Pharmaceutical composition for treating and / or preventing cancer

Abstract

Provided is a conjugate obtained by binding topoisomerase I inhibitor and an antibody or a fragment thereof having immunological reactivity with an amino acid sequence represented by any one of even SEQ ID NOs: 2 to 30 or a CAPRIN -1 protein having an amino acid sequence having 80% or more sequence identity with the antibody. The conjugate can be used as an antibody-drug complex (ADC) for treating and / or preventing cancer.

Description

Pharmaceutical composition for treating and / or preventing cancer

[0001] The present invention relates to a conjugate of an antibody against CAPRIN-1 protein or a fragment thereof with a topoisomerase I inhibitor, and to a pharmaceutical use thereof as a therapeutic and / or preventive agent for cancer, etc.

[0002] Various antibody drugs targeting specific antigen proteins on cancer cells have been applied to cancer treatment as cancer therapeutic agents 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 this CAPRIN-1 protein are known to be promising pharmaceutical applications for the treatment and / or prevention of cancer (Patent Document 1).

[0003] In recent years, studies have been conducted to enhance the efficacy of antibody drugs against cancer. In particular, as one of the means for enhancing the efficacy of antibodies against CAPRIN-1 protein against cancer, a combination therapy of an antibody against CAPRIN-1 protein with topotecan or irinotecan, which are types of topoisomerase I inhibitors, is known (Patent Document 2).

[0004] In addition, the development of antibody-drug conjugates (ADCs), which conjugate antibodies with drugs that have strong killing ability directly against cancer cells, is actively progressing. There are few successful examples of ADCs using topoisomerase I inhibitors, but Trodelvy has developed an ADC in which SN38 is linked to an antibody against TROP-2. ＴＭ(Sacituzumab govitecan-hziy) has been approved for use in patients with metastatic triple-negative breast cancer, and has been shown to reduce the risk of death by approximately half (49%) (Non-Patent Document 1). Furthermore, trastuzumab deruxtecan (T-DXd), an ADC in which deruxtecan, a topoisomerase I inhibitor, is linked to an antibody against HER2, has been approved for use in patients with HER2-positive, inoperable or recurrent breast cancer who have received chemotherapy, inoperable or recurrent breast cancer with low HER2 expression who have received chemotherapy, inoperable, advanced or recurrent HER2-mutation-positive non-small cell lung cancer that has progressed after chemotherapy, and inoperable, advanced or recurrent HER2-positive gastric cancer that has progressed after chemotherapy. In patients with HER2-positive, inoperable or recurrent breast cancer who have received treatment with trastuzumab and a taxane anti-cancer drug, progression-free survival was significantly improved compared to trastuzumab emtansine (Trastuzumab emtansine), an ADC in which trastuzumab is linked to emtansine (DM1). A significant prolongation was observed compared to T-DM1 (emtansine, T-DM1) (Non-patent Document 2).

[0005] WO2010 / 016526 WO2022 / 270524

[0006] GILEAD Press Releases, April 07, 2021 THE LANCET Vol. 401, May 27, 2023

[0007] As mentioned above, ADCs using topoisomerase I inhibitors have already been put to practical use with the aim of enhancing the efficacy of antibody drugs against cancer, but success has been limited to targeting only a few cancers, and the antitumor effect has also been limited. Furthermore, as shown in the Examples of the present specification, the present inventors have newly discovered that the efficacy of a conjugate of trastuzumab and a topoisomerase I inhibitor, which is commercially available as a cancer antibody drug, is limited in enhancing the antitumor effect compared to trastuzumab alone, and that ADC technology using a topoisomerase I inhibitor is not a technology that significantly enhances the antitumor effect of all cancer antibody drugs.

[0008] Therefore, the present invention aims to discover a cancer antibody drug whose efficacy can be significantly enhanced by applying ADC technology that utilizes a topoisomerase I inhibitor, and to create a new ADC that utilizes a topoisomerase I inhibitor and has excellent antitumor effects.

[0009] As a result of intensive research, the present inventors have discovered that a conjugate of an antibody against CAPRIN-1 protein or a fragment thereof with a topoisomerase I inhibitor exhibits an extremely strong antitumor effect compared to an antibody against CAPRIN-1 protein or a fragment thereof alone, and further that the antitumor effect enhancement effect when an antibody against CAPRIN-1 protein or a fragment thereof is conjugated with a topoisomerase I inhibitor is significantly superior to the antitumor effect enhancement effect when an existing cancer antibody pharmaceutical is conjugated with a topoisomerase I inhibitor, thereby completing the present invention.

[0010] Specifically, the present invention has the following features (1) to (12).

[0011] (1) A conjugate comprising an antibody or a fragment thereof immunologically reactive with a CAPRIN-1 protein having an amino acid sequence represented by any of the even-numbered SEQ ID NOs: 2 to 30, or an amino acid sequence having 80% or more sequence identity with said amino acid sequence, and a topoisomerase I inhibitor bound thereto.

[0012] (2) The conjugate according to (1), wherein the antibody or a fragment thereof is immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having 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 80% or more sequence identity to said amino acid sequence.

[0013] (3) The conjugate according to (1) or (2), wherein the antibody is a monoclonal antibody or a polyclonal antibody.

[0014] (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): (A) an antibody or fragment comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 36, 37, and 38 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 40, 41, and 42 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein; (B) an antibody or fragment comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 44, 45, and 46 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 48, 49, and 50 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (C) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 52, 53, and 54 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 56, 57, and 58 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with the CAPRIN-1 protein. (D) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 60, 61, and 62 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 64, 65, and 66 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with the CAPRIN-1 protein. (E) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (F) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein.(G) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (H) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (I) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 146, 147, and 148 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 149, 150, and 151 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (J) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 272, 273, and 274 (CDR1, CDR2, and CDR3, respectively), and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 275, 276, and 277 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (K) 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: 290, 291 and 292 and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 293, 294 and 295, and having immunological reactivity with the CAPRIN-1 protein. (L) 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: 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, and having immunological reactivity with the CAPRIN-1 protein.(M) An antibody or fragment thereof comprising a heavy chain variable region containing the complementarity determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 134, 135, and 136, respectively, and a light chain variable region containing the complementarity determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 137, 138, and 139, respectively, and having immunological reactivity with a CAPRIN-1 protein.

[0015] (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): (a) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 43; (b) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 47 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 51; (c) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; (d) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 67; (e) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 69; (f) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 71. (g) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 73; (h) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 75; (i) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 77; (j) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 79; (k) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 81; (l) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 83. (m) an antibody or 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 fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 87; (o) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 89; (p) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 91; (q) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 92 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 93; (r) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 94 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 95; (s) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 96 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 97. (t) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 99; (u) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 101; (v) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 103; (w) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 105; (x) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 107; (y) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 109. (z) an antibody or 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 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 fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 115; (ac) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 117; (ad) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 119; (ae) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 121; (af) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 123; (ag) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 125. (ah) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 127; (ai) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 129; (aj) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 131; (ak) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 133; (al) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 304.

[0016] (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.

[0017] (7) The conjugate according to any one of (1) to (6), wherein the antibody or fragment thereof and the topoisomerase I inhibitor are bound via a linker.

[0018] (8) The conjugate according to any one of (1) to (7), wherein the topoisomerase I inhibitor is exatecan, SN38, or a derivative thereof.

[0019] (9) A pharmaceutical composition for treating and / or preventing cancer, comprising the conjugate according to any one of (1) to (8) as an active ingredient.

[0020] (10) The pharmaceutical composition according to (9), wherein the cancer is a cancer that expresses CAPRIN-1 protein on the cell membrane surface.

[0021] (11) The pharmaceutical composition according to (9) or (10), wherein the cancer is breast cancer, kidney cancer, pancreatic cancer, colon 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 cancer.

[0022] (12) A method for treating and / or preventing cancer, comprising administering to a subject the conjugate according to any one of (1) to (8) or the pharmaceutical composition according to any one of (9) to (11).

[0023] The conjugate of the present invention not only exhibits an extremely strong antitumor effect compared to an antibody against CAPRIN-1 protein or a fragment thereof alone, but also has a significantly superior antitumor effect compared to previously known conjugates of cancer antibody drugs and topoisomerase I inhibitors. Furthermore, the enhancement of the antitumor effect achieved by conjugating an antibody against CAPRIN-1 protein or a fragment thereof with a topoisomerase I inhibitor is significantly superior to the enhancement of the antitumor effect achieved by conjugating an existing cancer antibody drug with a topoisomerase I inhibitor. Therefore, the conjugate of the present invention is effective in treating and preventing cancer.

[0024] The antitumor activity of a conjugate of an antibody against the CAPRIN-1 protein or a fragment thereof (hereinafter referred to as an "anti-CAPRIN-1 antibody") used in the present invention and a topoisomerase I inhibitor can be evaluated by examining the inhibition of tumor growth in vitro or in vivo (specifically, in tumor-bearing animals) as described below.

[0025] In the present invention, the term "conjugate" refers to an antibody and a drug linked by a covalent bond. The antibody and drug may be linked via a linker.

[0026] The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal or polyclonal antibody, preferably a monoclonal antibody. The conjugate of the present invention may be any type of antibody, including a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, or a non-human animal antibody, as long as it can exert antitumor activity.

[0027] The topoisomerase I inhibitor used in the present invention is known as a substance that is toxic to cancer cells by recognizing a specific DNA sequence, binding to the minor groove (DNA minor groove), and inhibiting DNA synthesis and cell proliferation.

[0028] Furthermore, subjects for cancer treatment and / or prevention in the present invention include mammals such as humans, pet animals, livestock, and sport animals, with humans being the preferred subject.

[0029] The following describes the anti-CAPRIN-1 antibody, the topoisomerase I inhibitor, the conjugate of the anti-CAPRIN-1 antibody and the topoisomerase I inhibitor, the pharmaceutical composition using the conjugate, and the method for treating and / or preventing cancer according to the present invention.

[0030] <Anti-CAPRIN-1 Antibody> The anti-CAPRIN-1 antibody used in the present invention refers to an antibody or a fragment thereof that is immunologically reactive with the full-length CAPRIN-1 protein or a fragment thereof. Here, "immunological reactivity" refers to the property of the antibody specifically binding to the CAPRIN-1 protein or a partial polypeptide thereof in vivo.

[0031] Among CAPRIN-1 proteins having an amino acid sequence represented by any of the even-numbered SEQ ID NOS: 2 to 30, which are immunologically reactive 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 the amino acid sequences of canine CAPRIN-1 protein, the amino acid sequences represented by SEQ ID NOS: 2 and 4 are the amino acid sequences of human CAPRIN-1 protein, the amino acid sequence represented by SEQ ID NOS: 16 is the amino acid sequence of bovine CAPRIN-1 protein, the amino acid sequence represented by SEQ ID NOS: 18 is the amino acid sequence of equine CAPRIN-1 protein, the amino acid sequences represented by SEQ ID NOS: 20 to 28 are the amino acid sequences of mouse CAPRIN-1 protein, and the amino acid sequence represented by SEQ ID NOS: 30 is the amino acid sequence of chicken CAPRIN-1 protein.

[0032] Furthermore, the anti-CAPRIN-1 antibody used in the present invention may be immunologically reactive with a variant of the CAPRIN-1 protein having 80% or more, preferably 90% or more, more preferably 95% or more, and even more preferably 99% or more sequence identity with the amino acid sequence represented by any of the even-numbered SEQ ID NOs: 2 to 30. As used herein, "% sequence identity" refers to the percentage (%) of identical amino acids (or bases) relative to the total number of amino acids (or bases) when the two sequences are aligned to maximize similarity, with or without introducing gaps.

[0033] The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.

[0034] Polyclonal antibodies immunologically reactive with the full-length CAPRIN-1 protein or a fragment thereof (anti-CAPRIN-1 polyclonal antibodies) can be obtained, for example, by immunizing mice, human antibody-producing mice, rats, rabbits, chickens, etc. with natural CAPRIN-1 protein, a fusion protein with GST or the like, or a partial peptide thereof, and then purifying the resulting serum using ammonium sulfate precipitation, protein A, protein G, a DEAE ion exchange column, an affinity column to which the CAPRIN-1 protein or a partial peptide is bound, or the like.

[0035] The full-length CAPRIN-1 protein or a fragment thereof used in the immunization can be obtained by accessing the nucleotide sequence and amino acid sequence of CAPRIN-1 and its homologs, for example, 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). Methods 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.

[0036] A monoclonal antibody immunologically reactive with the full-length CAPRIN-1 protein or a fragment thereof (anti-CAPRIN-1 monoclonal antibody) can be obtained, for example, by immunizing a mouse with SK-BR-3 breast cancer cells expressing the CAPRIN-1 protein or the full-length CAPRIN-1 protein or a fragment thereof, fusing spleen cells isolated from the mouse with myeloma cells, and selecting a clone producing an anti-CAPRIN-1 monoclonal antibody from the resulting fused cells (hybridoma). The antibody produced by the selected hybridoma can be obtained by a method similar to the method for purifying polyclonal antibodies described above.

[0037] The antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies, and non-human animal antibodies.

[0038] Human antibodies can be obtained by sensitizing human lymphocytes infected with EB virus with a protein, protein-expressing cells, or a lysate thereof, fusing the sensitized lymphocytes with myeloma cells such as human-derived U266 cells, and then obtaining antibodies immunologically reactive with the full-length CAPRIN-1 protein or a fragment thereof from the resulting fused cells.

[0039] A humanized antibody is a modified antibody, also known as a reshaped human antibody. Humanized antibodies are constructed by grafting the complementarity-determining regions (CDRs) of an antibody derived from an immunized animal onto the CDRs of a human antibody. Genetic recombination, a common technique for this purpose, is well known. Specifically, for example, a DNA sequence designed to link the CDRs of a mouse or rabbit antibody with the framework regions of a human antibody is synthesized by PCR from several oligonucleotides engineered to have overlapping ends. The resulting DNA is ligated to DNA encoding the constant regions of a human antibody, incorporated into an expression vector, and then introduced into a host for production (see EP 239400 and WO 96 / 02576). The framework regions of the human antibody linked via the CDRs are selected so that the CDRs form a good antigen-binding site. If necessary, amino acids in the framework regions of the variable regions of the antibody may be substituted so that the complementarity-determining regions of the reshaped human antibody form an appropriate antigen-binding site (Sato K. et al., Cancer Research 1993, 53:851-856). Alternatively, they may be substituted with framework regions derived from various human antibodies (see WO99 / 51743).

[0040] Antibodies are typically heteromeric glycoproteins containing at least two heavy chains and two light chains. Antibodies consist of two identical light chains and two identical heavy chains. Heavy chains have a heavy chain variable region at one end, followed by several constant regions. Light chains have a light chain variable region at one end, followed by several constant regions. The variable regions exhibit specific variable regions called complementarity-determining regions (CDRs) that confer binding specificity to the antibody. Portions of the variable regions that are relatively conserved are called framework regions (FRs). Complete heavy and light chain variable regions each contain four FRs connected by three CDRs (CDR1 to CDR3).

[0041] The sequences of the constant and variable regions of human-derived heavy and light chains are available from NCBI (USA: GenBank, UniGene, etc.). For example, reference can be made to the sequences of the human IgG1 heavy chain constant region under accession number J00228, the human IgG2 heavy chain constant region under accession number J00230, the human light chain κ constant region under accession numbers V00557, X64135, X64133, etc., and the human light chain λ constant region under accession numbers X64132, X64134, etc.

[0042] A chimeric antibody is an antibody produced by combining sequences derived from different animals, such as an antibody consisting of the heavy chain variable region and light chain variable region of a mouse antibody and the heavy chain variable region and light chain variable region constant region of a human antibody. Chimeric antibodies can be produced using known methods, for example, by linking DNA encoding an antibody V region with DNA encoding a human antibody C region, incorporating the resultant into an expression vector, and introducing the vector into a host for production.

[0043] Non-human animal antibodies can be obtained by immunizing an animal with a sensitizing antigen according to known methods. A typical method is to inject the sensitizing antigen intraperitoneally, intradermally, or subcutaneously into an animal such as a mouse. When injecting the sensitizing antigen, the antigen is mixed with an appropriate amount of various adjuvants, such as Freund's complete adjuvant (CFA), and administered to the animal multiple times. After immunizing an animal and confirming that the serum contains anti-CAPRIN-1 antibodies, the serum can be obtained and purified, as described above, by ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange columns, affinity columns coupled with CAPRIN-1 protein or partial peptides, or the like. Furthermore, monoclonal antibodies can be obtained from non-human animals by collecting immune cells from the immunized animal and fusing them with myeloma cells. The fusion of the immune cells with myeloma cells can be carried out according to known methods (see Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).

[0044] The antibodies used in the present invention can also be obtained as recombinant antibodies produced by cloning an antibody gene from a hybridoma, incorporating it into a suitable vector, and introducing it into a host using genetic engineering techniques (see Carl, A.K., Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990).

[0045] The anti-CAPRIN-1 antibody used to obtain the conjugate of the present invention may have amino acids in the variable region (e.g., FR) or constant region substituted with other amino acids. The amino acid substitutions are one or more, for example, fewer than 15, fewer than 10, 8 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer amino acids, preferably 1 to 9 amino acids. The substituted antibody should have the same or higher antigen-specific binding properties and antigen-binding affinity as the unsubstituted antibody, and should not cause rejection reactions when administered to humans.

[0046] The anti-CAPRIN-1 antibody used in the present invention is expected to have a stronger antitumor effect if it has a higher binding affinity with the CAPRIN-1 protein on the surface of cancer cells. 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 It is desirable that:

[0047] The binding ability of the anti-CAPRIN-1 antibody used in the present invention to effector cells can be improved by substituting one, two, or several amino acids in the heavy chain constant region of the antibody, or by removing fucose bound to N-acetylglucosamine in the N-glycoside-linked sugar chain bound to the heavy chain constant region. The above may be achieved by amino acid substitution alone, or may be a composition with an antibody bound to fucose.

[0048] Antibodies in which one, two, or several amino acids in the heavy chain constant region have been substituted can be produced by referring to, for example, WO2004 / 063351, WO2011 / 120135, U.S. Patent No. 8,388,955, WO2011 / 005481, U.S. Patent No. 6,737,056, and WO2005 / 063351.

[0049] An antibody from which fucose bound to N-acetylglucosamine in the N-glycoside-linked sugar chain in the heavy chain constant region has been removed, or a cell producing such an antibody, 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. An antibody from which fucose bound to N-acetylglucosamine in the N-glycoside-linked sugar chain bound to the heavy chain constant region has been removed, or a composition of an antibody to which fucose has been bound, or a cell producing such an antibody, can be prepared with reference to, for example, U.S. Patent No. 8,642,292.

[0050] The anti-CAPRIN-1 polyclonal antibody, anti-CAPRIN-1 monoclonal antibody, antibody production method, purification method, and method for producing the CAPRIN-1 protein or its partial polypeptide used in immunization used in the present invention are described in WO2010 / 016526, WO2011 / 096517, W2011 / 096528, W O2011 / 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.

[0051] Specific examples of the anti-CAPRIN-1 antibody of the present invention include those described in the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, and WO2013 / 01889 1, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176, and WO2015 / 020212, but preferred anti-CAPRIN-1 antibodies include the following:

[0052] An antibody or fragment thereof that is immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4, or an amino acid sequence that has 80% or more (preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and still more preferably 99% or more) sequence identity with said amino acid sequence.

[0053] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence represented by SEQ ID NO: 31 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. An antibody or fragment thereof immunologically reactive with a CAPRIN-1 protein, preferably comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 36, 37, and 38 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 40, 41, and 42 (CDR1, CDR2, and CDR3, respectively), or an antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 140, 141, and 142 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 143, 144, and 145. or an antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 164, 165, and 166 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 167, 168, and 169 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof has 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 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 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.

[0054] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 33 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 60, 61, and 62 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 64, 65, and 66 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 63 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 67.

[0055] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 32 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 52, 53, and 54 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 56, 57, and 58 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 55 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 59.

[0056] An antibody or fragment thereof that is immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence represented by SEQ ID NO: 34 or an amino acid sequence that has 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said 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 immunologically reactive with the CAPRIN-1 protein; or an antibody or fragment thereof comprises 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 immunologically reactive with the CAPRIN-1 protein. More preferably, the antibody or fragment thereof has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 80 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 81, or an antibody or fragment thereof has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 83.

[0057] An antibody or fragment thereof that is immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence that has 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said 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 immunologically reactive with the CAPRIN-1 protein; or an antibody or fragment thereof comprises 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 immunologically reactive with the CAPRIN-1 protein. More preferably, the antibody or fragment thereof has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 84 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 85, or an antibody or fragment thereof has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 86 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 87.

[0058] An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 44, 45, and 46, and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 48, 49, and 50, and having immunological reactivity with a CAPRIN-1 protein. Preferably, the antibody or fragment thereof comprises the heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 47, and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 51.

[0059] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 296 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 146, 147, and 148 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 149, 150, and 151 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 72 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 73.

[0060] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 297 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 272, 273, and 274 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 275, 276, and 277 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 114 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 115.

[0061] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 298 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 290, 291, and 292 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 293, 294, and 295 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 120 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 121.

[0062] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 299 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 301, 302, and 303 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 305, 306, and 307 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 300 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 304.

[0063] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 308 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 134, 135, and 136 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 137, 138, and 139 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 68 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 69.

[0064] An antibody or fragment thereof immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having the amino acid sequence set forth in SEQ ID NO: 309 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) sequence identity with said amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 134, 135, and 136 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity-determining regions of SEQ ID NOs: 137, 138, and 139 (CDR1, CDR2, and CDR3, respectively), and is immunologically reactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 68 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 69.

[0065] In addition, the following anti-CAPRIN-1 antibodies are also preferably used.

[0066] An antibody or 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.

[0067] An antibody or 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.

[0068] An antibody or 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.

[0069] An antibody or 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.

[0070] An antibody or 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.

[0071] An antibody or 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.

[0072] An antibody or 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.

[0073] An antibody or 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.

[0074] An antibody or 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.

[0075] An antibody or 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.

[0076] An antibody or 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.

[0077] An antibody or 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.

[0078] An antibody or 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.

[0079] An antibody or 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.

[0080] An antibody or 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.

[0081] An antibody or 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.

[0082] An antibody or 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.

[0083] An antibody or 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.

[0084] An antibody or 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.

[0085] An antibody or 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.

[0086] An antibody or 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.

[0087] An antibody or 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.

[0088] An antibody or 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.

[0089] An antibody or 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.

[0090] An antibody or 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.

[0091] An antibody or 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.

[0092] An antibody or 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.

[0093] An antibody or 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.

[0094] An antibody or 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.

[0095] An antibody or 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.

[0096] An antibody or 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.

[0097] An antibody or 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.

[0098] An antibody or 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.

[0099] An antibody or 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.

[0100] In the Examples described below, conjugates with topoisomerase I inhibitors were prepared using the above-mentioned polyclonal and monoclonal antibodies against the full-length CAPRIN-1 protein and a portion of the polypeptide region expressed on the cell membrane surface of cancer cells, and their strong antitumor effects were confirmed.

[0101] <Topoisomerase I inhibitors> Topoisomerase I inhibitors reversibly cleave one strand of the DNA double helix, interfering with the action of topoisomerase I, which is involved in DNA replication, DNA folding and unwinding, and DNA unwinding during transcription, and can induce cell death in cancer cells.

[0102] Specific examples of topoisomerase I inhibitors used in the present invention include quinoline alkaloids, and representative compounds include camptothecin and its derivatives. Specific examples of camptothecin derivatives include those that retain the D-ring pyrrolidone, which is the active center of camptothecin.

[0103] Specific examples of topoisomerase I inhibitors include exatecan (DX-8951), exatecan derivatives (execatecan methanesulfonic acid dihydrate (DX-8951f), MAAAA-1181a (DXd payload), deruxtecan, Dxd-d5), SN38 (7-ethyl-10-hydroxycamptothecin), SN38 derivatives (SN38 glucuronide, SN38-COOH, SN38 glucuronide-13C6, SN38-d3, SN38-CM2, SN38-CO-DMEDA TFA, MAC glucuronide phenol-linked SN38, MAC glucuronide α-hydroxy lactone-linked SN38, PH-HG-005-5, Antitumor agent-F10, TP3011 (CH0793011)), camptothecin, 10-hydroxycamptothecin, 7-n-butyl-10-aminocamptothecin, 7-n-butyl-9-amino-10,11-methylenedioxycamptothecin, T-2513, T-0128, irinotecan (CPT-11), topotecan (also called nogitecan).), Roototecan, ST1481, CKD602, DB67, BNP135, Antitumor agent-102, PNU-159682, 9-aminocamptothecin, amsacrine, belotecan, BN-80915, diflomotecan, edotecarin, epirubicin, gimatecan, aclacinomycin A hydrochloride, becatecarin, β-lapachone, camptothecin-20(S)-O-propionic acid, CH-0793076 (TP3076) hexacyclic camptothecin analog, coralline chloride, DRF-1042, gimatecan, indimitecan, LMP-400, intoplicin (RP 60475), karenitecin, LMP744 and its hydrochloride, Luotonin A, Luotonin F, Mauritianin, Namitecan (ST-1968), Podocarpus flavone A, Proscillaridin A, Pyrazoloacridine (NSC 366140), Rebeccamycin, Rubitecan, SW044248, TAS-103 (BMS-247615), TAS-103 dihydrochloride (BMS-247615 dihydrochloride), Topoisomerase I inhibitor 8 (CAS No. 210346-40-0), camptothecin derivatives (NSC100880, NSC603071, NSC107124, NSC643833, NSC629971, NSC295500, NSC249910, NSC606985, NSC74028, NSC176323, NSC295501, NSC606172, NSC606173, NSC610458, NSC618939, NSC610457, NSC610459, NSC606499, NSC610456 , NSC364830 and NSC606497), morpholine isoxorubicin, (7S)-14-(3-aminobicyclo[1.1.1]pentan-1-yl)-7-ethyl-7-hydroxy-2H,10H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H,13H)-dione, CT-2106, DX-310, PNU-159682, AZ14170132 or derivatives thereof.

[0104] The topoisomerase I inhibitor used in the present invention may be a conjugated compound with a linker. Specific examples of the topoisomerase I inhibitor and the conjugated compound include camptothecin analogues, and are described in WO 1993 / 018045, WO 2000 / 046228, WO 2002 / 083682, WO 2005 / 040170, WO 2005 / 110423, WO 2005 / 085251, WO 2006 / 135687, WO 2010 / 043880, WO 2013 / 055990, WO 2013 / 055993, WO 2014 / 057072, WO 2014 / 159981, WO 2014 / 0571 No. 20, WO2015 / 052532, WO2015 / 166289, WO2015 / 181559, WO2017 / 020 No. 972, WO2017 / 059289, WO2017 / 137555, WO2017 / 137556, WO2017 / 18 No. 6894, WO2018 / 031662, WO2018 / 069490, WO2018 / 091646, WO2018 / 1 No. 46188, WO2018 / 146189, WO2018 / 18234, WO2018 / 192944, WO2019 / 0 No. 34764, WO2019 / 065964, WO2019 / 096788, WO2019 / 104289, WO2019 / No. 126691, WO2019 / 224340, WO2020 / 006722, WO2020 / 006722, WO2020 / 079229, WO2020 / 079239, WO2020 / 100954, WO2020 / 141923, WO202 No. 0 / 152462, WO2020 / 1964474, WO2020 / 196712, WO2021 / 137646, WO20 No. 22 / 063853, WO2021 / 188896, WO2021 / 067403, WO2018 / 217227, WO2 No. 018 / 187074, WO2018 / 183041, WO2018 / 156634, WO2018 / 102212, WO 2018 / 031408, WO2017 / 189279, WO2021 / 173773, WO2017 / 139623, W O2017 / 034906, WO2017 / 004144, WO2016 / 210108, WO2016 / 201300,WO2016 / 172427, WO2016 / 111751, WO2016 / 077061, WO2016 / 057398, WO2016 / 003869, WO2015 / 130416, WO2015 / 069430, WO2015 / 047510, WO2 015 / 012904、WO2014 / 165506、WO2014 / 127200、WO2014 / 124227、WO2014 / 092804、WO2013 / 085893、WO2013 / 043800、WO2013 / 012894、WO2012 / 024223, WO2011 / 123428, WO2011 / 072124, WO2011 / 068845, WO2011 / 034660, WO2010 / 117984, WO2010 / 093395, WO2010 / 017500, WO2009 / 1001 94, WO2005 / 086612, WO2005 / 077071, WO2005 / 069994, WO2005 / 021494, WO2005 / 014618, WO2004 / 110390, WO2004 / 074434, WO2004 / 054622, WO2023 / 088235, WO2022 / 236136, WO2021 / 102092, WO2021 / 005583, WO2020 / 219287, WO2017 / 156183, WO2015 / 048365, WO2015 / 000240, WO2 014 / 210082、WO2014 / 067960、WO2014 / 064654、WO2013 / 067449、WO2012 / 139487、WO2012 / 136144、WO2012 / 096832、WO2012 / 007619、WO2011 / 154574, WO2010 / 148138, WO2010 / 060098, WO2009 / 098116, WO2008 / 038944, WO2008 / 012003, WO2008 / 011994, WO2008 / 011992, WO2007 / 1042 14, WO2007 / 085370, WO2007 / 050892, WO2007 / 048002, WO2007 / 015259, WO2006 / 092230, WO2006 / 082053, WO2005 / 117881, WO2005 / 117879,WO2005 / 110487, WO2005 / 070936, WO2005 / 061517, WO2005 / 044821, WO2004 / 092205, WO2004 / 083214, WO2004 / 073601, WO2004 / 056398, WO2 003 / 101998、WO2003 / 101406、WO2003 / 097356、WO2003 / 033525、WO2003 / 031467、WO2002 / 070525、WO2002 / 056885、WO2002 / 022618、WO2001 / 008663, WO2000 / 073305, WO2000 / 066125, WO2000 / 061187, WO2000 / 053607, WO2000 / 007605, WO1999 / 024430, WO1999 / 017805, WO1999 / 0025 30, WO1998 / 035940, WO1998 / 014468, WO1998 / 007727, WO1997 / 031003, WO1997 / 028165, WO1997 / 028164, WO1997 / 016454, WO1996 / 039143, WO1996 / 037496, WO1996 / 031513, WO1996 / 021666, WO1996 / 011005, WO1995 / 028404, WO1995 / 022549, WO1995 / 010304, WO1994 / 025466, WO1 994 / 011377, WO2023 / 175483, WO2023 / 042097, WO2022 / 054009, WO2020 / 240467, WO2019 / 219891, WO2023 / 126823, WO2023 / 126822, WO2023 / 089527, WO2022 / 074617, WO2022 / 019259, WO2021 / 260583, WO2021 / 260582, WO2021 / 260580, WO2021 / 260579, WO2021 / 260578, WO2019 / 2122 53, WO2013 / 191512, WO2011 / 108909, WO2001 / 049691, WO2010 / 080478, WO2022 / 232834, WO2022 / 094244, WO2021 / 207581, WO2020 / 223536,WO2020 / 186199, WO2020 / 077217, WO2015 / 031541, WO2015 / 031693, WO2015 / 031698, WO2017 / 201204, WO2016 / 070089, WO2017 / 214335, WO2 019 / 236954、WO2022 / 056696、WO2019 / 044946、WO2009 / 089502、WO2009 / 089537、WO2009 / 093242、WO2009 / 097325、WO2009 / 097397、WO2009 / 098469, WO2009 / 099959, WO2009 / 101614, WO2009 / 102433, WO2009 / 108384, WO2009 / 108932, WO2009 / 111950, WO2009 / 121564, WO2009 / 1237 68, WO2009 / 124064, WO2009 / 126310, WO2009 / 126805, WO2009 / 126934, WO2009 / 126950, WO2009 / 135019, WO2009 / 137378, WO2009 / 141050, WO2009 / 141240, WO2009 / 142328, WO2009 / 142810, WO2009 / 145282, WO2009 / 148698, WO2009 / 150248, WO2009 / 152247, WO2009 / 158381, WO2 010 / 005723、WO2010 / 009075、WO2010 / 009139、WO2010 / 009155、WO2010 / 009166、WO2010 / 009335、WO2010 / 014611、WO2010 / 015688、WO2010 / 017035, WO2010 / 019233, WO2010 / 020676, WO2010 / 021750, WO2010 / 021934, WO2010 / 021973, WO2010 / 025337, WO2010 / 033692, WO2010 / 0337 71, WO2010 / 043050, WO2010 / 046900, WO2010 / 048018, WO2010 / 048603, WO2010 / 054398, WO2010 / 057203, WO2010 / 059943, WO2010 / 063630,WO2010 / 063696, WO2010 / 065329, WO2010 / 065893, WO2010 / 067886, WO2010 / 067888, WO2010 / 068720, WO2010 / 068759, WO2010 / 068920, WO2 010 / 071837、WO2010 / 083199、WO2010 / 083220、WO2010 / 083465、WO2010 / 084157、WO2010 / 088650、WO2010 / 088866、WO2010 / 093789、WO2010 / 096175、WO2010 / 098367、WO2010 / 101734、WO2010 / 104595、WO2010 / 105160、WO2010 / 115377、WO2010 / 118200、WO2010 / 121455、WO2010 / 121455 40, WO2010 / 124009, WO2010 / 130224, WO2010 / 131675, WO2010 / 138661, WO2010 / 138686, WO2010 / 138719, WO2010 / 142124, WO2010 / 144371, WO2010 / 144378, WO2010 / 148163, WO2010 / 148288, WO2010 / 149326, WO2010 / 150255, WO2010 / 151653, WO2011 / 000217, WO2011 / 002852, WO2 011 / 009539、WO2011 / 011474、WO2011 / 017998、WO2011 / 023367、WO2011 / 031865、WO2011 / 034954、WO2011 / 035261、WO2011 / 035264、WO2011 / 053750, WO2011 / 054001, WO2011 / 057022, WO2011 / 057064, WO2011 / 062420, WO2011 / 063156, WO2011 / 064152, WO2011 / 066418, WO2011 / 0721 27, WO2011 / 072398, WO2011 / 075185, WO2011 / 076196, WO2011 / 076547, WO2011 / 076880, WO2011 / 079279, WO2011 / 090492, WO2011 / 092708,WO2011 / 094671, WO2011 / 106248, WO2011 / 106702, WO2011 / 109237, WO2011 / 113879, WO2011 / 116226, WO2011 / 117851, WO2011 / 120099, WO2011 / 123524, WO2011 / 124718, WO2011 / 130674, WO2011 / 131314, WO2011 / 131636, WO2011 / 133925, WO2011 / 137516, WO2011 / 140334, WO2011 / 143201, WO2011 / 143399, WO2011 / 143419, WO2011 / 143422, WO2011 / 143423, WO2011 / 143425, WO2011 / 143426, WO2011 / 144742, WO2011 / 1447 56, WO2011 / 146638, WO2011 / 147296, WO2011 / 147409, WO2011 / 150283, WO2011 / 151824, WO2011 / 155501, WO2011 / 163527, WO2012 / 004008, WO2012 / 011693, WO2012 / 011917, WO2012 / 026982, WO2012 / 027204, WO2012 / 028101, WO2012 / 028679, WO2012 / 031457, WO2012 / 031773, WO2012 / 033601, WO2012 / 040623, WO2012 / 060544, WO2012 / 064806, WO2012 / 071554, WO2012 / 075408, WO2012 / 082483, WO2012 / 083594, WO2012 / 094620, WO2012 / 106509, WO2012 / 113571, WO2 012 / 116277、WO2012 / 118978、WO2012 / 121953、WO2012 / 125232、WO2012 / 134446、WO2012 / 135729、WO2012 / 135831、WO2012 / 138938、WO2012 / 140252、WO2012 / 142141、WO2012 / 142926、WO2012 / 145981、WO2012 / 146681、WO2012 / 148799、WO2012 / 149186, WO2012 / 155019, WO2012 / 156310, WO2012 / 158622, WO2012 / 162307, WO2012 / 167099, WO2012 / 167309, WO2012 / 170470, WO2 012 / 170513、WO2012 / 170640、WO2012 / 176145、WO2012 / 178123、WO2012 / 178124、WO2012 / 178125、WO2013 / 000269、WO2013 / 002969、WO2013 / 013404, WO2013 / 016696, WO2013 / 023043, WO2013 / 028543, WO2013 / 028989, WO2013 / 043529, WO2013 / 049719, WO2013 / 049720, WO2013 / 0497 22, WO2013 / 055771, WO2013 / 055913, WO2013 / 056132, WO2013 / 067162, WO2013 / 067881, WO2013 / 067882, WO2013 / 070688, WO2013 / 071053, WO2013 / 071085, WO2013 / 071088, WO2013 / 071090, WO2013 / 071093, WO2013 / 071094, WO2013 / 078392, WO2013 / 089394, WO2013 / 093931, WO2 013 / 094581、WO2013 / 096388、WO2013 / 103703、WO2013 / 106643、WO2013 / 109675、WO2013 / 120856、WO2013 / 122112、WO2013 / 122545、WO2013 / 122751, WO2013 / 124068, WO2013 / 125891, WO2013 / 130683, WO2013 / 131927, WO2013 / 138371, WO2013 / 142427, WO2013 / 142817, WO2013 / 1485 68, WO2013 / 149219, WO2013 / 149538, WO2013 / 151584, WO2013 / 151638, WO2013 / 152227, WO2013 / 152298, WO2013 / 154707, WO2013 / 154774,WO2013 / 157021, WO2013 / 157022, WO2013 / 166366, WO2013 / 167743, WO2013 / 169337, WO2013 / 169479, WO2013 / 170147, WO2013 / 170159, WO2 013 / 170272、WO2013 / 172999、WO2013 / 179144、WO2013 / 188763、WO2013 / 189237、WO2013 / 189266、WO2014 / 004376、WO2014 / 008875、WO2014 / 009872, WO2014 / 012479, WO2014 / 022357, WO2014 / 033136, WO2014 / 036037, WO2014 / 036566, WO2014 / 039787, WO2014 / 041517, WO2014 / 0424 50, WO2014 / 054026, WO2014 / 055493, WO2014 / 055913, WO2014 / 059028, WO2014 / 062604, WO2014 / 062856, WO2014 / 065513, WO2014 / 066002, WO2014 / 068408, WO2014 / 070233, WO2014 / 071074, WO2014 / 073447, WO2014 / 078168, WO2014 / 080251, WO2014 / 085545, WO2014 / 085698, WO2 014 / 089335、WO2014 / 089379、WO2014 / 092804、WO2014 / 093631、WO2014 / 093675、WO2014 / 097318、WO2014 / 100438、WO2014 / 113641、WO2014 / 121033, WO2014 / 121211, WO2014 / 121235, WO2014 / 124227, WO2014 / 124322, WO2014 / 125465, WO2014 / 127200, WO2014 / 127785, WO2014 / 1343 11, WO2014 / 143240, WO2014 / 143241, WO2014 / 143242, WO2014 / 147611, WO2014 / 153117, WO2014 / 160183, WO2014 / 163684, WO2014 / 165506,WO2014 / 168953, WO2014 / 168986, WO2014 / 168993, WO2014 / 170704, WO2014 / 173886, WO2014 / 174060, WO2014 / 179528, WO2014 / 182521, WO2 014 / 193999、WO2014 / 194312、WO2014 / 194435、WO2014 / 197849、WO2014 / 197854、WO2014 / 197909、WO2014 / 201026、WO2015 / 010293、WO2015 / 012904, WO2015 / 013579, WO2015 / 014376, WO2015 / 014657, WO2015 / 015401, WO2015 / 015448, WO2015 / 023551, WO2015 / 031694, WO2015 / 0350 94, WO2015 / 037005, WO2015 / 038426, WO2015 / 038993, WO2015 / 040636, WO2015 / 047510, WO2015 / 048467, WO2015 / 049365, WO2015 / 054269, WO2015 / 065987, WO2015 / 069266, WO2015 / 069430, WO2015 / 073813, WO2015 / 082240, WO2015 / 082396, WO2015 / 084384, WO2015 / 084996, WO2 015 / 085208、WO2015 / 086593、WO2015 / 112311、WO2015 / 112847、WO2015 / 112854、WO2015 / 113176、WO2015 / 120150、WO2015 / 120254、WO2015 / 125640, WO2015 / 125641, WO2015 / 127685, WO2015 / 130416, WO2015 / 134607, WO2015 / 137777, WO2015 / 138377, WO2015 / 144184, WO2015 / 1484 15, WO2015 / 151078, WO2015 / 151080, WO2015 / 151081, WO2015 / 153401, WO2015 / 153820, WO2015 / 153877, WO2015 / 154064, WO2015 / 155753,WO2015 / 157376, WO2015 / 157471, WO2015 / 160845, WO2015 / 161192, WO2015 / 161247, WO2015 / 162563, WO2015 / 163817, WO2015 / 171589, WO2 015 / 175827、WO2015 / 179402、WO2015 / 181224、WO2015 / 181526、WO2015 / 188047、WO2015 / 195740、WO2016 / 003869、WO2016 / 004875、WO2016 / 004876, WO2016 / 007741, WO2016 / 012956, WO2016 / 022549, WO2016 / 022845, WO2016 / 022956, WO2016 / 025741, WO2016 / 026458, WO2016 / 0286 72, WO2016 / 032882, WO2016 / 033114, WO2016 / 036727, WO2016 / 040683, WO2016 / 040940, WO2016 / 044234, WO2016 / 045505, WO2016 / 046574, WO2016 / 057398, WO2016 / 059622, WO2016 / 061256, WO2016 / 064749, WO2016 / 064900, WO2016 / 067112, WO2016 / 069740, WO2016 / 073296, WO2 016 / 073733、WO2016 / 073845、WO2016 / 075211、WO2016 / 077264、WO2016 / 077505、WO2016 / 079067、WO2016 / 079331、WO2016 / 080626、WO2016 / 081364, WO2016 / 081603, WO2016 / 081884, WO2016 / 086847, WO2016 / 087340, WO2016 / 087952, WO2016 / 089208, WO2016 / 094837, WO2016 / 1038 67, WO2016 / 110228, WO2016 / 111751, WO2016 / 112870, WO2016 / 118697, WO2016 / 118951, WO2016 / 127022, WO2016 / 135513, WO2016 / 135633,WO2016 / 136641, WO2016 / 138542, WO2016 / 140714, WO2016 / 140884, WO2016 / 144637, WO2016 / 145014, WO2016 / 145096, WO2016 / 147031, WO2016 / 149668, WO2016 / 154019, WO2016 / 154629, WO2016 / 155655, WO2016 / 157219, WO2016 / 160833, WO2016 / 160881, WO2016 / 161391, WO2016 / 164641, WO2016 / 168784, WO2016 / 168885, WO2016 / 172427, WO2016 / 178224, WO2016 / 179108, WO2016 / 183398, WO2016 / 187393, WO2016 / 194688, WO2016 / 196062, WO2016 / 196894, WO2016 / 197032, WO2016 / 197114, WO2016 / 203478, WO2016 / 205459, WO2016 / 205551, WO2016 / 205566, WO2016 / 205738, WO2016 / 208591, WO2016 / 208592, WO2016 / 209795, WO2016 / 210108, WO2017 / 004144, WO2017 / 011312, WO2017 / 011371, WO2017 / 011411, WO2017 / 011590, WO2017 / 013593, WO2017 / 017063, WO2017 / 019804, WO2017 / 023667, WO2017 / 023994, WO2017 / 029407, WO2017 / 029601, WO2017 / 031261, WO2017 / 035116, WO2017 / 038607, WO2017 / 042634, WO2017 / 045603, WO2017 / 046602, WO2017 / 050979, WO2017 / 051249, WO2017 / 051254, WO2017 / 053243, WO2017 / 053464, WO2017 / 053920, WO2017 / 059357, WO2017 / 066726, WO2017 / 070092, WO2017 / 075052,WO2017 / 079746, WO2017 / 083151, WO2017 / 086392, WO2017 / 087685, WO2017 / 089890, WO2017 / 089894, WO2017 / 089895, WO2017 / 095823, WO2 017 / 095924、WO2017 / 100201、WO2017 / 101653、WO2017 / 106129、WO2017 / 106466、WO2017 / 106630、WO2017 / 118405、WO2017 / 118406、WO2017 / 118407, WO2017 / 120190, WO2017 / 132103, WO2017 / 132459, WO2017 / 136741, WO2017 / 137628, WO2017 / 139329, WO2017 / 139623, WO2017 / 1408 43, WO2017 / 141104, WO2017 / 149493, WO2017 / 150256, WO2017 / 151910, WO2017 / 157182, WO2017 / 161045, WO2017 / 162663, WO2017 / 165675, WO2017 / 165734, WO2017 / 172881, WO2017 / 176007, WO2017 / 177055, WO2017 / 177962, WO2017 / 180565, WO2017 / 181088, WO2017 / 181128, WO2 017 / 189279、WO2017 / 190059、WO2017 / 192573、WO2017 / 192863、WO2017 / 196806、WO2017 / 198124、WO2017 / 200787、WO2017 / 201278、WO2017 / 201528, WO2017 / 205741, WO2017 / 210058, WO2017 / 210566, WO2017 / 210963, WO2017 / 214491, WO2017 / 216791, WO2017 / 218544, WO2017 / 2212 70, WO2018 / 006756, WO2018 / 013993, WO2018 / 017426, WO2018 / 019284, WO2018 / 025168, WO2018 / 031408, WO2018 / 034356, WO2018 / 035413,WO2018 / 035423, WO2018 / 049155, WO2018 / 050027, WO2018 / 057649, WO2018 / 057712, WO2018 / 057973, WO2018 / 064683, WO2018 / 068021, WO2 018 / 068758、WO2018 / 083220、WO2018 / 084836、WO2018 / 086139、WO2018 / 089807、WO2018 / 090045、WO2018 / 094144、WO2018 / 094414、WO2018 / 098250, WO2018 / 102212, WO2018 / 102463, WO2018 / 102769, WO2018 / 103660, WO2018 / 103759, WO2018 / 106729, WO2018 / 106850, WO2018 / 1071 34, WO2018 / 111902, WO2018 / 112077, WO2018 / 112253, WO2018 / 112397, WO2018 / 114798, WO2018 / 125968, WO2018 / 136598, WO2018 / 136617, WO2018 / 140648, WO2018 / 140826, WO2018 / 141940, WO2018 / 142365, WO2018 / 144649, WO2018 / 148650, WO2018 / 153971, WO2018 / 156634, WO2 018 / 160758、WO2018 / 160909、WO2018 / 165482、WO2018 / 171164、WO2018 / 175994、WO2018 / 177287、WO2018 / 177369、WO2018 / 185526、WO2018 / 187236, WO2018 / 187287, WO2018 / 192505, WO2018 / 192550, WO2018 / 193413, WO2018 / 208700, WO2018 / 209239, WO2018 / 218087, WO2018 / 2268 46, WO2018 / 232334, WO2018 / 236190, WO2018 / 237354, WO2019 / 005634, WO2019 / 005635, WO2019 / 005636, WO2019 / 005756, WO2019 / 009813,WO2019 / 016208, WO2019 / 018238, WO2019 / 023435, WO2019 / 023621, WO2019 / 028055, WO2019 / 028133, WO2019 / 031614, WO2019 / 031615, WO2 019 / 032561、WO2019 / 034176、WO2019 / 034427、WO2019 / 038384、WO2019 / 038387、WO2019 / 038389、WO2019 / 038390、WO2019 / 040282、WO2019 / 040319, WO2019 / 051380, WO2019 / 051381, WO2019 / 055988, WO2019 / 065720, WO2019 / 070236, WO2019 / 075011, WO2019 / 075188, WO2019 / 0754 52, WO2019 / 075472, WO2019 / 078452, WO2019 / 082020, WO2019 / 090141, WO2019 / 090242, WO2019 / 096867, WO2019 / 099482, WO2019 / 102435, WO2019 / 105408, WO2019 / 108729, WO2019 / 114666, WO2019 / 125154, WO2019 / 127297, WO2019 / 127607, WO2019 / 133884, WO2019 / 136185, WO2 019 / 136309、WO2019 / 139979、WO2019 / 140188、WO2019 / 144012、WO2019 / 151827、WO2019 / 152667、WO2019 / 154120、WO2019 / 157003、WO2019 / 165200, WO2019 / 165285, WO2019 / 165340, WO2019 / 169138, WO2019 / 169143, WO2019 / 169160, WO2019 / 169168, WO2019 / 169181, WO2019 / 1751 13, WO2019 / 178055, WO2019 / 178443, WO2019 / 178487, WO2019 / 178490, WO2019 / 178494, WO2019 / 181939, WO2019 / 183117, WO2019 / 183145,WO2019 / 183209, WO2019 / 186497, WO2019 / 189419, WO2019 / 189732, WO2019 / 192979, WO2019 / 195665, WO2019 / 195770, WO2019 / 199621, WO2 019 / 199816、WO2019 / 200156、WO2019 / 200238、WO2019 / 215510、WO2019 / 222272、WO2019 / 222665、WO2019 / 223728、WO2019 / 227169、WO2019 / No. 232160, No. WO2019 / 236858, No. WO2019 / 237884, No. WO2019 / 238046, No. WO2019 / 241327, No. WO2019 / 242691, No. WO2020 / 006216, No. WO2020 / 006722, No. WO2020 / 0145 22, WO2020 / 023507, WO2020 / 023700, WO2020 / 023918, WO2020 / 030954, WO2020 / 032105, WO2020 / 033955, WO2020 / 035027, WO2020 / 036635, WO2020 / 038278, WO2020 / 042941, WO2020 / 043129, WO2020 / 043735, WO2020 / 044374, WO2020 / 045334, WO2020 / 055906, WO2020 / 056170, WO2 020 / 058458、WO2020 / 061007、WO2020 / 061106、WO2020 / 062951、WO2020 / 065408、WO2020 / 073345、WO2020 / 076849、WO2020 / 081148、WO2020 / 081460, WO2020 / 088667, WO2020 / 088702, WO2020 / 092344, WO2020 / 097306, WO2020 / 097336, WO2020 / 098658, WO2020 / 102404, WO2020 / 1025 29, WO2020 / 102852, WO2020 / 108600, WO2020 / 112627, WO2020 / 112935, WO2020 / 118265, WO2020 / 118605, WO2020 / 120785, WO2020 / 123275,WO2020 / 125546, WO2020 / 126064, WO2020 / 146906, WO2020 / 148641, WO2020 / 154672, WO2020 / 155017, WO2020 / 156189, WO2020 / 156513, WO2 020 / 160009、WO2020 / 160222、WO2020 / 166592、WO2020 / 167868、WO2020 / 168106、WO2020 / 169004、WO2020 / 172492、WO2020 / 176497、WO2020 / 180121, WO2020 / 183307, WO2020 / 190984, WO2020 / 198293, WO2020 / 207974, WO2020 / 210366, WO2020 / 210689, WO2020 / 210692, WO2020 / 2106 94, WO2020 / 211860, WO2020 / 220085, WO2020 / 222573, WO2020 / 228265, WO2020 / 228656, WO2020 / 232119, WO2020 / 232701, WO2020 / 233174, WO2020 / 233602, WO2020 / 233723, WO2020 / 236901, WO2020 / 241340, WO2020 / 243745, WO2020 / 244657, WO2020 / 247576, WO2020 / 252401, WO2 020 / 257998、WO2020 / 258893、WO2020 / 259258、WO2020 / 259601、WO2021 / 004482、WO2021 / 009759、WO2021 / 016182、WO2021 / 018240、WO2021 / 021523, WO2021 / 022121, WO2021 / 023973, WO2021 / 028362, WO2021 / 034194, WO2021 / 041672, WO2021 / 044208, WO2021 / 047646, WO2021 / 0484 69, WO2021 / 052402, WO2021 / 055705, WO2021 / 060453, WO2021 / 067761, WO2021 / 067776, WO2021 / 067820, WO2021 / 067861, WO2021 / 072350,WO2021 / 081244, WO2021 / 092672, WO2021 / 097046, WO2021 / 108853, WO2021 / 113625, WO2021 / 113643, WO2021 / 113644, WO2021 / 113651, WO2 021 / 113657、WO2021 / 113740、WO2021 / 123013、WO2021 / 150792、WO2021 / 151984、WO2021 / 155580、WO2021 / 158780、WO2021 / 163484、WO2021 / 164765, WO2021 / 173773, WO2021 / 174034, WO2021 / 174113, WO2021 / 178253, WO2021 / 178396, WO2021 / 194298, WO2021 / 202642, WO2021 / 2027 26, WO2021 / 202770, WO2021 / 202900, WO2021 / 209007, WO2021 / 212638, WO2021 / 213504, WO2021 / 216898, WO2021 / 221330, WO2021 / 222138, WO2021 / 226033, WO2021 / 226092, WO2021 / 226368, WO2021 / 226584, WO2021 / 226849, WO2021 / 231568, WO2021 / 231988, WO2021 / 234550, WO2 021 / 248065、WO2021 / 249228、WO2021 / 252889、WO2021 / 259506、WO2021 / 259507、WO2021 / 259944、WO2021 / 260438、WO2021 / 261992、WO2021 / 261995, WO2021 / 261998, WO2021 / 262843, WO2021 / 262998, WO2022 / 000868, WO2022 / 006083, WO2022 / 006685, WO2022 / 015656, WO2022 / 0269 15, WO2022 / 029220, WO2022 / 029334, WO2022 / 031605, WO2022 / 033578, WO2022 / 035818, WO2022 / 035888, WO2022 / 037665, WO2022 / 040603,WO2022 / 040635, WO2022 / 042583, WO2022 / 050300, WO2022 / 052823, WO2022 / 061251, WO2022 / 064191, WO2022 / 075713, WO2022 / 078259, WO2 022 / 078260、WO2022 / 078524、WO2022 / 081895、WO2022 / 082066、WO2022 / 082068、WO2022 / 083694、WO2022 / 084560、WO2022 / 087243、WO2022 / 100535, WO2022 / 103840, WO2022 / 103856, WO2022 / 109334, WO2022 / 111616, WO2022 / 112356, WO2022 / 112865, WO2022 / 112866, WO2022 / 1147 36, WO2022 / 115488, WO2022 / 116943, WO2022 / 120084, WO2022 / 120490, WO2022 / 121981, WO2022 / 124898, WO2022 / 132652, WO2022 / 135560, WO2022 / 138944, WO2022 / 153211, WO2022 / 159582, WO2022 / 164316, WO2022 / 165403, WO2022 / 166762, WO2022 / 167682, WO2022 / 169329, WO2 022 / 170971、WO2022 / 171115、WO2022 / 174834、WO2022 / 175402、WO2022 / 175595、WO2022 / 180581、WO2022 / 182993、WO2022 / 185315、WO2022 / 187584, WO2022 / 194257, WO2022 / 195066, WO2022 / 197877, WO2022 / 198231, WO2022 / 198232, WO2022 / 198325, WO2022 / 200347, WO2022 / 2042 69, WO2022 / 207924, WO2022 / 211508, WO2022 / 214598, WO2022 / 216616, WO2022 / 217022, WO2022 / 225737, WO2022 / 226317, WO2022 / 229932,WO2022 / 235585, WO2022 / 240966, WO2022 / 241418, WO2022 / 245061, WO2022 / 246576, WO2022 / 247190, WO2022 / 253284, WO2022 / 256371, WO2 022 / 261310、WO2022 / 262516、WO2022 / 265880、WO2022 / 271671、WO2023 / 000131、WO2023 / 002431、WO2023 / 004349、WO2023 / 005953、WO2023 / 011631, WO2023 / 020605, WO2023 / 025138, WO2023 / 028605, WO2023 / 030064, WO2023 / 030364, WO2023 / 033990, WO2023 / 037254, WO2023 / 0517 96, WO2023 / 052783, WO2023 / 052971, WO2023 / 056378, WO2023 / 057813, WO2023 / 059600, WO2023 / 060283, WO2023 / 061224, WO2023 / 065137, WO2023 / 066865, WO2023 / 069727, WO2023 / 072143, WO2023 / 072755, WO2023 / 072934, WO2023 / 076998, WO2023 / 079294, WO2023 / 081727, WO2 023 / 083381、WO2023 / 083919、WO2023 / 086951、WO2023 / 088086、WO2023 / 091726、WO2023 / 092089、WO2023 / 092099、WO2023 / 097604、WO2023 / 098889, WO2023 / 100829, WO2023 / 101928, WO2023 / 102875, WO2023 / 104188, WO2023 / 109836, WO2023 / 109965, WO2023 / 121447, WO2023 / 1287 08, WO2023 / 130012, WO2023 / 134706, WO2023 / 137026, WO2023 / 141560, WO2023 / 141855, WO2023 / 147063, WO2023 / 151679, WO2023 / 152336,WO2023 / 153442, WO2023 / 154907, WO2023 / 161291, WO2023 / 163227, WO2023 / 163234, WO2023 / 164610, WO2023 / 167238, WO2023 / 172880, WO2 023 / 173026、WO2023 / 173121、WO2023 / 175366、WO2023 / 177233、WO2023 / 177629、WO2023 / 178179、WO2023 / 178202、WO2023 / 178289、WO2023 / 178451, WO2023 / 178452, WO2023 / 178964, WO2023 / 180484, WO2023 / 180489, WO2023 / 180490, WO2023 / 183932, WO2023 / 190827, WO2023 / 1929 61, WO2023 / 192969, WO2023 / 194441, WO2023 / 194800, WO2023 / 196742, WO2023 / 196761, WO2023 / 198884, WO2023 / 201267, WO2023 / 201399, WO2023 / 201432, WO2023 / 204631, WO2023 / 205344, WO2023 / 205807, WO2023 / 207475, WO2023 / 207773, WO2023 / 207875, WO2023 / 212704, WO2 023 / 212725、WO2023 / 214849、WO2023 / 216956、WO2023 / 217064、WO2023 / 217227、WO2023 / 221320、WO2023 / 223097、WO2023 / 227006、WO2023 / 227660, WO2023 / 230624, WO2023 / 232048, WO2023 / 235716, WO2023 / 235772, WO2023 / 237050, WO2023 / 239710, WO2023 / 239803, WO2023 / 2402 91, WO2023 / 249473, WO2023 / 249974, WO2023 / 275112, WO2023 / 280227, WO2024 / 001669, WO2024 / 006542, WO2024 / 013354, WO2024 / 014837,WO2024 / 015229, WO2024 / 015830, WO2024 / 020355, WO2024 / 020491, WO2024 / 020734, WO2024 / 022167, WO2024 / 025845, WO2024 / 033667, WO2024 / 044570, WO2024 / 047639, WO2024 / 049220, WO2024 / 050031, WO2024 / 051502, WO2024 / 052333, WO2024 / 054089, WO2024 / 054469, WO2024 / 054821, WO2024 / 059236, WO2024 / 062319, WO2024 / 064024, WO2024 / 064968, WO2024 / 068976, WO2024 / 081345, WO2024 / 082051, WO2024 / 082055, WO2024 / 082056, WO2024 / 083161, WO2024 / 083162, WO2024 / 083925, WO2024 / 083926, WO2024 / 083953, WO2024 / 088388, WO2024 / 092016, WO2024 / 095012, WO2024 / 097559, WO2024 / 100200, WO2024 / 100285, WO2024 / 102956, WO2024 / 105197, WO2024 / 105610, WO2024 / 107014, WO2024 / 123896, WO2024 / 125627, WO2024 / 126735, WO2024 / 127398, WO2024 / 130161, WO2024 / 133845, WO2024 / 138000, WO2024 / 140443, WO2024 / 140447, WO2024 / 140449, WO2024 / 140450, WO2024 / 140933, WO2024 / 148241, WO2024 / 148431, WO2024 / 149345, WO2024 / 151890, WO2024 / 155728, WO2024 / 159067, WO2024 / 159079, WO2024 / 159081, WO2024 / 159088, WO2024 / 159103, WO2024 / 163630, WO2024 / 165045, WO2024 / 173761,WO2024 / 175069, WO2024 / 182311, WO2024 / 187006, WO2024 / 189269, WO2024 / 189620, WO2024 / 191826, WO2024 / 193635, WO2024 / 194851, WO2 024 / 199337、WO2024 / 205474、WO2024 / 206480、WO2024 / 206669、WO2024 / 208314、WO2024 / 208354、WO2024 / 208359、WO2024 / 210687、WO2024 / 212922, WO2024 / 213119, WO2024 / 215624, WO2024 / 218697, WO2024 / 225848, WO2024 / 226410, WO2024 / 227119, WO2024 / 227432, WO2024 / 2291 00, WO2024 / 230646, WO2024 / 230678, WO2024 / 230752, WO2024 / 231929, WO2024 / 231930, WO2024 / 235105, WO2024 / 238454, WO2024 / 239281, WO2024 / 240227, WO2024 / 243545, WO2024 / 244006, WO2024 / 251149, WO2024 / 251743, WO2024 / 255740, WO2024 / 259300, WO2024 / 261742, WO2 025 / 012080、WO2025 / 014541、WO2025 / 014830、WO2025 / 016467、WO2025 / 019776、WO2025 / 019780、WO2025 / 019787、WO2025 / 021035、WO2025 / 021152, WO2025 / 027194, WO2025 / 029578, WO2025 / 036876, WO2025 / 038395, WO2025 / 038638, WO2005 / 081711, WO2024 / 116094, WO2024 / 0237 50, WO2023 / 228095, WO2023 / 218378, WO2023 / 209591, WO2023 / 176881, WO2023 / 100829, WO2022 / 102695, WO2022 / 102634, WO2022 / 014698,WO2020 / 240467, WO2020 / 130125, WO2020 / 122034, WO2020 / 111234, WO2020 / 059772, WO2020 / 040245, WO2020 / 031936, WO2020 / 027100, WO2 020 / 022475、WO2020 / 022363、WO2019 / 230645、WO2019 / 219891、WO2019 / 044947、WO2019 / 039483、WO2018 / 169092、WO2018 / 159582、WO2018 / 110515, WO2018 / 066626, WO2018 / 060714, WO2017 / 137503, WO2017 / 056012, WO2016 / 133194, WO2016 / 072440, WO2016 / 023894, WO2015 / 1559 98, WO2015 / 155976, WO2015 / 115091, WO2015 / 098099, WO2014 / 061277, WO2014 / 057687, WO2013 / 163229, WO2011 / 104981, WO2011 / 038159, WO2009 / 084614, WO2025 / 005240, WO2024 / 248123, WO2024 / 219442, WO2024 / 214685, WO2024 / 210162, WO2024 / 204679, WO2024 / 204629, WO2 024 / 190815、WO2024 / 158047、WO2024 / 048490、WO2023 / 219072、WO2023 / 176863、WO2023 / 176862、WO2023 / 167238、WO2023 / 136342、WO2023 / 100829, WO2023 / 074843, WO2023 / 068226, WO2022 / 244825, WO2022 / 244815, WO2022 / 244801, WO2022 / 210485, WO2022 / 196719, WO2022 / 1913 13, WO2022 / 191312, WO2022 / 163846, WO2022 / 102695, WO2022 / 102694, WO2022 / 102652, WO2022 / 065344, WO2021 / 251453, WO2021 / 241730,WO2021 / 200857, WO2021 / 177438, WO2021 / 112251, WO2021 / 095838, WO2021 / 079958, WO2021 / 025148, WO2021 / 020282, WO2021 / 006268, WO2 020 / 240467、WO2020 / 196712、WO2020 / 196475、WO2020 / 196474、WO2020 / 179859、WO2020 / 153390、WO2020 / 130125、WO2020 / 122034、WO2020 / 116662, WO2020 / 111234, WO2020 / 100954, WO2020 / 095922, WO2020 / 095921, WO2020 / 050406, WO2020 / 032153, WO2020 / 013170, WO2020 / 0102 52, WO2020 / 009156, WO2019 / 245012, WO2019 / 244107, WO2019 / 240164, WO2019 / 235553, WO2019 / 230869, WO2019 / 230645, WO2019 / 219891, WO2019 / 172165, WO2019 / 156115, WO2019 / 070013, WO2019 / 065928, WO2019 / 054427, WO2019 / 049933, WO2019 / 044947, WO2019 / 039483, WO2 019 / 023198、WO2019 / 022187、WO2019 / 017338、WO2019 / 009407、WO2019 / 009265、WO2019 / 004487、WO2018 / 235966、WO2018 / 212136、WO2018 / 181656, WO2018 / 159582, WO2018 / 147245, WO2018 / 139464, WO2018 / 135556, WO2018 / 135501, WO2018 / 123925, WO2018 / 117244, WO2018 / 1172 37, WO2018 / 110515, WO2018 / 097166, WO2018 / 097162, WO2018 / 070390, WO2018 / 066492, WO2018 / 047894, WO2018 / 003983, WO2017 / 172764,WO2017 / 137503, WO2017 / 069289, WO2017 / 051888, WO2017 / 018499, WO2017 / 010559, WO2017 / 002776, WO2016 / 184909, WO2016 / 171107, WO2 016 / 167276、WO2016 / 167236、WO2016 / 148115、WO2016 / 121908、WO2016 / 056673、WO2016 / 052697、WO2016 / 023894、WO2016 / 021674、WO2016 / 017693, WO2016 / 006706, WO2015 / 155998, WO2015 / 155976, WO2015 / 146132, WO2015 / 141616, WO2015 / 129812, WO2015 / 125956, WO2015 / 1150 91, WO2015 / 108175, WO2015 / 098099, WO2015 / 053407, WO2015 / 048802, WO2015 / 048793, WO2015 / 046505, WO2015 / 041318, WO2015 / 019976, WO2015 / 005473, WO2015 / 005253, WO2014 / 140210, WO2014 / 133071, WO2014 / 115797, WO2014 / 069672, WO2014 / 038606, WO2014 / 024914, WO2 013 / 172379、WO2013 / 163229、WO2013 / 161895、WO2013 / 160879、WO2013 / 154206、WO2013 / 147213、WO2013 / 147212、WO2013 / 115280、WO2013 / 080934, WO2013 / 027680, WO2013 / 002248, WO2012 / 160464, WO2012 / 147713, WO2012 / 121361, WO2012 / 074038, WO2012 / 057288, WO2012 / 0454 81, WO2012 / 026434, WO2012 / 005378, WO2012 / 002274, WO2011 / 115264, WO2011 / 111824, WO2011 / 102404, WO2011 / 087102, WO2011 / 052715,WO2011 / 013728, WO2011 / 013656, WO2010 / 117011, WO2010 / 093055, WO2010 / 082612, WO2010 / 044401, WO2010 / 016490, WO2010 / 001909, WO2009 / 151069, WO2009 / 119537, WO2009 / 075376, WO2009 / 057813, WO2009 / 048072, WO2009 / 0349 76, WO2009 / 028639, WO2009 / 020093, WO2008 / 072655, WO2008 / 053962, WO2008 / 016133, WO2007 / 132949, WO2007 / 129745, WO2024 / 243450, WO2024 / 042497, WO2022 / 056281, WO2021 / 127499, WO2020 / 223538, WO2019 / 246513, WO2019 / 191630, WO2019 / 090090, WO2019 / 090088, WO2019 / 090085, WO2019 / 090082, WO2019 / 090081, WO2019 / 090078, WO2019 / 090076, WO2019 / 090074, WO2019 / 090069, WO2018 / 188047, WO2018 / 107116, WO2018 / 107109, WO2018 / 068283, WO2 018 / 068282、WO2018 / 053106、WO2017 / 214462、WO2017 / 214458、WO2017 / 214456、WO2017 / 214339、WO2017 / 214322、WO2017 / 214301、WO2017 / 214282、WO2017 / 214233、WO2017 / 205745、WO2017 / 205742、WO2017 / 205738、WO2017 / 201442、 WO2017 / 193096, WO2017 / 193063, WO2017 / 193041, WO2017 / 193034, WO2017 / 193030, WO2017 / 184942, WO2017 / 177955, WO2017 / 112829, WO2017 / 112811, WO2017 / 112803, WO2017 / 096163,WO2017 / 095808, WO2017 / 095805, WO2017 / 087800, WO2017 / 031458, WO2017 / 011342, WO2016 / 196314, WO2016 / 044770, WO2016 / 004389, WO2 015 / 200635、WO2015 / 191934、WO2015 / 143382、WO2015 / 119712、WO2015 / 089075、WO2015 / 085925、WO2015 / 073884、WO2015 / 058173、WO2015 / 051379, WO2015 / 035044, WO2014 / 210425, WO2014 / 210209, WO2014 / 206150, WO2014 / 176415, WO2014 / 165143, WO2014 / 165127, WO2014 / 1647 80, WO2014 / 164771, WO2014 / 160028, WO2014 / 160017, WO2014 / 159999, WO2014 / 158528, WO2014 / 152199, WO2014 / 139328, WO2014 / 145000, WO2014 / 144960, WO2014 / 144935, WO2014 / 144299, WO2014 / 144280, WO2014 / 143765, WO2014 / 116846, WO2014 / 106015, WO2014 / 106001, WO2 014 / 105810、WO2014 / 089209、WO2014 / 085654、WO2014 / 071074、WO2014 / 047222、WO2014 / 028381、WO2013 / 188381、WO2013 / 170118、WO2013 / 170115, WO2013 / 170113, WO2013 / 170112, WO2013 / 158952, WO2013 / 126656, WO2013 / 102042, WO2013 / 101993, WO2013 / 101972, WO2013 / 0976 01, WO2013 / 090666, WO2013 / 063095, WO2013 / 059485, WO2013 / 055897, WO2013 / 055895, WO2013 / 013031, WO2012 / 121758, WO2024 / 263630,WO2024 / 184349, WO2023 / 079057, WO2023 / 023489, WO2023 / 015278, WO2017 / 015124, WO2017 / 015008, WO2003 / 076567, WO2002 / 090993, WO2 002 / 020751、WO2023 / 201185、WO2022 / 232333、WO2022 / 011075、WO2020 / 210384、WO2020 / 181283、WO2020 / 135483、WO2019 / 101183、WO2019 / 101182, WO2019 / 001419, WO2018 / 009525, WO2017 / 053864, WO2016 / 127025, WO2016 / 081679, WO2016 / 057500, WO2015 / 061751, WO2015 / 0612 47, WO2014 / 093230, WO2014 / 085528, WO2014 / 031936, WO2014 / 008312, WO2013 / 096194, WO2013 / 078145, WO2013 / 042006, WO2013 / 040044, WO2013 / 039988, WO2012 / 177892, WO2012 / 170976, WO2012 / 143726, WO2012 / 135113, WO2012 / 061409, WO2012 / 005805, WO2011 / 140325, WO2 011 / 140324、WO2011 / 138751、WO2011 / 135376、WO2011 / 098451、WO2011 / 086053、WO2011 / 062965、WO2011 / 060281、WO2010 / 151495、WO2010 / 132725, WO2010 / 077678, WO2010 / 054266, WO2010 / 048278, WO2010 / 045580, WO2010 / 038165, WO2010 / 021770, WO2009 / 151495, WO2009 / 1404 29, WO2009 / 140423, WO2009 / 140421, WO2009 / 137391, WO2009 / 098448, WO2009 / 091550, WO2009 / 055418, WO2009 / 047563, WO2009 / 036082,WO2009 / 016460, WO2008 / 157358, WO2008 / 154333, WO2008 / 115478, WO2008 / 094896, WO2008 / 073922, WO2008 / 073921, WO2008 / 060866, WO2 008 / 036776、WO2008 / 032162、WO2007 / 132308、WO2007 / 109211、WO2007 / 075567、WO2007 / 048088、WO2007 / 024754、WO2007 / 023382、WO2007 / 000676, WO2006 / 138155, WO2006 / 135721, WO2006 / 135629, WO2006 / 117669, WO2006 / 101937, WO2006 / 081327, WO2006 / 076442, WO2006 / 0556 76, WO2006 / 040646, WO2006 / 031330, WO2005 / 121124, WO2005 / 115145, WO2005 / 113494, WO2005 / 072731, WO2005 / 071096, WO2005 / 009435, WO2004 / 094463, WO2004 / 063334, WO2004 / 048598, WO2003 / 008410, WO2002 / 095000, WO2002 / 060919, WO2002 / 057414, WO2002 / 003997, WO2 001 / 064251、WO1999 / 059565、WO2025 / 042953、WO2025 / 031307、WO2025 / 015281、WO2024 / 245382、WO2024 / 231224、WO2024 / 230742、WO2024 / 199462, WO2024 / 199432, WO2024 / 197082, WO2024 / 182454, WO2024 / 178155, WO2024 / 165057, WO2024 / 163920, WO2024 / 163630, WO2024 / 1601 76, WO2024 / 147097, WO2024 / 133763, WO2024 / 102523, WO2024 / 067811, WO2024 / 052333, WO2024 / 047654, WO2024 / 022165, WO2024 / 002330,WO2023 / 280092, WO2023 / 247731, WO2023 / 247729, WO2023 / 232144, WO2023 / 225359, WO2023 / 201234, WO2023 / 180485, WO2023 / 180484, WO2 023 / 174213、WO2023 / 173074、WO2023 / 143347、WO2023 / 138635、WO2023 / 121447、WO2023 / 118961、WO2023 / 109953、WO2023 / 109836、WO2023 / 107538, WO2023 / 077126, WO2023 / 017191, WO2022 / 205111, WO2022 / 187272, WO2022 / 187270, WO2022 / 161385, WO2022 / 105873, WO2022 / 0937 93, WO2022 / 078566, WO2022 / 068898, WO2019 / 218944, WO2019 / 055525, WO2018 / 095422, WO2017 / 222310, WO2017 / 214151, WO2011 / 073091, WO2008 / 130439, WO2008 / 079972, WO2008 / 079968, WO2008 / 070150, WO2008 / 048375, WO2008 / 045604, WO2007 / 064872, WO2007 / 059202, WO2 006 / 064517、WO2005 / 110994、WO2004 / 078747、WO2009 / 079790、WO2009 / 076170、WO2009 / 074677、WO2009 / 070609、WO2009 / 061770、WO2009 / 059201, WO2009 / 055343, WO2009 / 054332, WO2009 / 049305, WO2009 / 038779, WO2009 / 037001, WO2009 / 030239, WO2009 / 026446, WO2009 / 0262 74, WO2009 / 020601, WO2009 / 018092, WO2009 / 012280, WO2009 / 012096, WO2009 / 008992, WO2009 / 005673, WO2009 / 002808, WO2009 / 002807,WO2009 / 002806, WO2009 / 002719, WO2008 / 154927, WO2008 / 153207, WO2008 / 141111, WO2008 / 141110, WO2008 / 141107, WO2008 / 134761, WO2 008 / 134679、WO2008 / 134088、WO2008 / 133866、WO2008 / 127656、WO2008 / 124735、WO2008 / 121949、WO2008 / 120098、WO2008 / 115556、WO2008 / 115499, WO2008 / 109432, WO2008 / 109163, WO2008 / 109154, WO2008 / 106491, WO2008 / 103470, WO2008 / 100922, WO2008 / 098789, WO2008 / 0987 88, WO2008 / 098138, WO2008 / 094834, WO2008 / 094319, WO2008 / 087643, WO2008 / 087488, WO2008 / 083027, WO2008 / 082887, WO2008 / 082602, WO2008 / 081914, WO2008 / 079735, WO2008 / 079346, WO2008 / 079240, WO2008 / 076278, WO2008 / 076257, WO2008 / 073817, WO2008 / 067027, WO2 008 / 064133、WO2008 / 063663、WO2008 / 062466、WO2008 / 051421、WO2008 / 049633、WO2008 / 049105、WO2008 / 045184、WO2008 / 043576、WO2008 / 039254, WO2008 / 034776, WO2008 / 020827, WO2008 / 020045, WO2008 / 020024, WO2008 / 014574, WO2008 / 013987, WO2008 / 013840, WO2008 / 0125 11, WO2008 / 007122, WO2008 / 006883, WO2008 / 006535, WO2008 / 003317, WO2008 / 001115, WO2008 / 001101, WO2007 / 144384, WO2007 / 139569,WO2007 / 134037, WO2007 / 129062, WO2007 / 126122, WO2007 / 124383, WO2007 / 123995, WO2007 / 123892, WO2007 / 114549, WO2007 / 113687, WO2 007 / 112193、WO2007 / 109184、WO2007 / 107305、WO2007 / 106356、WO2007 / 106192、WO2007 / 105027、WO2007 / 098091、WO2007 / 098090、WO2007 / 098089, WO2007 / 097934, WO2007 / 096395, WO2007 / 096393, WO2007 / 095389, WO2007 / 092496, WO2007 / 089864, WO2007 / 080114, WO2007 / 0760 87, WO2007 / 075572, WO2007 / 067417, WO2007 / 064945, WO2007 / 064759, WO2007 / 062138, WO2007 / 056215, WO2007 / 056214, WO2007 / 056208, WO2007 / 054551, WO2007 / 046893, WO2007 / 041546, WO2007 / 041453, WO2007 / 041397, WO2007 / 039404, WO2007 / 039403, WO2007 / 038658, WO2 007 / 027344、WO2007 / 024940、WO2007 / 019678、WO2006 / 136837、WO2006 / 135479、WO2006 / 129163、WO2006 / 125815、WO2006 / 125813、WO2006 / 124021, WO2006 / 121522, WO2006 / 121521, WO2006 / 121518, WO2006 / 113679, WO2006 / 112930, WO2006 / 109091, WO2006 / 107903, WO2006 / 1077 86, WO2006 / 107617, WO2006 / 105979, WO2006 / 105237, WO2006 / 099015, WO2006 / 097474, WO2006 / 088906, WO2006 / 086484, WO2006 / 084904,WO2006 / 084869, WO2006 / 083260, WO2006 / 081337, WO2006 / 081331, WO2006 / 079645, WO2006 / 079644, WO2006 / 079478, WO2006 / 078711, WO2 006 / 077425、WO2006 / 077424、WO2006 / 075012、WO2006 / 073430、WO2006 / 070024、WO2006 / 070023、WO2006 / 069063、WO2006 / 065780、WO2006 / 065234, WO2006 / 057429, WO2006 / 055676, WO2006 / 052767, WO2006 / 048892, WO2006 / 042146, WO2006 / 037230, WO2006 / 034128, WO2006 / 0273 46, WO2006 / 018182, WO2006 / 015318, WO2006 / 014361, WO2006 / 009805, WO2005 / 123676, WO2005 / 117980, WO2005 / 115391, WO2005 / 112919, WO2005 / 107712, WO2005 / 094830, WO2005 / 094818, WO2005 / 094802, WO2005 / 092097, WO2005 / 087221, WO2005 / 082889, WO2005 / 082422, WO2 005 / 082353、WO2005 / 082023、WO2005 / 060663、WO2005 / 052143、WO2005 / 051444、WO2005 / 051452、WO2005 / 051432、WO2005 / 051430、WO2005 / 051229, WO2005 / 049105, WO2005 / 049091, WO2005 / 048822, WO2005 / 046747, WO2005 / 046746, WO2005 / 046637, WO2005 / 046599, WO2005 / 0450 55, WO2005 / 044365, WO2005 / 044302, WO2005 / 019435, WO2005 / 032492, WO2005 / 031002, WO2005 / 028539, WO2005 / 027907, WO2005 / 026328,WO2005 / 025508, WO2005 / 023294, WO2005 / 020958, WO2005 / 019247, WO2005 / 018683, WO2005 / 007136, WO2005 / 003172, WO2005 / 003162, WO2 005 / 000220、WO2004 / 111603、WO2004 / 110455、WO2004 / 103358、WO2004 / 098569、WO2004 / 096224、WO2004 / 093807、WO2004 / 089423、WO2004 / 089294, WO2004 / 087075, WO2004 / 085418, WO2004 / 085386, WO2004 / 080478, WO2004 / 076675, WO2004 / 073656, WO2004 / 072641, WO2004 / 0692 74, WO2004 / 069201, WO2004 / 060405, WO2004 / 060346, WO2004 / 056862, WO2004 / 054624, WO2004 / 050867, WO2004 / 050041, WO2004 / 043493, WO2004 / 038020, WO2004 / 037751, WO2004 / 032828, WO2004 / 020457, WO2004 / 018000, WO2004 / 014882, WO2004 / 014314, WO2004 / 010900, WO2 004 / 009023、WO2004 / 008101、WO2004 / 006847、WO2004 / 004727、WO2003 / 103822、WO2003 / 103596、WO2003 / 098220、WO2003 / 093297、WO2003 / 090785, WO2003 / 090680, WO2003 / 089006, WO2003 / 086470, WO2003 / 082301, WO2003 / 080039, WO2003 / 074586, WO2003 / 074551, WO2003 / 0701 66, WO2003 / 066830, WO2003 / 059988, WO2003 / 057158, WO2003 / 055890, WO2003 / 055866, WO2003 / 055489, WO2003 / 053446, WO2003 / 050295,WO2003 / 044482, WO2003 / 043631, WO2003 / 043583, WO2003 / 041643, WO2003 / 037860, WO2003 / 031579, WO2003 / 026577, WO2003 / 024967, WO2 003 / 020262、WO2003 / 016500、WO2003 / 015698、WO2003 / 009881、WO2003 / 009777、WO2003 / 000928、WO2002 / 100353、WO2002 / 096367、WO2002 / 092826, WO2002 / 089772, WO2002 / 088714, WO2002 / 088327, WO2002 / 085459, WO2002 / 083180, WO2002 / 083067, WO2002 / 080849, WO2002 / 0770 36, WO2002 / 076486, WO2002 / 076448, WO2002 / 072151, WO2002 / 065136, WO2002 / 058694, WO2002 / 056790, WO2002 / 053138, WO2002 / 051862, WO2002 / 051444, WO2002 / 047611, WO2002 / 045737, WO2002 / 022159, WO2002 / 020500, WO2002 / 013843, WO2002 / 009720, WO2002 / 004409, WO2 002 / 0001168、WO2001 / 097855、WO2001 / 095945、WO2001 / 093836、WO2001 / 092584、WO2001 / 091807、WO2001 / 087307、WO2001 / 080843、WO2001 / 079187, WO2001 / 074402, WO2001 / 074401, WO2001 / 072721, WO2001 / 066708, WO2001 / 064687, WO2001 / 064252, WO2001 / 049284, WO2001 / 0492 68, WO2001 / 039815, WO2001 / 036007, WO2001 / 028542, WO2001 / 025407, WO2001 / 024763, WO2001 / 018013, WO2001 / 017563, WO2001 / 005437,WO2000 / 073340, WO2000 / 069390, WO2000 / 062075, WO2000 / 061597, WO2000 / 050016, WO2000 / 005415, WO1999 / No. 065515, WO1999 / 049901, WO1999 / 046371, WO1999 / 030684, WO1999 / 029329, WO1999 / 003976, WO1998 / 054966 , WO1998 / 019705, WO1997 / 044492, WO1997 / 044336, WO1997 / 044026, WO1997 / 041898, WO1997 / 009998, WO1996 / 010585, WO1996 / 007322, WO1996 / 003984, WO1996 / 001127, WO1992 / 020813, and WO1991 / 016904.

[0105] Specific examples of the composite compound of a topoisomerase I inhibitor and a linker that can be preferably used in the present invention include CL2A-SN38, CL2-SN38, and PAB-Lys(MMT)-oxydiacetamide-PEG. ８ -N3-SN38, MC-SN38, PH-HG-005-5, Mc-VC-PAB-SN38, CL2E-SN38, CL2E-SN38 TFA, CL2-MMT-SN38, deruxtecan (DXd: a compound in which the linker Gly-Gly-Phe-Gly is attached to MAAAA-1181a), which is an exatecan derivative, and propargyl-PEG ４ -GGFG-DXd, DBCO-PEG ４ -GGFG-Dxd, Br-Val-Ala-NH ２-bicyclo[1.1.1]pentane-7-MAD-MDCPT, MC-Gly-Gly-Phe-Gly-(R)-Cyclopropane-Exatecan, MC-Gly-Gly-Phe-Gly-(S)-Cyc lopropane-Exatecan, Val-Cit-PAB-Exatecan, MC-Val-Cit-PAB-Exatecan, Val-Ala-PABC-Exatecan, Val-Ala-PABC-Exatecan trifluoroacetate, MC-VA-PAB-Exatecan, MC-GGFG-Exatecan, MC-Gly-Gly-Phe-Gly-Cyclobutanecarboxylic-Exatecan, PNU-EDA-Gly5, or derivatives thereof.

[0106] <Conjugate of anti-CAPRIN-1 antibody and topoisomerase I inhibitor> In the present invention, the bond between the anti-CAPRIN-1 antibody and the topoisomerase I inhibitor in the conjugate thereof is not particularly limited as long as it is a bond that can maintain anti-cancer activity, but a bond that forms a linker structure between the anti-CAPRIN-1 antibody and the topoisomerase I inhibitor is preferred.

[0107] Here, the linker refers to a compound capable of binding an anti-CAPRIN-1 antibody to a topoisomerase I inhibitor. Various known linkers may be used, or the structure of the topoisomerase I inhibitor may be chemically modified to correspond to a linker and then bound to the antibody.

[0108] The type of linker and the details of the binding method can be determined in accordance with known methods (see, for example, Greg T. Hermanson, Bioconjugate Techniques, Third Edition, WO2004 / 010957 and WO2014 / 012479).

[0109] In embodiments of the invention, reactive groups attached to the anti-CAPRIN-1 antibody, topoisomerase I inhibitor, and linker include:

[0110] Unless specifically chemically modified, reactive groups present in the amino acid sequence of an antibody or in glycoproteins modified with amino acids include primary amines (ε-amino), carboxyls, thiols (sulfhydryls), carbonyls (ketones or aldehydes), and hydroxyls. Primary amines are found at the N-terminus of polypeptides and in the side chains of lysine residues. They are positively charged under physiological conditions and are usually located on the outside of proteins, allowing them to be used for conjugation without altering the protein structure. Carboxyls are found at the C-terminus of polypeptides and in the side chains of aspartic acid and glutamic acid. Sulfhydryls are found in the side chains of cysteine ​​and form disulfide bonds that maintain the higher-order structure of proteins. Ketones or aldehydes are generated in glycoproteins by oxidizing glycosyl groups with sodium metaperiodate.

[0111] The conjugates of the present invention are prepared by attaching a drug to a linker attached to the reactive group on the antibody, by attaching the antibody to a linker attached to the drug, or by attaching the drug directly to the antibody.

[0112] Reactive groups attached to the linker and topoisomerase inhibitor include:

[0113] Reactive groups that can react with amines include N-hydroxysuccinimide (NHS) esters, imidoesters, pentafluorophenyl esters, hydroxymethylphosphine, isothiocyanates, isocyanates, acyl azides, N-hydroxyl esters, sulfonyl chlorides, aldehydes, glyoxals, epoxides, oxiranes, carbonates, aryls, carbodiimides, and carboxylic acid anhydrides.

[0114] Carbodiimides, diazoalkanes, diazoacetyl compounds, carbonyldiimidazoles are reactive groups that can react with carboxyls and amines.

[0115] Reactive groups that can react with thiols include maleimide, haloacetamide, pyridyl disulfide, thiosulfone, vinyl sulfone, haloacetyl, aziridine, acryloyl, and aryl.

[0116] Reactive groups that can react with aldehydes include hydrazide and alkoxyamine, and reactive groups that can react with hydroxyl include epoxy, oxirane, carbonyldiimidazole, N,N'-disuccinimidyl carbonate, N-hydroxysuccinimidyl chloroformate, and isocyanate.

[0117] Isocyanate is a reactive group that can react to hydroxyl.

[0118] Photoreactive reactive groups include diazirine, aryl azide, aryl, benzophenol, and diazo compounds.

[0119] Specific examples of the linker having the reactive group include the following:

[0120] Examples of linkers having the same reactive group terminal include linkers having N-hydroxysuccinimide ester as the 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・2HCl (DMA), Dimethyl pimelimidate・2HCl (DMP), Dimethyl suberimidate・2HCl (DMS), Dimethyl3,3'-dithiobispropionimidate・2H Cl (DTBP), 1,5-difluoro-2,4-dinitrobenzene (DFDNB), Disuccinimidyl tartrate (DST), Bis[2-(Succinimidooxycarbonyloxy)ethyl]Sulfone (BSOCOES), and linkers with maleimide as a reactive group (e.g., Bismaleimidoethane (BMOE), 1,4-bismaleimidobutane (BMB), Bismaleimidohexane (BMH), Tris(2-maleimidothyl)amine (TMEA), 1,8-bismaleimido-(PEG) ２ (BM (PEG) ２ ), 1,8-bismaleimido-(PEG) ３ (BM (PEG) ３ ), Dithiobismaleimidoethane (DTME) is used.

[0121] The main linkers having different reactive group ends include linkers having NHS ester and maleimide 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) １２ , SM(PEG) ２４ ), a linker having an NHS ester and a pyridyldithiol as a reactive group (e.g., SPDP, LC-SPDP, Sulfo-LC-SPDP, SMPT, (PEG) ４ SPDP, PEG12-SPDP), linkers with NHS ester and haloacetyl as reactive groups (e.g., SIA, SBAP, SIAB, Sulfo-SIAB), linkers with NHS ester and aryl azide as reactive groups (e.g., ANB-NOS, Sulfo-SANPAH, ATFB), linkers with NHS ester and diazirine as reactive groups (e.g., SDA, Sulfo-SDA, LC-SDA, SDAD, Sulfo-SDAD), carbodiimide Linkers having a hydrazide as a reactive group (e.g., DCC, EDC, EDAC, NHS, Sulfo-NHS), linkers having a maleimide and a hydrazide as a reactive group (e.g., BMPH, EMCH, MPBH, KMUH), linkers having a pyridyldithiol and a hydrazide as a reactive group (e.g., PDPH), linkers having an isocyanate and a maleimide as a reactive group (e.g., PMPI), and linkers having an NHS ester and a psoralen as a reactive group (e.g., SPB) are used.

[0122] Other linkers include polypeptide-containing linkers, such as Fmoc-Ala-Ala-Asn-PAB, Fmoc-Ala-Ala-Asn(Trt)-PAB, and Fmoc-PEG. ３ -Ala-Ala-Asn(Trt)-PAB, Fmoc-PEG ４ -Ala-Ala-Asn(Trt)-PAB, Fmoc-Ala-Ala-Asn-PAB-PNP, Fmoc-Ala-Ala-Asn(Trt)-PAB-PNP, Fmoc-PEG ３ -Ala-Ala-Asn(Trt)-PAB-PNP, Azide-PEG ４ -Ala-Ala-Asn(Trt)-PAB-PNP, Mal-PEG ４-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. are used.

[0123] Also, Bis-PEG-acid, PEG Acid (e.g., Acid-PEG-TEMPO, Amino-PEG-acid, Amino-PEG-CH ２ COOH, Aminoxy-PEG-acid, Azido-PEG-acid, Carboxy-PEG-sulfonic acid, Fmoc-N-amido-PEG-acid, Fmoc-N-amido-PEG-CH ２ COOH, Fmoc-aminoxy-PEG-acid, Hydroxy-PEG-acid, Hydroxy-PEG-CH ２ CO ２ H, m-PEG-acid, m-PEG-(CH ２ ) ３ -acid, Methoxytrityl-N-PEG-acid, N-methyl-N-(t-Boc)-PEG-acid, Propargyl-PEG-acid, Propargyl-PEG-CH ２ COOH, Propargyl-PEG-(CH ２ ) ３ -acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG-CH ２COOH, t-Boc-Aminoxy-PEG-acid, Acid-PEG-PFP ester, Miscellaneous PEG acid, ), PEG PFP ester (e.g., Acid-PEG-PFP ester, Bis-PEG-PFP ester), Bis-PEG-NHS, PEG Aldehyde (e.g., m-PEG-aldehyde, m-PEG-benzaldehyde, Ald-PEG-acid, Ald-PEG-amine, Ald-PEG-azide, Ald-PEG-NH-Boc, Ald-PEG-NHS ester, Ald-PEG-TFP ester, Ald-PEG-t-butyl ester), PEG Tosylate (e.g., Azido-PEG-Tos, Hydroxy-PEG-Tos, m-PEG-Tos, t-Boc-Aminoxy-PEG-Tos, Trifluoroethyl-PEG-Tos, Tos-PEG-acid, Tos-PEG-CH ２ COOH, Tos-PEG-alkyne, Tos-PEG-t-butyl ester, Tos-PEG-CH ２ CO ２ tBu, Tos-PEG-Tos, S-acetyl-PEG ６ -Tos, N-Tos-N-(t-butoxycarbonyl)-aminoxy-PEG ４ -Tos, Ms-PEG-Ms, Ms-PEG-t-butyl ester, PEG-Ms, Propargyl-PEG-Ms), Boc-PEG (e.g., Amino-PEG-t-Boc-Hydrazide, Azido-PEG-t-Boc-Hydrazide, Boc-NH-PEG-NH-Boc, Bromoacetamido-PEG-Boc-amine, m-PEG-ONHBoc, Mal-Alkyl-t-Boc-amine, N-Boc-PEG-alcohol, N-Boc-PEG-bromide, N-methyl-N-(t-Boc)-PEG-acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG-CH ２COOH, 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), PEG NHS 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-PEG-NHS ester), Fmoc-PEG (e.g., Fmoc-N-amido-PEG-acid, Fmoc-NH-PEG-CH ２ COOH, Fmoc-PEG-NHS ester), Biotin PEG (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-PEG ３ -SS-azide, DBCO-S-S-PEG ３ -Biotin, Dde Biotin-PEG ４ -Alkyne, Dde Biotin-PEG ４ -Azide, Dde Biotin-PEG ４ -DBCO, Diazo Biotin-PEG ３ -Alkyne, Diazo Biotin-PEG ３ -Azide, Diazo Biotin-PEG ３-DBCO、Diol Biotin-PEG ３ -Alkyne、Diol Biotin-PEG ３ -Azide、PC Biotin-PEG ３ -Alkyne、PC-Biotin-PEG ４ -PEG ４ -Alkyne、PC-Biotin-PEG ４ -PEG4-Alkyne、PC Biotin-PEG ３ -Azide、PC-Biotin-PEG ４ -PEG ３ -Azide、PC-Biotin-PEG ４ -NHS carbonate、PC DBCO-PEG ３ -Biotin、WSPC Biotin-PEG ３-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-CH ２ COOH, Propargyl-PEG-(CH ２ ) ３ -acid, Propargyl-PEG-(CH ２ ) ３-methyl ester、Prropargyl-･Gmcrrylat e、Propargglll･･･alcohol、rrop argylmm･･mamine、.. G-methylamiine、| Propargylla. de、Propargyllm･･bromide、rro pargyl-･･omMaleimide、rropa gylmPEy-ｭs、Propargyllmm･om｢ｳester、PrropargylllEGsulffoiic I ester、Propargyl-･ym｣ｨ ２ 39 ２ tBu、Propargylll･･thiol、ｰcbd gummies carbonaate、 nooxy-EEorropargyl、 BismPropargyl-･1、mm PEGPropargyll) . ２ 3OH、Azido-EE(((3(H ２ ) ) ３ -methyl ester、｡zidomE･m｡crylatee｡ do-PEGalcoholl ２ ) ) ３ OH、｡zido-EGamine、｡zid o-PEmazide、｡zido-EMM I ylamine、｡zido-･mmethyll ester、｡zido-EE1mｮｳ esterr ２ 39 ２-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-CH ２ CO ２ -t-butyl ester, Azido-PEG-TFP ester, Azido-PEG-Tos, Aminoxy-PEG-azide, Bromo-PEG-azide, Bromoacetamide-PEG-azide, Carboxyrhodamine 110-PEG-Azide, Isothiocyanato-PEG-Azide, Isothiocyanato- PEG-Azide, m-PEG-azide, Propargyl-PEG-azide, TAMRA-PEG-Az ide, t-Boc-N-Amido-PEG-Azide, t-Boc-Aminooxy-PEG-Azide, T hiol-PEG-Azide, Trifluoroethyl-PEG-Azide, Azido-PEG-amino acid, Azido-PEG ４ -4-nitrophenyl carbonate, S-acetyl-PEG ３ -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 is used.

[0124] In another embodiment, polyethylene glycol (PEG) as described in WO2015 / 057699 and WO2017 / 165851 can be used to obtain a conjugate that is expected to have better in vivo kinetics.

[0125] The linker mediated between the anti-CAPRIN-1 antibody and the topoisomerase I inhibitor may be composed of a single type or multiple types.

[0126] Methods for preparing a conjugate of an anti-CAPRIN-1 antibody and a topoisomerase I inhibitor include a method in which a topoisomerase I inhibitor or a derivative thereof is conjugated using the ε-amino group of a lysine side chain of the antibody, and a method in which a cysteine ​​residue forming a disulfide bond of the antibody is conjugated using a thiol formed by reduction treatment. Reduction treatment methods and methods for drug conjugation after reduction include those described in WO2019 / 191630, WO2015 / 031698, WO2017 / 201204, WO2015 / 031541, and WO2015 / 031693.

[0127] When the ε-amino group of a lysine residue of an antibody is used, for example, an amide bond is formed by reacting it with an active ester (e.g., N-hydroxysuccinimide ester). In this case, since there are many lysine residues in an antibody, the binding reaction proceeds nonspecifically.

[0128] When using a thiol that forms a disulfide bond in the side chain of a cysteine ​​residue on an antibody, a method is used in which the disulfide bond on the antibody is converted to a thiol using a reducing agent such as mercaptoethanol, followed by reaction with a maleimide or α-haloamide. Furthermore, methods using sulfonephenyloxadiazole, 4-cyanoethynyloxy derivatives, etc., can be used to stabilize thiol-mediated bonds. These bonds are more stable for longer periods than bonds formed by the conjugation reaction of cysteine ​​to maleimide. Furthermore, since stability is improved when the imide ring formed by the thiol group attached to the maleimide is opened by hydrolysis to form an amide bond, a linker with an amino group near the imide group can also be used. Furthermore, the thiol of the cysteine ​​residue on an antibody forms a disulfide bond, and a topoisomerase I inhibitor can be attached between the two thiols. For example, a cross-linked bond can be formed using a linker with two disulfide bond sites, which can be generated from an amide group with two sulfones at the β-position, or dibromomaleimide.

[0129] The conjugates of the present invention can be produced, for example, using THIOMAB™ technology, which is a method for introducing a fixed number of thiol groups into a specific portion of an antibody (see Nature Biotechnology 26, 925-932 (2008)), or using ThioBridge™ technology (see Methods Mol Biol. 2078: 113-129 (2020)), or RESPECT™ technology (see MAbs. 9 (6): 907-915 (2017)).

[0130] The conjugate of the present invention can be prepared, for example, by reducing an antibody with the reducing agent dithiothreitol (DTT) in a phosphate buffer solution to obtain an antibody having a thiol reactive group, and then forming a conjugate with a topoisomerase I inhibitor or a derivative thereof. In addition to the method using a reducing agent, the conjugate can also be obtained by adding a thiol group to the primary amine of a lysine residue in the antibody by introducing Traut's reagent (2-Iminothiolane or N-Succinimidyl S-Acetylthioacetate (SATA)).

[0131] The amount of thiol added 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 phosphate buffer (pH 8.0) and distilled water, adding a DTNB solution dissolved in phosphate buffer, Good's buffer, or Tris buffer, incubating for a certain period of time, and then measuring the absorbance at 412 nm (see G. L. Ellman, Arch. Biochem. Biophys., 82, 70 (1959)).

[0132] The thiol groups added by cleaving disulfide bonds in the antibody through reduction treatment are preferably subjected to a treatment (capping) to prevent re-formation of disulfide bonds, such as with N-ethylmaleimide (NEM) or 2-iodoacetamide (IAA).

[0133] Forming a conjugate by binding a topoisomerase I inhibitor using a thiol group added to an antibody can be carried out by known methods. Specifically, for example, linker reagents having a maleimide group or a bromoacetamide group can be used as linker reagents that specifically bind to the thiol group of a 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, if an amino group is present on the drug side, a conjugate can be obtained by forming an amide bond with the N-succinimide group of SMCC.

[0134] In another embodiment, an amide bond is first formed with SMCC at the amino group present on the drug side, and then the thiol group added to the antibody side is reacted with the maleimide group of SMCC on the drug side to obtain a conjugate.

[0135] In another embodiment, a conjugate can be formed using two linkers. For example, a conjugate can be prepared by forming an amide bond between a primary amino group present in a lysine residue on the antibody and the N-succinimide group of SATA (N-succinimidyl-S-acetylthioacetate), adding a thiol group to the antibody, and then synthesizing a conjugate having an amino group on the drug side, or a conjugate having an amino group added thereto according to a standard method, by reacting SMCC to form an amide bond with the N-succinimide group on the SMCC, and then a conjugate can be obtained by reacting the maleimide group on the SMCC on the drug side with the thiol group on the SATA bound to the antibody.

[0136] In another embodiment, conjugates of an antibody and a topoisomerase I inhibitor or a derivative thereof can be prepared by, for example, using maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-Val-Cit-PAB) as a linker. mc-val-Cit-PAB (mc-vc-PAB) is a linker that can be cleaved by intracellular proteases (e.g., cathepsin B). A thiol group is attached to an antibody dissolved in phosphate buffer using DTT or the like. On the other hand, a topoisomerase I inhibitor or a derivative thereof having an amino group is reacted with the benzyloxycarbonyl (PAB) in mc-Val-Cit-PAB to prepare a topoisomerase I inhibitor or a derivative thereof bound to mc-val-Cit-PAB, and this can then be reacted with the aforementioned antibody to which the thiol has been attached to obtain a conjugate.

[0137] In yet another embodiment, SATA is attached to a primary amino group of a lysine residue of an antibody to add a thiol group, while succinimidyl 3-(2-pyridyldithio)propionate (SPDP) is reacted with an amino group on the drug to form an amide bond with the N-succinimide group of SPDP.

[0138] The linker used in the present invention can be one that is cleavable under intracellular conditions. When such a linker is used, a substance having antitumor activity, comprising a topoisomerase I inhibitor or a topoisomerase I inhibitor and a portion of the linker, is liberated intracellularly. For example, the linker is cleaved by intracellular peptidases or proteases. Preferred linkers are those that are cleaved by lysosomal or endosomal proteases, cathepsin B, cathepsin D, or plasmin. Examples include linkers containing polypeptides (Val-Cit, Phe-Leu, or Gly-Phe-Leu-Gly) that can be cleaved by cathepsin B. More specifically, the linkers described in U.S. Patent No. 6,214,345 can be used.

[0139] Furthermore, in another embodiment, as a means for improving the stability, solubility, and metabolic property of the conjugate of the present invention in blood and the binding affinity of the topoisomerase I inhibitor or its derivative to an anti-CAPRIN-1 antibody, for example, a linker having glucuronic acid (preferably β-D-glucuronide) described in WO 2007 / 011968 can be used. Alternatively, means described in WO 2013 / 173337, WO 2015 / 095755, WO 2015 / 123679, and WO 2018 / 031690 can be used.

[0140] Furthermore, in another embodiment, the topoisomerase I inhibitor or a derivative thereof can be site-specifically bound to an anti-CAPRIN-1 antibody using, for example, the means described in WO2006 / 65533 and WO2018 / 160683.

[0141] In yet another embodiment, a conjugate of an anti-CAPRIN-1 antibody to which two or more drugs, including a topoisomerase I inhibitor, are bound can be obtained by the method described in WO2018 / 112253.

[0142] To obtain a composition containing the conjugate of the present invention, for example, the antibody can be subjected to gel filtration chromatography or the like, and a peak with a higher molecular weight than that of the antibody before the linker is attached can be isolated. To detect the mass of the conjugate while maintaining the intact bivalent antibody, for example, the method described in WO2013 / 049410 can be used.

[0143] The number of topoisomerase I inhibitors bound per antibody molecule of the conjugate of the present invention can be quantified according to known methods such as mass spectrometry, ELISA, electrophoresis, and chromatography such as HPLC.

[0144] <Anti-tumor effect of conjugate> The conjugate of the present invention has anti-tumor activity in vitro or in vivo. Anti-tumor activity means reduction, elimination, inhibition of growth, apoptosis, necrosis, or killing of targeted cancer cells. Therefore, the anti-tumor effect of the conjugate of the present invention can be determined by examining the anti-tumor activity against cancer.

[0145] The in vivo antitumor effect can be assessed by administering the conjugate to a cancer-bearing organism, measuring the size of the tumor after administration, and examining the size of the cancer over time. The antitumor effect of the present invention can also be assessed by examining the survival rate. It can also be assessed by examining the ability to produce cytokines or chemokines. The antitumor effect of the conjugate of the present invention can be further assessed by examining the prevention of cancer, metastasis, or recurrence.

[0146] The conjugate of the present invention is expected to have a stronger antitumor effect if it has a higher binding affinity with the CAPRIN-1 protein on the surface of cancer cells. 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 It is desirable that:

[0147] The ability of the conjugate of the present invention to bind to CAPRIN-1 can be determined using binding assays such as surface plasmon resonance (SPR), ELISA, Western blotting, immunofluorescence, and flow cytometry.

[0148] As described above, the conjugates of the present invention have an enhanced anti-tumor effect compared to an anti-CAPRIN-1 antibody alone, and the enhancement rate is preferably 30% or more, more preferably 40% or more, even more preferably 50% or more, even more preferably 55% or more, even more preferably 60% or more, even more preferably 65% ​​or more, and most preferably 70% or more. The enhancement rate of the anti-tumor effect of the conjugates of the present invention compared to an anti-CAPRIN-1 antibody alone can be calculated by administering an effective amount of each to tumor-bearing mice under the same conditions and comparing the tumor volumes on or after day 10 after the start of administration.

[0149] <Pharmaceutical Composition, Method for Treating and / or Preventing Cancer> 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) that expresses the CAPRIN-1 protein.

[0150] As used herein, the terms "tumor" and "cancer" refer to malignant neoplasms and are used interchangeably.

[0151] The cancers targeted by the present invention may be any cancers that express CAPRIN-1 protein on the cell membrane surface. Preferred cancers include breast cancer, kidney cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cell tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, and multiple myeloma, as well as testicular cancer, thyroid cancer, head and neck cancer, and urothelial cancer.

[0152] More specifically, the cancers include, for example, breast adenocarcinoma, hybrid breast adenocarcinoma, malignant mixed breast tumor, intraductal papillary adenocarcinoma, recurrent metastatic breast cancer, lung adenocarcinoma, non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, neuroepithelial tissue tumors such as glioma, glioblastoma, neuroblastoma, ependymoma, neuronal tumor, embryonal neuroectodermal tumor, schwannoma, neurofibroma, meningioma, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, digestive lymphoma, small to medium cell lymphoma, cecum 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 ductal carcinoma, invasive pancreatic ductal carcinoma, adenocarcinoma of pancreatic cancer, metastatic adenocarcinoma, Acinar cell carcinoma, adenosquamous carcinoma, giant cell tumor, intraductal papillary mucinous neoplasm, mucinous cystadenocarcinoma, pancreatoblastoma, pancreatic head cell tumor, Frants' tumor, serous cystadenocarcinoma, solid papillary carcinoma, gastrinoma, glucagonoma, insulinoma, multiple endocrine neoplasia 1 (Wermer's syndrome), nonfunctioning islet cell tumor, somatostatinoma, VIP-secreting tumor, cervical cancer, endometrial cancer, fibrosarcoma, bone and joint sarcoma, Ewing's sarcoma, Wilms' tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, malignant soft tissue tumor, teratoma group tumor, head and neck cancer includes, but is not limited to, hypopharyngeal cancer, oropharynx cancer, tongue cancer, nasopharyngeal cancer, oral cancer, lip cancer, paranasal sinus cancer, laryngeal cancer, recurrent brain tumor, etc.

[0153] Furthermore, preferred subjects (patients) are mammals, including, for example, primates, pet animals, livestock, sport animals, etc., with humans, dogs, and cats being particularly preferred.

[0154] When the conjugates used in the present invention are used as pharmaceutical compositions, they can be formulated by methods known to those skilled in the art. For example, they can be used parenterally in the form of a sterile solution or suspension injection in water or other pharmaceutically acceptable liquid. For example, they can be formulated by appropriately combining them with pharmacologically acceptable carriers or vehicles, specifically, sterilized water, physiological saline, vegetable oils, emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, excipients, binders, etc., and mixing them in a unit dosage form required for generally accepted pharmaceutical practice. The amount of active ingredient in these formulations is such that an appropriate dose within the indicated range can be obtained.

[0155] When the conjugate of the present invention is used as a pharmaceutical composition, it can be formulated in a lyophilized state containing any salt, surfactant, buffer, sugar, and cryoprotectant (including some sugars). For example, a pharmaceutical composition solution described in WO2007 / 019232 containing excipients including sucrose, polysorbate 20, polysorbate 80, cyclodextrin, glucose, glycerol, polyethylene glycol, mannitol, sodium chloride, and an amino acid, and having a pH of 4 to 8, preferably 4.5 to 7.6, can be used.

[0156] Sterile compositions for injection can be formulated according to standard pharmaceutical practice using a vehicle such as distilled water for injection. Aqueous solutions for injection may be formulated with, for example, physiological saline, an isotonic solution containing D-sorbitol, D-mannose, D-mannitol, or other auxiliary agents, alcohol (specifically, ethanol), polyalcohol (specifically, propylene glycol, polyethylene glycol), nonionic surfactants (specifically, polysorbate 80™, HCO-60), solubilizers such as benzyl benzoate and benzyl alcohol, oily solutions (specifically, sesame oil, soybean oil), buffers (specifically, phosphate buffer, sodium acetate buffer), soothing agents (specifically, procaine hydrochloride), stabilizers (specifically, benzyl alcohol, phenol), and antioxidants. The prepared injection solutions are usually filled into appropriate ampoules.

[0157] Administration may be oral or parenteral, preferably parenteral, and specific examples include injections, intranasal administrations, pulmonary administrations, transdermal administrations, etc. Examples of injections include intravenous injections, intramuscular injections, intraperitoneal injections, subcutaneous injections, intratumoral injections, etc., which can be used for systemic or local administration.

[0158] Furthermore, an appropriate administration method can be selected depending on the patient's age, body weight, sex, symptoms, etc. The dosage of a pharmaceutical composition containing an antibody or a polynucleotide encoding an antibody can be selected, for example, from the range of 0.0001 mg to 1000 mg per kg of body weight per administration, such as 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 per administration. Alternatively, the dosage can be selected, for example, from the range of 0.001 to 100,000 mg / body per patient, although it is not necessarily limited to these values.

[0159] The dosage and administration method will vary depending on the patient's weight, age, sex, symptoms, etc., but can be appropriately selected by those skilled in the art.

[0160] By administering to a subject a pharmaceutical composition for treating and / or preventing cancer, which contains the conjugate of the present invention as an active ingredient, cancers that express CAPRIN-1 on the cell membrane surface, preferably breast cancer, kidney cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder 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, and urothelial cancer, can be treated and / or prevented.

[0161] The present invention will be specifically described below based on examples, but the scope of the present invention is not limited to these specific examples.

[0162] Example 1: Anti-CAPRIN-1 Polyclonal Antibody The anti-CAPRIN-1 polyclonal antibody immunologically reactive with the CAPRIN-1 protein used in the conjugate of the present invention was prepared by mixing 1 mg of human CAPRIN-1 recombinant protein represented by SEQ ID NO: 2 and SEQ ID NO: 4, prepared according to Example 3 of WO 2010 / 016526, with an equal volume of incomplete Freund's adjuvant (IFA) solution. This mixture was subcutaneously administered to rabbits four times every two weeks. Blood was then collected to obtain antisera containing the polyclonal antibody. The obtained antisera was 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). Additionally, serum from a rabbit not administered the antigen was purified using a protein G carrier in the same manner as above to serve as a rabbit control antibody.

[0163] Furthermore, the following polyclonal antibodies #2 to #6 against partial polypeptides of the CAPRIN-1 protein were obtained in the same manner as in the preparation of the polyclonal antibodies against the CAPRIN-1 protein.

[0164] Anti-CAPRIN-1 polyclonal antibody #2 against the partial polypeptide represented by SEQ ID NO: 37 (SEQ ID NO: 31 herein) described in WO2011 / 096528.

[0165] Anti-CAPRIN-1 polyclonal antibody #3 against the partial polypeptide represented by SEQ ID NO: 5 (SEQ ID NO: 32 herein) described in WO2013 / 018894.

[0166] Anti-CAPRIN-1 polyclonal antibody #4 against the partial polypeptide represented by SEQ ID NO: 5 (SEQ ID NO: 33 herein) described in WO2013 / 125654.

[0167] Anti-CAPRIN-1 polyclonal antibody #5 against the partial polypeptide represented by SEQ ID NO: 37 (SEQ ID NO: 34 herein) described in WO2011 / 096533.

[0168] Anti-CAPRIN-1 polyclonal antibody #6 against a partial polypeptide represented by SEQ ID NO: 37 (SEQ ID NO: 35 herein) described in WO2011 / 096534.

[0169] Example 2 Anti-CAPRIN-1 Monoclonal Antibody The following anti-CAPRIN-1 monoclonal antibody was used in the conjugate of the present invention.

[0170] An anti-CAPRIN-1 monoclonal antibody described in WO2011 / 096528, comprising the amino acid sequence of a heavy chain variable region represented by SEQ ID NO:39, and the amino acid sequence of a light chain variable region represented by SEQ ID NO:43, wherein CDR1 to CDR3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO:36, SEQ ID NO:37, and SEQ ID NO:38, respectively, and CDR1 to CDR3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42, respectively.

[0171] An anti-CAPRIN-1 monoclonal antibody described in WO2015 / 020212, comprising the amino acid sequence of a heavy chain variable region represented by SEQ ID NO:47 and the amino acid sequence of a light chain variable region represented by SEQ ID NO:51, wherein CDR1 to CDR3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO:44, SEQ ID NO:45, and SEQ ID NO:46, respectively, and CDR1 to CDR3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:50, respectively.

[0172] An anti-CAPRIN-1 monoclonal antibody described in WO2011 / 096519, comprising the amino acid sequence of a heavy chain variable region represented by SEQ ID NO:55 and the amino acid sequence of a light chain variable region represented by SEQ ID NO:59, wherein CDR1 to CDR3 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 CDR3 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.

[0173] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 125654, comprising the amino acid sequence of a heavy chain variable region represented by SEQ ID NO: 63 and the amino acid sequence of a light chain variable region represented by SEQ ID NO: 67, wherein CDR1 to CDR3 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 CDR3 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.

[0174] An anti-CAPRIN-1 monoclonal antibody described in WO2011 / 096517, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 68 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 69.

[0175] Anti-CAPRIN-1 monoclonal antibodies are disclosed in WO2011 / 096528, including an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 70 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 71; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 72 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 73; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 75; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 76 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 77; and an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 78 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 79.

[0176] An anti-CAPRIN-1 monoclonal antibody described in WO2011 / 096533, comprising an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 80 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 81; and an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 82 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 83.

[0177] An anti-CAPRIN-1 monoclonal antibody described in WO2011 / 096534, comprising an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 84 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 85; and an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 86 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 87.

[0178] Anti-CAPRIN-1 monoclonal antibodies described in WO2010 / 016526 include an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 88 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 89; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 90 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 91; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 92 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 93; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 94; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 96 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 97; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 98 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 99; and an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 100 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 101.

[0179] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 018894, comprising an antibody having a heavy chain variable region with the amino acid sequence of SEQ ID NO: 102 and a light chain variable region with the amino acid sequence of SEQ ID NO: 103; and an antibody having a heavy chain variable region with the amino acid sequence of SEQ ID NO: 104 and a light chain variable region with the amino acid sequence of SEQ ID NO: 105.

[0180] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 018892, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 106 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 107.

[0181] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 018891, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 108 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 109.

[0182] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 018889, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 111.

[0183] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 018883, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 113.

[0184] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 125636, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 115.

[0185] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 125654, comprising an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 116 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 117; and an antibody having a heavy chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 118 and a light chain variable region with an amino acid sequence represented by the amino acid sequence of SEQ ID NO: 119.

[0186] An anti-CAPRIN-1 monoclonal antibody described in WO2013 / 125630, comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 121.

[0187] an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 122 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 123; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 124 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 125; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 127; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 128 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 129; an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 131; and an antibody comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 132 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 133.

[0188] For the above-mentioned anti-CAPRIN-1 monoclonal antibodies, a nucleotide sequence was designed so that a heavy chain variable region in which CDR1 to CDR3 consist of the amino acid sequences of SEQ ID NOs: 36, 37, and 38, respectively, and in which the framework regions comprise a human antibody sequence, could be expressed, and inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted. Similarly, a nucleotide sequence was designed so that a light chain variable region in which CDR1 to CDR3 consist of SEQ ID NOs: 40, 41, and 42, respectively, and in which the framework regions comprise a human antibody sequence, could be expressed, and 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 a standard method to obtain a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #1 (humanized antibody #1), in which CDRs 1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NOs: 36, 37, and 38, respectively, and CDRs 1 to 3 of the light chain variable region consist of the amino acid sequences of SEQ ID NOs: 40, 41, and 42, respectively.

[0189] Similarly, a nucleotide sequence was designed to enable expression of a heavy chain variable region represented by SEQ ID NO: 47, in which CDRs 1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 46, respectively, and in which the framework regions comprise the sequence of a human antibody, and this was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted. Similarly, a base sequence was designed to enable expression of a heavy chain variable region represented by SEQ ID NO:51, in which CDR1 to 3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:50, respectively, and the framework regions comprise the sequence of a human antibody. This base sequence was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted, and the two recombinant expression vectors were introduced into mammalian cells according to standard methods to obtain a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #2 (humanized antibody #2), in which CDR1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO:44, SEQ ID NO:45, and SEQ ID NO:46, and CDR1 to 3 of the light chain variable region consist of the amino acid sequences of SEQ ID NO:48, SEQ ID NO:49, and SEQ ID NO:50, respectively.

[0190] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #3 (humanized antibody #3) was obtained, in which CDRs 1 to 3 of the heavy chain variable region consisted of the amino acid sequences of SEQ ID NOs: 52, 53, and 54, respectively, and CDRs 1 to 3 of the light chain variable region consisted of the amino acid sequences of SEQ ID NOs: 56, 57, and 58, respectively.

[0191] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #4 (humanized antibody #4) was obtained, in which CDRs 1 to 3 of the heavy chain variable region consisted of the amino acid sequences of SEQ ID NOs: 60, 61, and 62, respectively, and CDRs 1 to 3 of the light chain variable region consisted of the amino acid sequences of SEQ ID NOs: 64, 65, and 66, respectively.

[0192] Similarly, culture supernatants containing the following humanized anti-CAPRIN-1 monoclonal antibodies #9 to #41 (humanized antibodies #9 to #41) were obtained.

[0193] Humanized anti-CAPRIN-1 monoclonal antibody #9 (humanized antibody #9) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 68 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 69.

[0194] Humanized anti-CAPRIN-1 monoclonal antibody #10 (humanized antibody #10) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 70 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 71.

[0195] Humanized anti-CAPRIN-1 monoclonal antibody #11 (humanized antibody #11) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 72 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 73.

[0196] Humanized anti-CAPRIN-1 monoclonal antibody #12 (humanized antibody #12) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 75.

[0197] Humanized anti-CAPRIN-1 monoclonal antibody #13 (humanized antibody #13) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 76 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 77.

[0198] Humanized anti-CAPRIN-1 monoclonal antibody #14 (humanized antibody #14) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 78 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 79.

[0199] Humanized anti-CAPRIN-1 monoclonal antibody #15 (humanized antibody #15) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 80 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 81.

[0200] Humanized anti-CAPRIN-1 monoclonal antibody #16 (humanized antibody #16) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 82 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 83.

[0201] Humanized monoclonal antibody #17 (humanized antibody #17) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 84 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 85.

[0202] Humanized anti-CAPRIN-1 monoclonal antibody #18 (humanized antibody #18) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 86 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 87.

[0203] Humanized anti-CAPRIN-1 monoclonal antibody #19 (humanized antibody #19) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 88 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 89.

[0204] Humanized anti-CAPRIN-1 monoclonal antibody #20 (humanized antibody #20) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 90 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 91.

[0205] Humanized anti-CAPRIN-1 monoclonal antibody #21 (humanized antibody #21) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 92 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 93.

[0206] Humanized anti-CAPRIN-1 monoclonal antibody #22 (humanized antibody #22) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 94 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 95.

[0207] Humanized anti-CAPRIN-1 monoclonal antibody #23 (humanized antibody #23) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 96 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 97.

[0208] Humanized anti-CAPRIN-1 monoclonal antibody #24 (humanized antibody #24) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 98 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 99.

[0209] Humanized anti-CAPRIN-1 monoclonal antibody #25 (humanized antibody #25) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 100 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 101.

[0210] Humanized anti-CAPRIN-1 monoclonal antibody #26 (humanized antibody #26) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 102 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 103.

[0211] Humanized anti-CAPRIN-1 monoclonal antibody #27 (humanized antibody #27) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 104 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 105.

[0212] Humanized anti-CAPRIN-1 monoclonal antibody #28 (humanized antibody #28) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 106 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 107.

[0213] Humanized anti-CAPRIN-1 monoclonal antibody #29 (humanized antibody #29) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 108 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 109.

[0214] Humanized anti-CAPRIN-1 monoclonal antibody #30 (humanized antibody #30) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 111.

[0215] Humanized anti-CAPRIN-1 monoclonal antibody #31 (humanized antibody #31) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 113.

[0216] Humanized anti-CAPRIN-1 monoclonal antibody #32 (humanized antibody #32) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 115.

[0217] Humanized anti-CAPRIN-1 monoclonal antibody #33 (humanized antibody #33) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 117.

[0218] Humanized anti-CAPRIN-1 monoclonal antibody #34 (humanized antibody #34) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 118 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 119.

[0219] Humanized anti-CAPRIN-1 monoclonal antibody #35 (humanized antibody #35) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 121.

[0220] Humanized anti-CAPRIN-1 monoclonal antibody #36 (humanized antibody #36) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 122 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 123.

[0221] Humanized anti-CAPRIN-1 monoclonal antibody #37 (humanized antibody #37) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 124 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 125.

[0222] Humanized anti-CAPRIN-1 monoclonal antibody #38 (humanized antibody #38) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 127.

[0223] Humanized anti-CAPRIN-1 monoclonal antibody #39 (humanized antibody #39) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 128 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 129.

[0224] Humanized monoclonal antibody #40 (humanized antibody #40) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 131.

[0225] Humanized anti-CAPRIN-1 monoclonal antibody #41 (humanized antibody #41) comprising the amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 132 and the amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 133.

[0226] Furthermore, for humanized antibody #1 of the above anti-CAPRIN-1 monoclonal antibodies, a nucleotide sequence was designed to enable expression of a heavy chain variable region in which CDRs 1 to 3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NOs: 36, 37, and 38, respectively, and in which the framework regions comprise the sequence of a human antibody, and this was inserted into a mammalian expression vector containing the heavy chain constant region of human IgG1 in which serine (Ser), amino acid 239 (EU numbering), has been substituted with aspartic acid (Asp) and isoleucine (Ile), amino acid 332 (EU numbering), has been substituted with glutamic acid (Glu). Furthermore, a nucleotide sequence was designed to enable expression of a light chain variable region in which CDRs 1 to 3 of the light chain variable region consist of SEQ ID NOs: 40, 41, and 42, respectively, and in which the framework regions comprise the sequence of a human antibody, and this was inserted into a mammalian expression vector containing the light chain constant region of human IgG1. The two recombinant expression vectors were introduced into mammalian cells according to standard methods to obtain a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #5 (humanized antibody #5), which consisted of the heavy chain full-length amino acid sequence consisting of the heavy chain variable region prepared above and a heavy chain constant region of human IgG1 in which serine (Ser), amino acid 239 (EU numbering), has been substituted with aspartic acid (Asp) and isoleucine (Ile), amino acid 332 (EU numbering), has been substituted with glutamic acid (Glu), and the light chain full-length amino acid sequence consisting of the light chain variable region and human light chain constant region prepared above.

[0227] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #6 (humanized antibody #6) consisting of 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.

[0228] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #7 (humanized antibody #7) consisting of 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.

[0229] Similarly, a culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #8 (humanized antibody #8) consisting of 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.

[0230] Similarly, culture supernatants containing humanized anti-CAPRIN-1 antibodies #42 to #74 (humanized antibodies #42 to #74) each consisting of the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region of each of the humanized antibodies #9 to #41 prepared above were obtained.

[0231] The culture supernatants containing the obtained humanized anti-CAPRIN-1 monoclonal antibodies #1 to #74 were purified using Hitrap Protein A Sepharose FF (GE Healthcare) according to standard methods, and then substituted with PBS(-) and filtered through a 0.22 μm filter (Millipore) to prepare samples.

[0232] (Example 3) Preparation of conjugate of anti-CAPRIN-1 antibody and topoisomerase I inhibitor (SN38) As a conjugate of anti-CAPRIN-1 antibody and topoisomerase I inhibitor, a conjugate of anti-CAPRIN-1 and SN38 (SN38 conjugate) was prepared according to a standard method.

[0233] Conjugates of anti-CAPRIN-1 polyclonal antibodies #1 to #6 described in Example 1 with SN38 were prepared according to standard methods. Purified anti-CAPRIN-1 polyclonal antibody #1 was dissolved in PBS(-) at a concentration of 10 mg / ml. Anti-CAPRIN-1 polyclonal antibody #1 was reduced to a solution containing the polyclonal antibody #1 using TCEP (TRIS(2-carboxyethyl)phosphine) according to standard methods, and then SN38, a topoisomerase I inhibitor, was conjugated. Specifically, a compound CL2A-SN38 (CAS No. 1279680-68-0) in which a linker is attached to SN38 was used and conjugated to anti-CAPRIN-1 polyclonal antibody #1 according to a standard method to obtain a solution containing the anti-CAPRIN-1 polyclonal antibody #1-CL2A-SN38 of the present invention (SN38 conjugate 1).

[0234] Similarly, solutions containing conjugates using CL2A-SN38 were obtained for anti-CAPRIN-1 polyclonal antibodies #2 to #6 (the conjugate using anti-CAPRIN-1 polyclonal antibody #2: SN38 conjugate 2, the conjugate using anti-CAPRIN-1 polyclonal antibody #3: SN38 conjugate 3, the conjugate using anti-CAPRIN-1 polyclonal antibody #4: SN38 conjugate 4, the conjugate using anti-CAPRIN-1 polyclonal antibody #5: SN38 conjugate 5, and the conjugate using anti-CAPRIN-1 polyclonal antibody #6: SN38 conjugate 6). Furthermore, using the same procedure as above, a solution containing a conjugate with CL2A-SN38 (SN38 control conjugate 1) was obtained using the rabbit control antibody described in Example 1 that does not react with the CAPRIN-1 protein in the same manner.

[0235] Furthermore, using the same method as above, a solution containing a conjugate with CL2A-SN38 (SN38 conjugate 10) was obtained using humanized antibody #4, an anti-CAPRIN-1 monoclonal antibody described in Example 2.

[0236] Furthermore, a conjugate of humanized antibody #1, an anti-CAPRIN-1 monoclonal antibody described in Example 2, and CL2A-SN38 was prepared according to a standard method. Purified humanized antibody #1 was dissolved in PBS(-) at a concentration of 10 mg / mL to prepare a solution. After replacing the buffer with PBS(-), the solution was filtered using a 0.22 μm sterilizing filter membrane to obtain a solution containing humanized antibody #1-CL2A-SN38 of the present invention (SN38 conjugate 7). Similarly, solutions containing conjugates using SN38 were obtained for humanized antibodies #2, #3, and #5 to #74 (SN38 conjugate 8, SN38 conjugate 9, SN38 conjugate 11 to SN38 conjugate 80).

[0237] Furthermore, a solution containing a conjugate with the SN38 linker (SN38 control conjugate 2) was obtained in the same manner as above using a human IgG control antibody that does not react with the CAPRIN-1 protein.

[0238] The number of SN38 molecules bound to each antibody molecule of the above conjugate was measured by mass spectrometry according to a standard method, and the average number was approximately 4 to 5.

[0239] Example 4 Specific Reactivity of Conjugate with CAPRIN-1 Protein and CAPRIN-1-Expressing Cancer Cells The conjugate (anti-CAPRIN-1 antibody-CL2A-SN38 conjugate) prepared in Example 3 was evaluated for its specific reactivity with CAPRIN-1 protein and its reactivity on the cell membrane surface of human cancer cells and mouse cancer cells expressing CAPRIN-1 protein.

[0240] Specific reactivity to the CAPRIN-1 protein was confirmed using ELISA. 100 μL of a 1 μg / mL CAPRIN-1 protein solution was added per well of a 96-well plate and incubated at 4°C for 18 hours. After washing each well three times with PBS-T, 400 μL of 0.5% bovine serum albumin (BSA) solution was added per well and incubated at room temperature for 3 hours. The solution was removed, and the wells were washed three times with 400 μL of PBS-T. Then, 100 μL of each solution containing conjugates 1 to 6 and control conjugate 1 was added per well and incubated at room temperature for 2 hours. After washing each well three times with PBS-T, 100 μL of HRP-labeled anti-rabbit IgG antibody diluted 5000-fold with PBS was added per well and incubated at room temperature for 1 hour. After washing the wells three times with PBS-T, 100 μL of TMB substrate solution was added per well and allowed to stand for 15 to 30 minutes for a color reaction. After color development, 100 μL of 1 N sulfuric acid was added per well to stop the reaction, and absorbance values ​​at 450 nm and 595 nm were measured using an absorbance meter. As a result, SN38 conjugates 1 to 6 had higher absorbance values ​​than the negative control SN38 control conjugate 1, confirming that they react specifically with the CAPRIN-1 protein.

[0241] Similarly, specific reactions to the CAPRIN-1 protein were detected by ELISA using HRP-labeled anti-human antibodies for anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 7 to SN38 conjugates 80) using humanized antibodies #1 to #74 prepared in Example 3. As a result, the absorbance values ​​were higher than those of human IgG (SIGMA) and SN38 control conjugate 2 used as negative controls, confirming that the conjugates react specifically with the CAPRIN-1 protein.

[0242] Next, the reactivity of the CAPRIN-1 protein to the cell membrane surface of cancer cells expressing the protein on the cell surface 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, Becton, Dickinson and Company) to stain dead cells, and then the fluorescence intensity was measured using a FACSFortessa™ (Becton, Dickinson and Company). The results showed that the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (conjugates 1 to 6) had higher fluorescence intensity than the negative control, control conjugate 1, confirming that they strongly react with the cell surface of human breast cancer cell line BT474 and mouse breast cancer cell line 4T1, which express CAPRIN-1 protein on their cell surface.

[0243] Similarly, for anti-CAPRIN-1 antibody-CL2A-SN38 conjugates using humanized antibodies #1 to #74, specific reactivity with CAPRIN-1-expressing cancer cells, human cancer cell BT474 and mouse cancer cell 4T1, was detected using Alexa488-labeled anti-human IgG (H+L) antibody. As a result, it was confirmed that anti-CAPRIN-1 antibody-CL2A-SN38 conjugates using humanized antibodies #1 to #80 (SN38 conjugates 7 to SN38 conjugates 80) had higher fluorescence intensity than the negative control control conjugate 2, i.e., they strongly reacted with the cell surfaces of human breast cancer cell BT474 and mouse breast cancer cell 4T1, which express CAPRIN-1 protein.

[0244] Similarly, the reactivity of anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 7 to SN38 conjugates 80) prepared using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 with various human and mouse cancer cells was confirmed. Human cancer cells in which expression of the CAPRIN-1 gene has been confirmed and expression of the CAPRIN-1 protein on the cell membrane surface of cancer cells has been confirmed, including breast cancer cells (BT-474), colon cancer cells (HT-29, HCT116, and Lovo), lung cancer cells (A549), gastric cancer cells (NCI-N87, NU-GC-3), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (HepG2), ovarian cancer cells (MCAS, SKOV3), renal cancer cells (Caki-2), brain cancer cells (U-87MG), bladder cancer cells (T24, UMUC3), bile duct cancer cells (KKU213), fibrosarcoma cells (HT-1080), and esophageal cancer cells (OE33); When tested on leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB), melanoma cells (Malme-3M), head and neck cancer cells (FaDu), and mouse cancer cells in which expression of CAPRIN-1 protein on the cell membrane surface of cancer cells, namely mouse renal cancer cells (Renca) and mouse breast cancer cells (4T1), the anti-CAPRIN-1 antibody-CL2A-SN38 conjugate prepared in Example 3 had stronger fluorescence intensity than the negative controls SN38 control conjugate 1 and SN38 control conjugate 2, and it was confirmed that it reacts strongly with the cell membrane surface of cancer cells in which CAPRIN-1 protein is expressed.

[0245] Example 5 Internalization of SN38 Conjugates The internalization of anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 1 to SN38 conjugates 80) using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 into cancer cells was confirmed using a confocal laser microscope. HT29 cancer cells, which had been cultured overnight in a culture plate, were incubated with the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates at 100 μg / mL for 1 hour on ice, and then incubated at 37°C and 5% CO for 1 to 6 hours. ２ The cells were reacted under the same conditions. After the reaction, the cells were washed with PBS and fixed at room temperature using PBS containing 4% paraformaldehyde. The fixed cells were then washed with PBS and then subjected to a cell membrane permeabilization treatment using a PBS solution containing 0.3% Triton-X. After permeabilization, the cells were washed with PBS and subjected to a blocking treatment using a PBS solution containing 5% BSA. After blocking, the cells were reacted with an Alexa488-labeled anti-rabbit IgG secondary antibody or anti-human IgG secondary antibody (A11013, Invitrogen) at a concentration of 10 μg / ml for 1 hour at room temperature. After the secondary antibody reaction, the above samples were washed with PBS and then internalization of the conjugate of the present invention into cancer cells was observed using a confocal laser microscope (Nicon A1R inverted microscopy).

[0246] As a result of the observation, the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 1 to SN38 conjugates 80) using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 were clearly present intracellularly, indicating that the conjugates of the present invention were internalized. On the other hand, no internalization into cancer cells was observed for control conjugates 1 and 2 prepared in Example 3.

[0247] (Example 6) Antitumor Activity of SN38 Conjugates The antitumor activity of anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 1 to SN38 conjugates 80) using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 3 against cancer cells was evaluated in vitro. In Example 4, the expression of CAPRIN-1 protein on the cell membrane surface of human cancer cells was confirmed, including breast cancer cells (BT-474), colon cancer cells (HT-29, HCT116), lung cancer cells (NCI-H322, NCI-H2228), gastric cancer cells (NCI-N87, NU-GC-3), uterine cancer cells (Ishikawa, HEC-1-A), and pancreatic cancer cells (Panc10.5). The antitumor activity was evaluated against ovarian cancer cells (MCAS), head and neck cancer cells (FaDu), bladder cancer cells (UMUC3), prostate cancer cells (22Rv1), liver cancer cells (HepG2), kidney cancer cells (Caki-2), brain cancer cells (U-87MG), cholangiocarcinoma cells (KKU213), fibrosarcoma cells (HT-1080), gallbladder cancer cells (TGBC14TKB), and melanoma cells (Malme-3M).

[0248] The cancer cells were cultured by a standard method and plated in a black 96-well plate suitable for cell culture (96 well optical Black Plate Polymer Base Black with Lid Cell Culture Sterile PS 165305, Thermo) at 1 to 4 x 10 cells per well. ４ The cells were seeded at 90 μL / well and incubated in 5% CO ２ The cells were cultured in an incubator at 37°C for 18 to 24 hours. －２ μg / mL to 10 ２ The anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (SN38 conjugates 1 to SN38 conjugates 80) prepared in Example 3 were added at 10 μL per well, and the conjugates were incubated in a 5% CO atmosphere. ２The plates were incubated in an incubator at 37°C. Three to four days after the start of incubation, cell viability was measured using a Cell Titer Glow Luminescent Cell Viability Assay (#7573, Promega). 100 μL of substrate was added per well, and the plates were shaken for 2 minutes using a plate shaker. After allowing to stand for 10 minutes, the luminescence signal intensity was measured using a SpectraMax® iD3 (MOLECULAR DEVICE) to calculate the ATP content of surviving cells.

[0249] As a result, for the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates (conjugates 1 to 80) using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 described in the present invention, the number of surviving cancer cells in a well to which no conjugate was added was 100%, and at concentrations of 1 μg / mL or higher, the number of surviving cancer cells was 70% or less. These results demonstrate that the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates exhibit antitumor activity against cancer cells.

[0250] (Example 7) Antitumor Effect of Conjugates in Cancer-Bearing Mice The antitumor effects of SN38 Conjugate 1 and SN38 Conjugate 7 (both of which have approximately four SN38 molecules bound per antibody molecule), which are the anti-CAPRIN-1 antibody-CL2A-SN38 conjugates prepared in Example 3, were evaluated in vivo in cancer-bearing mice. The antitumor effect of the conjugate of the present invention was examined using SCID mice transplanted with human-derived cancer cells expressing CAPRIN-1 protein. 10 per mouse ７ Human head and neck cancer cells, FaDu, were mixed with Matrigel (SIGMA) and implanted subcutaneously. ３Tumor-bearing mice were prepared by growing the mice to a tumor size of 100 or more. FaDu expresses CAPRIN-1 protein on the cell membrane surface, and as shown in Example 4, SN38 conjugate 7 and SN38 conjugate 10 bind to FaDu. SN38 conjugate 7 and SN38 conjugate 10 were administered to FaDu tumor-bearing mice via the tail vein at 20 mg / kg once a week for a total of eight doses.

[0251] Furthermore, a conjugate of CL2A-SN38 with trastuzumab (Chugai Pharmaceutical Co., Ltd.), an anti-HER2 antibody, was prepared according to the method described in Example 3 so that the number of drugs bound per antibody molecule was similar to that of SN38 Conjugate 1 and SN38 Conjugate 7, and the same amount was administered to the above-mentioned tumor-bearing mice as a comparative control. In FaDu, HER2 protein, the target antigen of trastuzumab, is expressed on the cell membrane surface, and the above-mentioned conjugate of trastuzumab with a topoisomerase I inhibitor specifically binds to it.

[0252] After administration, the size of the tumor in the tumor-bearing mice was measured over time using a vernier caliper, and the tumor volume was calculated according to the standard method using the formula: (length of the longest axis of the tumor) x (length of the shortest axis of the tumor). ２ As a result of the evaluation, on day 7 after the start of administration, the tumor volume of the cancer-bearing mice administered the above-mentioned conjugate was less than 60%, assuming that the tumor volume of the cancer-bearing mice administered the anti-CAPRIN-1 antibody not conjugated with SN38 (control) was 100%.

[0253] On the other hand, the tumor volume of the mice administered with the trastuzumab-CL2A-SN38 conjugate was approximately 75% of the tumor volume of the tumor-bearing mice administered with trastuzumab not conjugated with SN38, which was taken as 100%.

[0254] The results of this evaluation revealed that the anti-CAPRIN-1 antibody-CL2A-SN38 conjugate had a significantly enhanced anti-tumor effect compared to an anti-CAPRIN-1 antibody not conjugated with SN38 (control), whereas the anti-tumor effect of the trastuzumab-CL2A-SN38 conjugate was only slightly enhanced compared to trastuzumab not conjugated with SN38.

[0255] (Example 8) PEG ９ Synthesis of EVA-PAB-Exatecan: As a topoisomerase I inhibitor conjugated to an antibody, exatecan is a compound having the following chemical structure, (S)-1-((2,5-dioxopyrrolidin-1-yl)oxy)-33-(((S)-1-(((S)-1-((4-((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H, 12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-1,31-dioxo-4,7,10,13,16,19,22,25,28-nonaoxa-32-azahexatriacontan-36-oic acid (hereinafter referred to as "PEG ９ -EVA-PAB-Exatecan) was synthesized.

[0256]

[0257] Specifically, 4-((S)-2-((S)-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)benzyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3'4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (CAS: 2943010-48-6, 9.2 mg, 0.0094 mmol) was dissolved in dimethylformamide (270 μL), and then diethylamine (30 μL) was added and stirred at room temperature for 2 hours. The reaction solution was evaporated under reduced pressure, and then 5-tert-butyl N-(tert-butoxycarbonyl)-L-glutamate (4.3 mg, 0.0141 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (5.2 mg, 0.0094 mmol), and 5-tert-butyl N-(tert-butoxycarbonyl)-L-glutamate (4.3 mg, 0.0141 mmol) were added. A solution of 4,7,10,13,16,19,22,25,28-nonaoxahentriacontanedioate (N-succinimidyl) (13.3 mg, 0.0136 mmol) and diisopropylethylamine (4.7 μL, 0.0273 mmol) in dimethylformamide (200 μL) was added and stirred at room temperature for 1 hour. The reaction solution was then evaporated under reduced pressure and purified by silica gel column chromatography. The resulting compound was dissolved in dichloromethane (150 μL), and trifluoroacetic acid (150 μL) was added under ice cooling. The mixture was then stirred under ice cooling for 4.5 hours and the reaction solution was then evaporated under reduced pressure. Half of the resulting residue (0.0047 mmol) was diluted with 4,7,10,13,16,19,22,25,28-nonaoxahentriacontanedioate di(N-succinimidyl) (13.3 After adding PEG-100 (mg, 0.0188 mmol) and dimethylformamide (200 μL), triethylamine (3.9 μL, 0.0282 mmol) was added under ice cooling. The mixture was stirred at room temperature for 1 hour, and the reaction mixture was evaporated under reduced pressure. The PEG-100 was purified by high performance liquid chromatography (HPLC) (Nexera Prep, Shimadzu Corporation). ９ -EVA-PAB-Exatecan (1.5 mg) was obtained.

[0258] (Example 9) Preparation of conjugates of anti-CAPRIN-1 antibodies and topoisomerase I inhibitors (exatecan) Conjugates of anti-CAPRIN-1 antibodies and topoisomerase I inhibitors were prepared using anti-CAPRIN-1 polyclonal antibodies #1 to #6 described in Example 1 and PEG-1 antibodies containing exatecan as the antitumor active ingredient described in Example 8. ９ -EVA-PAB-Exatecan conjugate (Exatecan conjugate) was prepared according to a standard method.

[0259] The purified anti-CAPRIN-1 polyclonal antibody #1 was dissolved in PBS(-) at a concentration of 2.5 mg / mL. The PEG prepared in Example 8 was added to the solution containing the polyclonal antibody #1. ９ 78 μL of a DMSO solution containing 10 mM EVA-PAB-Exatecan was added to the mixture, and the mixture was left to stand at room temperature for 1 hour. The reaction solution was desalted twice using a Zeba spin column (40K MWCO) substituted with PBS, and then filtered through a 0.22 μm filter to obtain the anti-CAPRIN-1 polyclonal antibody #1-PEG of the present invention. ９ A solution containing EVA-PAB-Exatecan (Exatecan Conjugate 1) was obtained.

[0260] Similarly, anti-CAPRIN-1 polyclonal antibodies #2 to #6 were also PEG- ９ A solution containing an exatecan conjugate using EVA-PAB-Exatecan was obtained (the conjugate using anti-CAPRIN-1 polyclonal antibody #2: Exatecan Conjugate 2, the conjugate using anti-CAPRIN-1 polyclonal antibody #3: Exatecan Conjugate 3, the conjugate using anti-CAPRIN-1 polyclonal antibody #4: Exatecan Conjugate 4, the conjugate using anti-CAPRIN-1 polyclonal antibody #5: Exatecan Conjugate 5, and the conjugate using anti-CAPRIN-1 polyclonal antibody #6: Exatecan Conjugate 6). Furthermore, using the same procedure as above, the rabbit control antibody described in Example 1, which does not react with the CAPRIN-1 protein, was also prepared by the same method using PEG.９ A solution containing a conjugate of EVA-PAB-Exatecan (Exatecan Control Conjugate 1) was obtained.

[0261] In addition, using the same method as above, humanized antibody #4, which is the anti-CAPRIN-1 monoclonal antibody described in Example 2, was used to prepare a PEG- ９ A solution containing a conjugate of EVA-PAB-Exatecan (Exatecan Conjugate 10) was obtained.

[0262] In addition, humanized antibody #1, which is an anti-CAPRIN-1 monoclonal antibody described in Example 2, and PEG ９ A conjugate of the humanized antibody #1 of the present invention with EVA-PAB-Exatecan was prepared according to a standard method. The purified humanized antibody #1 was dissolved in PBS(-) at a concentration of 10 mg / mL. After replacing the buffer with PBS(-), the solution was filtered using a 0.22 μm sterilizing filter membrane to obtain the humanized antibody #1-PEG conjugate of the present invention. ９ A solution containing EVA-PAB-Exatecan (Exatecan Conjugate 7) was obtained. Similarly, solutions containing conjugates using exatecan were obtained for humanized antibodies #2, #3, and #5 to #74 (Exatecan Conjugate 8, Exatecan Conjugate 9, Exatecan Conjugate 11 to Exatecan Conjugate 80).

[0263] Furthermore, a solution containing a conjugate with an exatecan linker (exatecan control conjugate 2) was obtained in the same manner as above using a human IgG control antibody that does not react with the CAPRIN-1 protein.

[0264] The number of exatecan molecules bound to each antibody molecule of the conjugate was measured using an ultraviolet spectrophotometer (NanoDrop One, Thermo Scientific) according to a standard method, and the average number was found to be approximately 4-5.

[0265] Example 10: Specific reactivity of exatecan conjugate to CAPRIN-1 protein and CAPRIN-1-expressing cancer cells. The exatecan conjugate (anti-CAPRIN-1 antibody-PEG) prepared in Example 8 was ９ The specific reactivity of the CAPRIN-1 protein (E2-EVA-PAB-Exatecan conjugate) and its reactivity on the cell membrane surface of human cancer cells and mouse cancer cells expressing the CAPRIN-1 protein were evaluated.

[0266] Specific reactivity to CAPRIN-1 protein was confirmed using ELISA. 100 μL of 1 μg / mL CAPRIN-1 protein solution was added per well of a 96-well plate and incubated at 4°C for 18 hours. After washing each well three times with PBS-T, 400 μL of 0.5% bovine serum albumin (BSA) solution was added per well and incubated at room temperature for 3 hours. The solution was removed, and the wells were washed three times with 400 μL of PBS-T. Then, 100 μL of solutions containing exatecan conjugates 1 to 6 and exatecan control conjugate 1 were added per well and incubated at room temperature for 2 hours. After washing each well three times with PBS-T, 100 μL of HRP-labeled anti-rabbit IgG antibody diluted 5000-fold with PBS was added per well and incubated at room temperature for 1 hour. After washing the wells three times with PBS-T, 100 μL of TMB substrate solution was added per well and the plate was allowed to stand for 15 to 30 minutes to allow the color reaction to occur. After color development, 100 μL of 1 N sulfuric acid was added per well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorption spectrometer. As a result, exatecan conjugates 1 to 6 had higher absorbance values ​​than the negative control, exatecan control conjugate 1, confirming that they react specifically with the CAPRIN-1 protein.

[0267] Similarly, anti-CAPRIN-1 antibody-PEG was prepared using the humanized antibodies #1 to #74 prepared in Example 3. ９For the EVA-PAB-Exatecan conjugates (Exatecan Conjugates 7 to 80), a specific reaction with the CAPRIN-1 protein was detected by ELISA using an HRP-labeled anti-human antibody. As a result, the absorbance values ​​were higher than those of human IgG (SIGMA) used as a negative control and Exatecan Control Conjugate 2, confirming that the conjugates react specifically with the CAPRIN-1 protein.

[0268] Next, the reactivity of the CAPRIN-1 protein to the cell membrane surface of cancer cells expressing the protein on the cell surface was confirmed by flow cytometry. ５ Human 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 exatecan conjugates 1 to 6 and exatecan 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, Becton, Dickinson and Company) to stain dead cells, and then the fluorescence intensity was measured using a FACSFortessa™ (Becton, Dickinson and Company). ９ The EVA-PAB-Exatecan conjugates (Exatecan conjugates 1 to 6) had higher fluorescence intensities than the negative control Exatecan control conjugate 1, and it was confirmed that they reacted strongly with the cell surfaces of human breast cancer cell BT474 and mouse breast cancer cell 4T1, on whose cell surfaces CAPRIN-1 protein is expressed.

[0269] Similarly, anti-CAPRIN-1 antibody-PEG using humanized antibodies #1 to #74 ９The specific reactivity of the EVA-PAB-Exatecan conjugate with CAPRIN-1-expressing cancer cells, human cancer cell BT474 and mouse cancer cell 4T1, was detected using Alexa488-labeled anti-human IgG (H+L) antibody. As a result, the anti-CAPRIN-1 antibody-PEG conjugate using humanized antibodies #1 to #80 ９ The EVA-PAB-Exatecan conjugates (Exatecan conjugates 7 to 80) had higher fluorescence intensities than the negative control, Exatecan control conjugate 2, and were therefore confirmed to react strongly with the cell surfaces of human breast cancer cells BT474 and mouse breast cancer cells 4T1, which express CAPRIN-1 protein.

[0270] Similarly, anti-CAPRIN-1 antibody-PEG using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 8 ９ The reactivity of EVA-PAB-Exatecan conjugates (Exatecan Conjugate 7 to Exatecan Conjugate 80) with various human and mouse cancer cells was confirmed. These human cancer cells are confirmed to express the CAPRIN-1 gene and to have CAPRIN-1 protein on the cell membrane surface of cancer cells, including breast cancer cells (BT-474), colon cancer cells (HT-29, HCT116, and Lovo), lung cancer cells (A549), gastric cancer cells (NCI-N87, NU-GC-3), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (HepG2), ovarian cancer cells (MCAS, SKOV3), renal cancer cells (Caki-2), and brain cancer cells (U-87). MG), bladder cancer cells (T24, UMUC3), bile duct cancer cells (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), and mouse renal cancer cells (Renca) and mouse breast cancer cells (4T1), which are mouse cancer cells in which expression of CAPRIN-1 protein on the cell membrane surface of cancer cells has been confirmed.９ The EVA-PAB-Exatecan conjugates both had stronger fluorescence intensities than the negative controls, Exatecan Control Conjugate 1 and Exatecan Control Conjugate 2, and were confirmed to react strongly with the cell membrane surface of cancer cells expressing CAPRIN-1 protein.

[0271] (Example 11) Internalization of exatecan conjugates Anti-CAPRIN-1 antibody-PEG using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 8 ９ The internalization of the EVA-PAB-Exatecan conjugates (Exatecan Conjugate 1 to Exatecan Conjugate 80) into cancer cells was confirmed using a confocal laser microscope. The anti-CAPRIN-1 antibody-CL2A-SN38 conjugate was reacted at 100 μg / mL with HT29 cancer cells, which had been cultured overnight in a culture plate, on ice for 1 hour, and then incubated at 37°C and 5% CO for 1 to 6 hours. ２ The cells were reacted under the same conditions. After the reaction, the cells were washed with PBS and fixed at room temperature using PBS containing 4% paraformaldehyde. The fixed cells were then washed with PBS and then subjected to a cell membrane permeabilization treatment using a PBS solution containing 0.3% Triton-X. After permeabilization, the cells were washed with PBS and subjected to a blocking treatment using a PBS solution containing 5% BSA. After blocking, the cells were reacted with an Alexa488-labeled anti-rabbit IgG secondary antibody or anti-human IgG secondary antibody (A11013, Invitrogen) at a concentration of 10 μg / ml for 1 hour at room temperature. After the secondary antibody reaction, the above samples were washed with PBS and then internalization of the conjugate of the present invention into cancer cells was observed using a confocal laser microscope (Nicon A1R inverted microscopy).

[0272] As a result of the observation, the anti-CAPRIN-1 antibody-PEG mixture using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 8 was ９The EVA-PAB-Exatecan conjugates (Exatecan conjugates 1 to 80) were clearly present within the cells, indicating that the exatecan conjugates of the present invention were internalized. On the other hand, no internalization into cancer cells was observed for the exatecan control conjugates 1 and 2 prepared in Example 8.

[0273] (Example 12) Antitumor activity of exatecan conjugates Anti-CAPRIN-1 antibody-PEG conjugates using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 8 ９ The antitumor activity of EVA-PAB-Exatecan conjugates (Exatecan Conjugate 1 to Exatecan Conjugate 80) against cancer cells was evaluated in vitro. In Example 4, the expression of CAPRIN-1 protein on the cell membrane surface of human cancer cells was confirmed, including breast cancer cells (BT-474), colon cancer cells (HT-29, HCT116), lung cancer cells (NCI-H322, NCI-H2228), gastric cancer cells (NCI-N87), uterine cancer cells (Ishikawa, HEC-1-A), pancreatic cancer cells (Panc10.5), and ovarian cancer cells (MCA). S), head and neck cancer cells (FaDu), gastric cancer (NU-GC-3), bladder cancer (UMUC3), prostate cancer cells (22Rv1), liver cancer cells (HepG2), renal cancer cells (Caki-2), brain tumor cells (U-87MG), cholangiocarcinoma cells (KKU213), fibrosarcoma cells (HT-1080), gallbladder cancer cells (TGBC14TKB), and melanoma cells (Malme-3M) were evaluated for antitumor activity.

[0274] The cancer cells were cultured by a standard method and plated in a black 96-well plate suitable for cell culture (96 well optical Black Plate Polymer Base Black with Lid Cell Culture Sterile PS 165305, Thermo) at 1 to 4 x 10 cells per well. ４ The cells were seeded at 90 μL / well and incubated in 5% CO ２ The cells were cultured in an incubator at 37°C for 18 to 24 hours. －３ μg / mL to 10 ２The anti-CAPRIN-1 antibody-PEG mixture was prepared using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 prepared in Example 8 in a range of μg / mL. ９ -EVA-PAB-Exatecan conjugates (Exatecan conjugate 1 to Exatecan conjugate 80) were added at 10 μL per well, and the plates were incubated in 5% CO ２ The plates were incubated in an incubator at 37°C. Three to four days after the start of incubation, cell viability was measured using a Cell Titer Glow Luminescent Cell Viability Assay (#7573, Promega). 100 μL of substrate was added per well, and the plates were shaken for 2 minutes using a plate shaker. After allowing to stand for 10 minutes, the luminescence signal intensity was measured using a SpectraMax® iD3 (MOLECULAR DEVICE) to calculate the ATP content of surviving cells.

[0275] As a result, anti-CAPRIN-1 antibody-PEG synthesis using the anti-CAPRIN-1 polyclonal antibody and humanized antibodies #1 to #74 described in the present invention was achieved. ９ With the anti-CAPRIN-1 antibody-PEG conjugates (Exatecan Conjugate 1 to Exatecan Conjugate 80), the survival rate of cancer cells in wells without conjugates was 70% or less at concentrations of 1 μg / mL or higher, assuming that the survival rate of cancer cells in wells without conjugates was 100%. ９ It was revealed that the EVA-PAB-Exatecan conjugate exhibits antitumor activity against cancer cells.

[0276] (Example 13) Antitumor effect of exatecan conjugate in tumor-bearing mice Anti-CAPRIN-1 antibody-PEG prepared in Example 8 ９The antitumor effects of the EVA-PAB-Exatecan conjugates Exatecan Conjugate 1 and Exatecan Conjugate 10 (both of which have approximately four molecules of Exatecan bound to one antibody molecule) in vivo in tumor-bearing mice were evaluated. The antitumor effects of the conjugates of the present invention were examined using SCID mice transplanted with human-derived cancer cells expressing CAPRIN-1 protein. ７ Human colon cancer cells HT-29 were mixed with Matrigel (Corning) and subcutaneously implanted, and the tumors grew to 120 mm ３ Tumor-bearing mice were prepared by growing the tumor to a tumor size of 100 or larger. HT-29 expresses CAPRIN-1 protein on the cell membrane surface, and as shown in Example 4, exatecan conjugate 1 and exatecan conjugate 10 bind to HT-29. Exatecan conjugate 1 and exatecan conjugate 10 were administered to HT-29 tumor-bearing mice via the tail vein at 10 mg / kg twice a week, a total of four times.

[0277] In addition, according to the method described in Example 8, trastuzumab (Selleck), an anti-HER2 antibody, and PEG ９ A conjugate of EVA-PAB-Exatecan was prepared so that the number of drugs bound per antibody molecule was similar to that of Exatecan Conjugate 1 and Exatecan Conjugate 10 (4 to 5), and the same amount was administered to the above-mentioned tumor-bearing mice as a control. HT-29 expresses HER2 protein, the target antigen of trastuzumab, on the cell membrane surface, to which the above-mentioned conjugate of trastuzumab and a topoisomerase I inhibitor specifically binds.

[0278] After administration, the size of the tumor in the tumor-bearing mice was measured over time using a vernier caliper, and the tumor volume was calculated according to the standard method using the formula: (length of the longest axis of the tumor) x (length of the shortest axis of the tumor). ２ The evaluation results showed that on day 10 after the start of administration, the tumor volume of the tumor-bearing mice administered the above-mentioned conjugate was less than 55%, assuming that the tumor volume of the tumor-bearing mice administered an anti-CAPRIN-1 antibody not conjugated with exatecan (control) was 100%.

[0279] On the other hand, trastuzumab-PEG ９ The tumor volume of the mice administered with the EVA-PAB-Exatecan conjugate was approximately 65% ​​of the tumor volume of the tumor-bearing mice administered with trastuzumab not conjugated with Exatecan, which was taken as 100%.

[0280] As a result of this evaluation, anti-CAPRIN-1 antibody-PEG ９ The EVA-PAB-Exatecan conjugate showed enhanced antitumor effects compared to the non-exatecan-conjugated anti-CAPRIN-1 antibody (control), whereas the trastuzumab-PEG conjugate showed no significant effect. ９ It was revealed that the antitumor effect of the EVA-PAB-Exatecan conjugate was weaker than that of trastuzumab not conjugated with Exatecan.

Claims

1. A conjugate comprising an antibody or a fragment thereof immunologically reactive with a CAPRIN-1 protein having an amino acid sequence represented by any of the even-numbered SEQ ID NOS: 2 to 30, or an amino acid sequence having 80% or more sequence identity with said amino acid sequence, and a topoisomerase I inhibitor bound thereto.

2. The conjugate according to claim 1, wherein the antibody or fragment thereof is immunologically reactive with a partial polypeptide of a CAPRIN-1 protein having 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 80% or more sequence identity with said amino acid sequence.

3. The conjugate of claim 1, wherein the antibody is a monoclonal or polyclonal antibody.

4. The conjugate according to claim 1, wherein the antibody or fragment thereof is any one of the following (A) to (M): (A) an antibody or fragment comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 36, 37, and 38 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 40, 41, and 42 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein; or (B) an antibody or fragment comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 44, 45, and 46 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 48, 49, and 50 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (C) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 52, 53, and 54 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 56, 57, and 58 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with the CAPRIN-1 protein. (D) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 60, 61, and 62 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 64, 65, and 66 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with the CAPRIN-1 protein. (E) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (F) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein.(G) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (H) An antibody or fragment thereof comprising 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 having immunological reactivity with CAPRIN-1 protein. (I) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 146, 147, and 148 (CDR1, CDR2, and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 149, 150, and 151 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (J) An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 272, 273, and 274 (CDR1, CDR2, and CDR3, respectively), and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 275, 276, and 277 (CDR1, CDR2, and CDR3, respectively), and having immunological reactivity with CAPRIN-1 protein. (K) 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: 290, 291 and 292 and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 293, 294 and 295, and having immunological reactivity with the CAPRIN-1 protein. (L) 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: 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, and having immunological reactivity with the CAPRIN-1 protein.(M) An antibody or fragment thereof comprising a heavy chain variable region containing the complementarity determining regions (CDR1, CDR2 and CDR3) of SEQ ID NOs: 134, 135 and 136, and a light chain variable region containing the complementarity determining regions (CDR1, CDR2 and CDR3) of SEQ ID NOs: 137, 138 and 139, and having immunological reactivity with a CAPRIN-1 protein.

5. The conjugate according to claim 1, wherein the antibody or fragment thereof is any one of the following (a) to (al): (a) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 43; (b) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 47 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 51; (c) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; (d) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 67; (e) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 69; or (f) an antibody or fragment thereof whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

71. (g) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 73; (h) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 75; (i) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 77; (j) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 79; (k) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 81; (l) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

83. (m) an antibody or 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 fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 87; (o) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 89; (p) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 91; (q) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 92 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 93; (r) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 94 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 95; (s) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 96 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

97. (t) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 99; (u) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 101; (v) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 103; (w) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 105; (x) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 107; (y) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

109. (z) an antibody or 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 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 fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 115; (ac) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 117; (ad) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 119; (ae) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 121; (af) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 123; (ag) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

125. (ah) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 127; (ai) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 129; (aj) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 131; (ak) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO: 133; (al) an antibody or fragment thereof, whose heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and whose light chain variable region comprises the amino acid sequence of SEQ ID NO:

304.

6. The conjugate of claim 1, 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 and the topoisomerase I inhibitor are linked via a linker.

8. The conjugate of claim 1, wherein the topoisomerase I inhibitor is exatecan, SN38, or a derivative thereof.

9. A pharmaceutical composition for treating and / or preventing cancer, comprising as an active ingredient the conjugate according to any one of claims 1 to 8.

10. The pharmaceutical composition according to claim 9, wherein the cancer is a cancer that expresses CAPRIN-1 protein on the cell membrane surface.

11. The pharmaceutical composition of claim 10, wherein the cancer is breast cancer, kidney cancer, pancreatic cancer, colon 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 cancer.

12. A method for treating and / or preventing cancer, comprising administering to a subject the conjugate according to any one of claims 1 to 8 or the pharmaceutical composition according to any one of claims 9 to 11.

Images