirmed that conjugates 1 to 6 had higher absorbance values ​​than the negative control conjugate 1, and specifically reacted to the CAPRIN-1 protein.

[0314] Similarly, the specific reaction to CAPRIN-1 protein was detected by ELISA using HRP-labeled anti-human antibody for the anti-CAPRIN-1 antibody-SDG-1910 conjugates using humanized antibodies #1 to #74 prepared in Example 3. The results confirmed that the absorbance value was higher than that of human IgG (SIGMA) used as a negative control and control conjugate 2, indicating that the conjugates specifically reacted with the CAPRIN-1 protein.

[0315] Next, the reactivity to the cell membrane surface of cancer cells expressing CAPRIN-1 was confirmed by flow cytometry. 5Human breast cancer cells BT-474 (obtained from ATCC) and mouse breast cancer cells 4T1 (obtained from ATCC) were centrifuged in 1.5 ml microcentrifuge tubes, and 100 μL of a solution containing conjugates 1 to 6 and control conjugate 1 was added to each tube and allowed to stand at 4° C. for 1 hour. After washing with PBS, Alexa488-labeled anti-rabbit IgG (H+L) diluted 100-fold with PBS (-) containing 0.5% FBS (0.5% FBS-PBS (-)) was added and allowed to stand at 4° C. for 1 hour. After washing with 0.5% FBS-PBS (-), the cells were reacted with BD Horizon Fixable Viability Stain (FVS) Reagents (FVS450, Biotin / Dikinson ADC Pani) to stain dead cells, and then the fluorescence intensity was measured using FACSFortessa (trademark) (biotin / Dikinson ADC Pani). As a result, it was confirmed that the anti-CAPRIN-1 antibody-SDG-1910 conjugates (conjugates 1 to 6) had higher fluorescence intensity than the negative control control conjugate 1, that is, they reacted more strongly with the cell surface of human cancer cell BT474 expressing CAPRIN-1.

[0316] Similarly, the specific reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates using humanized antibodies #1 to #74 to human cancer cells BT474 and mouse cancer cells 4T1, which are CAPRIN-1 expressing cancer cells, was detected using Alexa 488 labeled anti-human IgG (H+L) antibodies. As a result, it was confirmed that the anti-CAPRIN-1 antibody-SDG-1910 conjugates using humanized antibodies #1 to #74 had higher fluorescence intensity than the control conjugate 2 as a negative control, that is, they reacted more strongly with the cell surfaces of human cancer cells BT474 and 4T1 expressing CAPRIN-1.

[0317] Similarly, the reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates prepared in Example 3 using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. The reactivity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates to various human cancer cells and mouse cancer cells was confirmed. (KKU213), fibrosarcoma cells (HT-1080), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB), melanoma cells (Malme-3M), head and neck cancer cells (FaDu), mouse renal cancer cells (Renca) in which the expression of the CAPRIN-1 gene was confirmed, and mouse breast cancer cells (4T1). The anti-CAPRIN-1 antibody-SDG-1910 conjugate prepared in Example 3 was confirmed to have stronger fluorescence intensity than the control conjugate 1 and control conjugate 2 as negative controls, and reacted more strongly with the cell membrane surface of cancer cells expressing CAPRIN-1.

[0318] (Example 5) Internalization

[0319] The internalization of the anti-CAPRIN-1 antibody-SDG-1910 conjugates prepared in Example 3 using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 into cancer cells was evaluated in vitro. 4 μg / mL of the anti-CAPRIN-1 antibody-SDG-1910 conjugate and 2 μg / mL of Incucyte (trademark) Human Fabfluor-pH Antibody Labeling Dye were mixed in equal amounts and allowed to stand at 37°C for 1 hour to form a complex of the anti-CAPRIN-1 antibody-SDG-1910 conjugate and Human Fabfluor-pH Antibody Labeling Dye. 100 μL of the complex was added to 2×10 5The cells were reacted at 37°C for 6 hours in HT29 cancer cells. After the reaction, the internalization was evaluated by measuring the fluorescence intensity by flow cytometry according to conventional methods. The evaluation results showed that the anti-CAPRIN-1 antibody-SDG-1910 conjugate showed a stronger fluorescence intensity than the fluorescence intensity of the negative control (HT29 in which only the anti-CAPRIN-1 antibody-SDG-1910 conjugate was reacted). This result shows that the anti-CAPRIN-1 antibody-SDG-1910 conjugate is internalized into human cancer cells. Similarly, the internalization of trastuzumab, an antibody targeting HER2, cetuximab, an antibody targeting EGFR, enfortumab vedotin, an ADC targeting nectin-4, and sacituzumab govitecan, an ADC targeting TROP2, into HT29 was evaluated by flow cytometry, and the results showed that the fluorescence intensity of the anti-CAPRIN-1 antibody-SDG-1910 conjugate was stronger than that of any of the antibodies.

[0320] (Example 6) Antitumor activity of the conjugate

[0321] The anti-tumor activity of the anti-CAPRIN-1 antibody-SDG-1910 conjugates using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 against cancer cells was evaluated in vitro. The anti-tumor activity was evaluated against breast cancer cells (BT-474), colorectal cancer cells (HT-29, HCT116), gastric cancer cells (NCI-N87), uterine cancer cells (Ishikawa), pancreatic cancer cells (Panc10.5), ovarian cancer cells (MCAS, TOV112D), and head and neck cancer cells (FaDu), which are human cancer cells whose expression of CAPRIN-1 on the cell membrane surface of cancer cells was confirmed in Example 4.

[0322] The cancer cells cultured by a standard method were inoculated in a black 96-well plate capable of cell culture (96well optical Btm Plt PolymerBase Black w / Lid Cell Culture SterilePS165305, Thermo) at 90 μL / well at 1 to 4 × 10 cells / well. 4 , and cultured in an incubator at 5% CO2 and 37°C for 18 to 24 hours. -2 μg / mL~10 210 μL of the anti-CAPRIN-1 antibody-SDG-1910 conjugate using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 was added to each well and incubated in an incubator at 5% CO2 and 37°C. Cell viability was measured from 3 days to 4 days after the start of incubation using Cell Titer GlowLuminescent Cell Viability Assay (#7573, Promega). 100 μL of substrate was added to each well, shaken for 2 minutes using a microplate shaker, and after standing for 10 minutes, the luminescent signal intensity was measured using SpectraMax (registered trademark) iD3 (MOLECULAR DEVICE) to calculate the ATP content of the surviving cells.

[0323] As a result, when the number of cancer cells surviving in the wells without the conjugate added was taken as 100%, the number of cancer cells surviving in the anti-CAPRIN-1 antibody-SDG-1910 conjugates using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 described in the present invention was less than 70% in the concentration range of 1 μg / mL or more. The above results indicate that the conjugate of the antibody against CAPRIN-1 and PBD exhibits antitumor activity against cancer cells.

[0324] (Example 7) Antitumor effect of the conjugate in cancer-bearing mice

[0325] The anti-tumor effects of conjugates 7 and 10, which are anti-CAPRIN-1 antibody-SDG-1910 conjugates prepared in Example 3, were evaluated in vivo in cancer-bearing mice. The anti-tumor effects of the conjugates of the present invention were studied using NOD-SCID mice transplanted with human cancer cells expressing CAPRIN-1 protein. 7 Individual colorectal cancer cells HT29 were mixed with Matrigel (SIGMA) and transplanted subcutaneously to grow to a tumor size of 50 mm. 3The above. HT29 expresses CAPRIN-1 protein on the cell membrane surface, and as shown in Example 4, conjugate 7 and conjugate 10 bind to HT29. Conjugate 7 and conjugate 10 were administered to HT29-bearing mice via the tail vein at 0.3 mg / kg, once a week, for a total of 2 times. In addition, according to the method described in Example 3, a solution containing a conjugate of trastuzumab (Chugai Pharmaceutical Co., Ltd.) and SDG-1910 was prepared in a manner that binds the same degree of drugs as the conjugate using an antibody against CAPRIN-1, and an equal amount was administered to the above-mentioned cancer-bearing mice as a comparative control. HT29 expresses HER2 protein, which is the target antigen of trastuzumab, on the cell membrane surface, and the above-mentioned conjugate of trastuzumab and PBD can specifically bind. PBS(-) was administered to the negative control cancer-bearing mice.

[0326] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor) 2 × 0.5. The evaluation results showed that on the 7th day after the start of administration, when the tumor volume of the negative control was taken as 100%, the mice administered with conjugate 7 and conjugate 10 had a tumor volume of less than 50%. When the tumor volume of cancer-bearing mice administered with only the antibody against CAPRIN-1 used in the above conjugate was taken as 100%, the tumor volume of the cancer-bearing mice administered with the above conjugate was less than 50%. On the other hand, the tumor volume of mice administered with a solution containing a conjugate of trastuzumab and PBD as a comparative control was more than 80% relative to the negative control. Furthermore, when the tumor volume of cancer-bearing mice administered with only trastuzumab was taken as 100%, the tumor volume of cancer-bearing mice administered with the above conjugate was less than 75%.

[0327] The results of this evaluation showed that the conjugate with SDG-1910 prepared in Example 3 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group). Furthermore, it was shown that the conjugate of trastuzumab and SDG-1910 prepared as a comparative control had a significantly stronger antitumor effect.

[0328] (Example 8) Preparation of a conjugate of anti-CAPRIN-1 antibody and a PBD derivative (Tesirine)

[0329] In the same manner as in Example 3, after the lysine residue of the anti-CAPRIN-1 polyclonal antibody #1 described in Example 1 was bound to SATA as a spacer, Tesirine shown in the following chemical structure, which is one of the PBD derivatives, was bound to the antibody via a maleimide group, thereby obtaining a solution containing a conjugate (Conjugate 54). Similarly, a conjugate of the humanized antibody #1 as the anti-CAPRIN-1 monoclonal antibody described in Example 2 and the Tesirine was prepared to obtain a solution containing a conjugate (Conjugate 60). Similarly, conjugates with Tesirine derivatives were prepared for anti-CAPRIN-1 polyclonal antibodies #2 to #6 and humanized antibodies #2 to #47 as anti-CAPRIN-1 monoclonal antibodies. By the same operation as above, solutions containing conjugates with the Tesirine were obtained for the rabbit control antibody and human IgG control antibody described in Example 1 that do not react with the CAPRIN-1 protein (Control Conjugate 3, Control Conjugate 4). The prepared conjugates were evaluated in the same manner as in Examples 4 to 6. As a result, it was confirmed that the conjugates showed specific reactivity to CAPRIN-1 protein and CAPRIN-1-expressing cancer cells, internalized, and had antitumor activity.

[0330]

[0331] (Example 9) Anti-tumor effect of the conjugate of anti-CAPRIN-1 antibody and PBD derivative (Tesirine) in cancer-bearing mice

[0332] The antitumor effects of conjugates 60 and 63 prepared in Example 8 were evaluated in vivo in cancer-bearing mice. The antitumor effects of the conjugates of the present invention were studied using NOD-SCID mice transplanted with human cancer cells expressing CAPRIN-1 protein. 7 Individual colorectal cancer cells HT29 were mixed with Matrigel (SIGMA) and transplanted subcutaneously to allow the tumor to grow to 100-200 mm 3 HT29 expresses CAPRIN-1 protein on the cell membrane surface, and conjugate 60 and conjugate 63 bind to HT29. Conjugate 60 and conjugate 63 were administered once via the tail vein to HT29 cancer-bearing mice at 2 mg / kg and 6 mg / kg, respectively. Negative control cancer-bearing mice were administered PBS (-).

[0333] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor)2 × 0.5. The evaluation results showed that after the 7th day from the start of administration, when the tumor volume of the negative control was taken as 100%, the tumor volume of the mice administered with conjugate 60 and conjugate 632 mg / kg was less than 50% of the tumor volume. On the other hand, in the cancer-bearing mice administered with conjugate 60 and conjugate 636 mg / kg, the tumor regressed.

[0334] The results of this evaluation showed that the conjugate with Tesirine prepared in Example 8 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group).

[0335] (Example 10) Preparation of a conjugate of an anti-CAPRIN-1 antibody and an IGN derivative (DGN549)

[0336] In the same manner as in Example 3, after the lysine residue of the anti-CAPRIN-1 polyclonal antibody #1 described in Example 1 was bound to SATA as a spacer, DGN549, which is one of the IGN derivatives and has the following chemical structure, was bound to the antibody via a maleimide group, thereby obtaining a solution containing a conjugate (conjugate 107). Similarly, a conjugate of the humanized antibody #1 as the anti-CAPRIN-1 monoclonal antibody described in Example 2 and the above-mentioned DGN549 was prepared to obtain a solution containing a conjugate (conjugate 113). Similarly, conjugates of anti-CAPRIN-1 polyclonal antibodies #2 to #6 and humanized antibodies #2 to #47 as anti-CAPRIN-1 monoclonal antibodies and DGN549 were prepared. By the same operation as above, solutions containing conjugates of the rabbit control antibody and human IgG control antibody described in Example 1 that do not react with the CAPRIN-1 protein and DGN549 were obtained in the same manner (control conjugate 5, control conjugate 6). The prepared conjugates were evaluated in the same manner as in Examples 4 to 6. As a result, the specific reactivity and internalization of the conjugates to CAPRIN-1 protein and CAPRIN-1-expressing cancer cells, and the antitumor activity of the conjugates were confirmed.

[0337]

[0338] (Example 11) Anti-tumor effect of the conjugate of anti-CAPRIN-1 antibody and IGN (DGN549) in cancer-bearing mice

[0339] The antitumor effects of conjugates 107 and 113 prepared in Example 10 were evaluated in vivo in cancer-bearing mice. The antitumor effects of the conjugates of the present invention were studied using NOD-SCID mice transplanted with human cancer cells expressing CAPRIN-1 protein. 7 FaDu, a human head and neck cancer cell line, was mixed with Matrigel (SIGMA) and transplanted subcutaneously to allow the tumor to grow to 100-200 mm. 3 The above. FaDu expresses CAPRIN-1 protein on the cell membrane surface, and conjugates 107 and 113 bind to FaDu. Conjugates 107 and 113 were administered once via the tail vein to FaDu cancer-bearing mice at 2 mg / kg and 6 mg / kg, respectively. Negative control cancer-bearing mice were administered PBS(-).

[0340] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor) 2 × 0.5. The evaluation results showed that after the 7th day from the start of administration, when the tumor volume of the negative control was taken as 100%, the tumor volume of the mice administered with conjugate 103 and conjugate 1172 mg / kg was less than 50% of the tumor volume. On the other hand, in the cancer-bearing mice administered with conjugate 103 and conjugate 1176 mg / kg, the tumor regressed.

[0341] The results of this evaluation showed that the conjugate with DGN549 prepared in Example 10 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group).

[0342] (Example 12) Anti-tumor effect of the conjugate of anti-CAPRIN-1 antibody and IGN (DGN549) in cancer-bearing mice

[0343] The antitumor effects of conjugates 107 and 113 prepared in Example 10 were evaluated in vivo in cancer-bearing mice. The antitumor effects of the conjugates of the present invention were studied using CB-17SCID mice transplanted with human cancer cells expressing CAPRIN-1 protein. 6 Personal ovarian cancer cells TOV112D were mixed with Matrigel (SIGMA) and transplanted subcutaneously, and allowed to grow until the tumor was an average of 150-200 mm. 3TOV112D expresses CAPRIN-1 protein on the cell membrane surface, and conjugates 107 and 113 bind to TOV112D. Conjugates 107 and 113 were administered once via the tail vein at 0.5 mg / kg to mice bearing TOV112D cancer. Negative control cancer-bearing mice were administered PBS (-).

[0344] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor) 2 The evaluation results showed that after 15 days from the start of administration, the tumor volume of the tumor-bearing mice administered with conjugate 103 and conjugate 117 was 13% or less, when the tumor volume of the negative control was taken as 100%.

[0345] The results of this evaluation showed that the conjugate with DGN549 prepared in Example 10 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group).

[0346] (Example 13) Anti-tumor effect of the conjugate of anti-CAPRIN-1 antibody and IGN (DGN549) in cancer-bearing mice

[0347] The antitumor effects of conjugates 107 and 113 prepared in Example 10 were evaluated in vivo in cancer-bearing mice. The antitumor effects of the conjugates of the present invention were studied using Balb / c-nu / nu (Balb / c nude) mice transplanted with human cancer cells expressing CAPRIN-1 protein. The following cancer-bearing mice were prepared: 5×10 6 The UMUC3 cells of the individual bladder cancer cells were mixed with Matrigel (SIGMA) and transplanted subcutaneously, and allowed to grow until the tumor was an average of 120 to 150 mm. 3 UMUC3 expresses CAPRIN-1 protein on the cell membrane surface, and conjugates 107 and 113 bind to UMUC3. Conjugates 107 and 113 were administered once via the tail vein at 0.5 mg / kg to UMUC3 cancer-bearing mice. Negative control cancer-bearing mice were administered PBS (-).

[0348] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor) 2The evaluation results showed that after 15 days from the start of administration, the tumor volume of the tumor-bearing mice administered with conjugate 103 and conjugate 117 was 11% or less, when the tumor volume of the negative control was taken as 100%.

[0349] The results of this evaluation showed that the conjugate with DGN549 prepared in Example 10 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group).

[0350] (Example 14) Preparation of conjugates of anti-CAPRIN-1 antibodies and PDD derivatives

[0351] In the same manner as in Example 3, the anti-CAPRIN-1 polyclonal antibody #1 described in Example 1 was bound to the lysine residues with SATA as a spacer according to a conventional method, and then the compound ((S)-N-(4-aminophenyl)-4-(4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine) described in Example 41 of U.S. Patent No. 10975072B2, which is one of the PDDs, was bound to the lysine residues with SATA as a spacer according to a conventional method. A compound having the following chemical structure obtained by binding (1-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2-(4-(2- The prepared conjugates were evaluated in the same manner as in Examples 4 to 6. As a result, the specific reactivity and internalization of the conjugates to CAPRIN-1 protein and CAPRIN-1-expressing cancer cells, and the antitumor activity of the conjugates were confirmed.

[0352]

[0353] (Example 15) Anti-tumor effect of the conjugate of anti-CAPRIN-1 antibody and PDD derivative in cancer-bearing mice

[0354] The antitumor effects of conjugates 160 and 166 prepared in Example 12 were evaluated in vivo in cancer-bearing mice. The antitumor effects of the conjugates of the present invention were studied using NOD-SCID mice transplanted with human cancer cells expressing CAPRIN-1 protein. 7 FaDu, a human head and neck cancer cell line, was mixed with Matrigel (SIGMA) and transplanted subcutaneously to allow the tumor to grow to 100-200 mm. 3 The above. FaDu expresses CAPRIN-1 protein on the cell membrane surface, and conjugates 160 and 166 bind to FaDu. Conjugates 160 and 166 were administered once via the tail vein to FaDu cancer-bearing mice at 6 mg / kg and 12 mg / kg, respectively. Negative control cancer-bearing mice were administered PBS(-).

[0355] The size of the tumor in the tumor-bearing mice after administration was measured over time using a vernier caliper, and the tumor volume was calculated according to a conventional method using the formula: (length of the long axis of the tumor) × (length of the short axis of the tumor) 2 × 0.5. The evaluation results showed that after 30 days from the start of administration, when the tumor volume of the negative control was taken as 100%, the tumor volume of the mice administered with 6 mg / kg of conjugate 160 and 166 was less than 50% of the tumor volume. On the other hand, the tumor volume of the tumor-bearing mice administered with 12 mg / kg of conjugate 160 and conjugate 166 was less than 30%.

[0356] The results of this evaluation showed that the conjugate with a PDD derivative prepared in Example 12 using an antibody against CAPRIN-1 exerted a significantly stronger antitumor effect than the negative control (untreated control group).

Claims

1. Antibodies or fragments thereof and benzodiazepines The antibody or fragment thereof is immunoreactive with a CAPRIN-1 protein having an amino acid sequence represented by any of the even-numbered sequence numbers from SEQ ID NOs. 2 to 30, or an amino acid sequence having a sequence identity of 80% or more to the amino acid sequence.

2. The conjugate according to claim 1, wherein the antibody or fragment thereof is immunoreactive with a partial polypeptide of a CAPRIN-1 protein, wherein the partial polypeptide of the CAPRIN-1 protein has an amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, and 309, or an amino acid sequence having a sequence identity of 80% or more thereto. The conjugate according to claim 1 or 2, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

4. The conjugate according to any one of claims 1 to 3, wherein the antibody or fragment thereof is any one of the following (A) to (M), (A) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 36, 37 and 38 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 40, 41 and 42 as CDR1, CDR2 and CDR3, respectively, (B) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 44, 45 and 46 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 48, 49 and 50 as CDR1, CDR2 and CDR3, respectively, (C) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 52, 53 and 54 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 56, 57 and 58 as CDR1, CDR2 and CDR3, respectively, (D) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 60, 61 and 62 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 64, 65 and 66 as CDR1, CDR2 and CDR3, respectively, (E) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 170, 171 and 172 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 173, 174 and 175 as CDR1, CDR2 and CDR3, respectively, (F) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementarity determining regions of SEQ ID NOs: 176, 177 and 178 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 as CDR1, CDR2 and CDR3, respectively, (G) an antibody or a fragment thereof which is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 182, 183 and 184 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 185, 186 and 187 as CDR1, CDR2 and CDR3, respectively, (H) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 188, 189 and 190 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 191, 192 and 193 as CDR1, CDR2 and CDR3, respectively, (I) an antibody or a fragment thereof comprising a heavy chain variable region and a light chain variable region, which is immunoreactive with a CAPRIN-1 protein, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 146, 147 and 148 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 149, 150 and 151 as CDR1, CDR2 and CDR3, respectively; (J) an antibody or a fragment thereof comprising a heavy chain variable region and a light chain variable region, which is immunoreactive with a CAPRIN-1 protein, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 272, 273 and 274 The complementarity determining regions of the heavy chain variable region and the light chain variable region are as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of sequence numbers 275, 276 and 277 as CDR1, CDR2 and CDR3, respectively. (K) an antibody or a fragment thereof comprising a heavy chain variable region and a light chain variable region and having immunoreactivity with the CAPRIN-1 protein, the heavy chain variable region comprises the complementarity determining regions of sequence numbers 290, 291 and 292 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of sequence numbers 293, 294 and 295 as CDR1, CDR2 and CDR3, respectively. (L) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementary determining regions of SEQ ID NOs: 301, 302, and 303 as CDR1, CDR2, and CDR3, respectively, and the light chain variable region comprises the complementary determining regions of SEQ ID NOs: 305, 306, and 307 as CDR1, CDR2, and CDR3, respectively, (M) An antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the complementarity determining regions of SEQ ID NOs: 134, 135 and 136 as CDR1, CDR2 and CDR3, respectively, and the light chain variable region comprises the complementarity determining regions of SEQ ID NOs: 137, 138 and 139 as CDR1, CDR2 and CDR3, respectively.

5. The conjugate according to any one of claims 1 to 4, wherein the antibody or fragment thereof is any one of the following (a) to (a1), (a) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43, (b) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:47 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:51, (c) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59, (d) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 67, (e) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69, (f) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71, (g) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73, (h) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75, (i) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77, (j) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79, (k) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81, (l) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83, (m) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85, (n) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87, (o) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89, (p) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:91, (q) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:93, (r) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:95, (s) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:97, (t) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:99, (u) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101, (v) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103, (w) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105, (x) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107, (y) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109, (z) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111, (aa) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113, (ab) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115, (ac) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117, (ad) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119, (ae) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121, (af) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123, (ag) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125, (ah) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127, (ai) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129, (aj) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131, (ak) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133, (a1) An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

304. The conjugate according to any one of claims 1 to 5, wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single-chain antibody.

7. The conjugate according to any one of claims 1 to 6, wherein the antibody or fragment thereof is conjugated to a benzodiazepine Classes are connected by joints.

8. The conjugate according to any one of claims 1 to 7, wherein the benzodiazepine Benzodiazepines with antitumor activity kind.

9. The conjugate according to any one of claims 1 to 8, wherein the benzodiazepine Pyrrolobenzodiazepine PBD, indolebenzodiazepine IGN, Pyridobenzodiazepine PDD or isoquinoline benzodiazepine Class (IQB).

10. The conjugate according to any one of claims 1 to 9, wherein the benzodiazepine The classes are DSB-120, SJG-136 (SG2000), DC-81, DSB-120, SJG-136, SG2057, SG2202, SG2285, SGD-1882, SGD-1910, SG3199, SG3249, SG2219, IMGN779, IMGN632, (S)-N-(4-aminophenyl)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazo -3-yl)oxy)butyrylamide)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide, D211, D221, D231, GWL-78, KMR-28-39, or derivatives thereof.

11. The conjugate according to claim 10, wherein the derivative is Tesirine which is a derivative of SG3249, Talirine which is a derivative of SGD-1910, SG3364, SG3227, SG3140, i.e. MC-Phe-Lys-PAB-SG2057, SG3170, SG3203, i.e. MC-Phe-Lys-PAB-SG2057, SG3231, SG3400, SG3376, DGN642, DGN549, FGX5-67 or FGX-2-62, FGX11-38.

12. A pharmaceutical composition for treating and / or preventing cancer, comprising the conjugate according to any one of claims 1 to 11 as an active ingredient. 13 . The pharmaceutical composition according to claim 12 , wherein the cancer is a cancer expressing CAPRIN-1 protein on the cell membrane surface.

14. The pharmaceutical composition of claim 12 or 13, wherein the cancer is breast cancer, kidney cancer, pancreatic cancer, colorectal cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, bile duct cancer, sarcoma, mast cell tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, head and neck cancer, or urothelial carcinoma.

15. A method for treating and / or preventing cancer, comprising administering the conjugate according to any one of claims 1 to 11 or the pharmaceutical composition according to any one of claims 12 to 14 to a subject.

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