Specification
Title: Pharmaceutical composition for treating and / or preventing cancer
Technical field
[0001]
　The present invention relates to a conjugate of an antibody against CAPRIN-1 protein and an immunostimulatory agent, and a pharmaceutical use as a therapeutic and / or prophylactic agent for cancer using the same.
Background art
[0002]
　Because of their cancer specificity, various antibody drugs that target specific antigen proteins on cancer cells are applied to cancer treatment as cancer treatment drugs with few side effects. For example, the cell membrane surface of many solid cancers expresses Cytosmic-activation and proliferation-associated protein 1 (CAPRIN-1), and an antibody against this CAPRIN-1 protein is used as a pharmaceutical for treating and / or preventing cancer. It is known to be promising as (Patent Document 1).
[0003]
　In recent years, studies are underway to enhance the efficacy of these antibody drugs, and in particular, development of antibody-drug conjugates (ADC) in which a drug and an antibody have been directly conjugated to cells with strong killing ability Advanced (Non-Patent Documents 1 and 2). As an example of ADC, Trastuzumab (Trastuzumab emtansine) in which a drug having an ability to kill cells is linked to trastuzumab, which is an existing antibody drug, (Trastuzumab emtansine), or an anti-CD30 monoclonal antibody as monomethyl auristatin E (MMAE) (Brentuximab vedotin) is used for the treatment of some cancers, and the survival rate is shown to be extended compared to conventional treatments, and its usefulness for existing cancer treatments is It is shown (nonpatent literature 3 and 4).
[0004]
　In another example, studies have also been conducted to conjugate an immunostimulatory agent to an antibody drug against cancer to enhance its efficacy. For example, a conjugate obtained by binding Resiquimod, one of the immunostimulating agents, to the existing antibody drugs trastuzumab and cetuximab, or a conjugate of rituximab, which is an antibody drug targeting CD20 as a target antigen, with various immunostimulating agents Thus, the efficacy enhancing effect of antibodies has been recognized in animal models (Patent Documents 2 and 3).
[0005]
　As described above, by conjugating various factors with various antibodies, attempts have been made to enhance the efficacy of antibody drugs against cancer, but a potent antitumor effect is obtained that causes a complete regression of various cancers. It has not been done. In addition, the effect of preventing cancer recurrence and metastasis has not been shown.
Prior art documents
Patent document
[0006]
Patent Document 1: WO2010 / 016526
Patent Document 2: WO2014 / 012479
Patent Document 3: US Patent No. 8,951,528
Non-patent literature
[0007]
Non-Patent Document 1: Lancet Oncol 2016; 17: e256-62
Non-Patent Document 2: Pharm Res. 2015 Nov; 32 (11): 3526-40
Non-Patent Document 3: New England Journal of Medicine 467; 19 (8), 2012
Non-Patent Document 4: MAbs 2012; 4 (4): 458-65
Summary of the invention
Problems that the Invention is to Solve
[0008]
　An object of the present invention is to provide a means for enhancing the antitumor effect of an antibody against a CAPRIN-1 protein expressed on the cell membrane surface of cancer cells or a fragment thereof.
Means to solve the problem
[0009]
　The inventors of the present invention have conducted intensive studies to show that an antibody against a CAPRIN-1 protein or a conjugate thereof with an immunostimulatory factor exerts an extremely strong antitumor effect compared to an antibody against a CAPRIN-1 protein or its fragment alone Furthermore, the enhancing effect of the antitumor effect when an immunostimulatory factor is conjugated to an antibody against CAPRIN-1 protein or a fragment thereof is the antitumor effect when an immunostimulatory factor is conjugated to an existing cancer antibody drug The present invention has been found to be significantly superior to the enhancing effect of
[0010]
　Specifically, the present invention has the following features (1) to (14).
(1) Immunologically with a CAPRIN-1 protein having an amino acid sequence represented by any one of SEQ ID NOs: 2 to 30, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence A conjugate comprising an antibody or fragment thereof having reactivity and an immunostimulatory factor bound thereto.
(2) The antibody or fragment thereof has an amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, 309 or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. The conjugate according to (1), which has immunological reactivity with a partial polypeptide of CAPRIN-1 protein.
(3) The conjugate according to (1) or (2), wherein the antibody is a monoclonal antibody or a polyclonal antibody.
(4) The conjugate
(A) of SEQ ID NO: 36, 37 and 38 according to any of (1) to (3), wherein said antibody or fragment thereof is any of the following (A) to (M) : A heavy chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions of SEQ ID NO: 40, 41 and 42 (CDR1, CDR2 and CD3 respectively), and , Antibodies or fragments having immunological reactivity with CAPRIN-1 protein
(B) A heavy chain variable region containing the complementarity determining regions of SEQ ID NO: 44, 45 and 46 (CDR1, CDR2 and CDR3 respectively) and a complementarity determining region of SEQ ID NO: 48, 49 and 50 (CDR1, CDR2 and CDR3 respectively) And an antibody or fragment having immunological reactivity with a CAPRIN-1 protein
(C) complementarity-determining regions of SEQ ID NO: 52, 53 and 54 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 56, 57 and 58, respectively), and having immunological reactivity with CAPRIN-1 protein antibody or fragment having
complementarity determining regions (each of (D) SEQ ID NO: 60, 61 and 62 CDRl, C Immunology with CAPRIN-1 protein, comprising a heavy chain variable region comprising R2 and CDR3) and a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 64, 65 and 66 (CDR1, CDR2 and CDR3 respectively)
Heavy chain variable region containing complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NOS: 170, 171 and 172, respectively) having reactive reactivity with complementarity determining regions of SEQ ID NOS: 173, 174 and 175 An antibody or fragment thereof comprising a light chain variable region comprising (CDR1, CDR2 and CDR3 respectively) and having immunological reactivity with CAPRIN-1 protein
(F) A heavy chain variable region comprising the complementarity determining regions of SEQ ID NO: 176, 177 and 178 (CDR1, CDR2 and CDR3 respectively) and a complementarity determining region of SEQ ID NO: 179, 180 and 181 (CDR1, CDR2 and CDR3 respectively) Antibody or an antibody having an immunological reactivity with the CAPRIN-1 protein, or
the complementarity-determining region of SEQ ID NO: 182, 183 and 184 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 185, 186 and 187, respectively) and immunologically reactive with the CAPRIN-1 protein antibody or fragment thereof having an
(H) SEQ ID NO: 188, 189 and 190 Comprising a heavy chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions of SEQ ID NO: 191, 192 and 193 (CDR1, CDR2 and CDR3 respectively), and
A heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 146, 147 and 148, respectively) of the antibody or fragment thereof (I) having immunological reactivity with CAPRIN-1 protein and SEQ ID NO: 149 , And a light chain variable region comprising the 150 and 151 complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and an antibody or fragment thereof having immunological reactivity with a CAPRIN-1 protein
(J) The heavy chain variable region containing the complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of SEQ ID NO: 272, 273 and 274 and the complementarity determining regions of SEQ ID NO: 275, 276 and 277 (CDR1, CDR2 and CDR3 respectively) Antibody or an antibody having an immunological reactivity with the CAPRIN-1 protein, or a fragment thereof
(K), the complementarity-determining regions of SEQ ID NO: 290, 291 and 292 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 293, 294 and 295 (CDR1, CDR2 and CDR3 respectively) and immunologically reactive with the CAPRIN-1 protein antibody or fragment thereof having an
(L) SEQ ID NO: 301, 302 and 303 A heavy chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of SEQ ID NOs: 305, 306 and 307, and
A heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 134, 135 and 136, respectively) of the antibody or fragment thereof (M) having immunological reactivity with CAPRIN-1 protein and SEQ ID NO: 137 , And a light chain variable region containing complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of 138 and 139, and an antibody having immunological reactivity with CAPRIN-1 protein or a fragment thereof
(5) Or a fragment thereof is any of the following (a) to (ak): 1) conjugate according to any one of the - (4).
(A) An antibody or fragment thereof wherein the
heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43 (b) the heavy chain variable region comprises SEQ ID NO: 47 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 51
(c) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 59 or a fragment thereof
(d) an antibody or 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 fragments thereof
comprise (e) the amino acid sequence of the heavy chain variable region SEQ ID NO: 68 and an antibody or its light chain variable region comprising the amino acid sequence of SEQ ID NO: 69 Fragment
(f) the heavy chain variable region comprises 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 antibody or fragment thereof
(g) the heavy chain variable region SEQ ID NO: 72 Wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73
(h) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74, and the light chain variable region An antibody or fragment thereof
(i) 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 Or a fragment thereof
(J) 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 (k) the heavy chain variable region comprises SEQ ID NO: 80 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81
(1) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82, and the light chain variable region comprises An antibody or fragment thereof
(m) comprising the amino acid sequence of SEQ ID NO: 83, or an antibody or 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 fragments thereof
comprise (n) the amino acid sequence of the heavy chain variable region SEQ ID NO: 86 and an antibody or its light chain variable region comprising the amino acid sequence of SEQ ID NO: 87 Fragment
(o) a heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 89 antibody or fragment thereof
(p) a heavy chain variable region SEQ ID NO: 90 Wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 91
(q) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 92, and the light chain variable region An antibody or fragment thereof
(r) comprising the amino acid sequence of SEQ ID NO: 93, 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 Or a fragment thereof
(S) 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 (t) the heavy chain variable region comprises SEQ ID NO: 98 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 99
(u) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100, and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 101 or a fragment thereof
(v) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 103 fragments thereof
comprise (w) an amino acid sequence of the heavy chain variable region SEQ ID NO: 104 and light chain variable region I comprising the amino acid sequence of SEQ ID NO: 105 Antibody or fragment thereof
(x) a heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106, and antibodies or fragments thereof the light chain variable region comprising the amino acid sequence of SEQ ID NO: 107
(y) heavy chain variable region An antibody comprising the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 109, or a fragment thereof
(z) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and The antibody or fragment thereof
(aa) wherein the chain variable region comprises the amino acid sequence of SEQ ID NO: 111 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 Or a fragment thereof
(Ab) An antibody or fragment thereof in which 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 An antibody or fragment thereof
(ad) comprising the amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117 , and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 119 or a fragment
(ae) 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 fragments thereof
comprise (af) amino acid sequence of the heavy chain variable region SEQ ID NO: 122, and Ami light chain variable region SEQ ID NO: 123 Antibody or fragment thereof comprising an acid sequences
(ag) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124, and antibodies or fragments thereof the light chain variable region comprising the amino acid sequence of SEQ ID NO: 125
(ah An antibody whose 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 or a fragment
(ai) heavy chain variable region thereof 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 or a fragment thereof
(ai) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130, and the light chain variable region comprises the SEQ ID NO: An antibody or fragment thereof comprising the amino acid sequence of 131
(Aj) 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 An antibody or fragment thereof comprising an amino acid sequence and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 304
(6) the antibody is a human antibody, a humanized antibody, a chimeric antibody, a single chain antibody 1) The conjugate in any one of (5).
(7) The immunostimulatory agent is a ligand or agonist that binds to Toll-like receptor (TLR), NOD-like receptor (NLR), RIG-like receptor, C-type lectin receptor (CLR), (1) ) The conjugate according to any one of (6).
(8) The conjugate according to (7), wherein the Toll-like receptor (TLR) is TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9 or TLR10.
(9) The conjugate according to (7) or (8), wherein the ligand or agonist that binds to the Toll-like receptor (TLR) or the derivative thereof is any of the following (i) to (vii):
(I) a ligand or an agonist which binds to TLR2 selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide and zymosan
(ii) selected from the group consisting of Poly (I: C) and poly (A: U) Ligand or agonist that binds to TLR3
(iii) ligand or agonist that binds to TLR4 selected from the group consisting of lipopolysaccharide (LPS), HSP60, RS09 and MPLA
(Iv) a ligand or an agonist which binds to TLR5 selected from the group consisting of flagellin and FLA
(v) a ligand or an agonist which binds to TRL7 or 8 selected from the group consisting of imidazoquinolines compounds and single stranded RNA
(vi) A ligand or agonist that binds to TRL9 selected from the group consisting of bacterial DNA, unmethylated CpG DNA, hemozorin, ODN 1585, ODN 1668, ODN 1668 and ODN 1826
(vii) binds to TRL 10 selected from the group consisting of profilin and uropathogenic bacteria Ligand or agonist
(10) The conjugate according to any one of (1) to (9), wherein the antibody or fragment thereof and the immunostimulatory agent are linked via a linker T.
(11) A pharmaceutical composition for treating and / or preventing cancer, which comprises the conjugate according to any one of (1) to (10) as an active ingredient.
(12) The pharmaceutical composition according to (11), wherein the cancer is a cancer that expresses CAPRIN-1 protein on the cell membrane surface.
(13) The cancer is a breast cancer, a renal cancer, a pancreatic cancer, a colon cancer, a colon cancer, a lung cancer, a brain tumor, a gastric cancer, a uterine cancer, an ovarian cancer, a prostate cancer, a bladder cancer, an esophageal cancer, a leukemia, a lymphoma, a liver cancer, a gallbladder cancer, a sarcoma (11) or a cancer selected from the group consisting of mast cell tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer and head and neck cancer, (11) or The pharmaceutical composition as described in (12).
(14) A method for treating and / or treating cancer, which comprises administering to a subject the conjugate according to any one of (1) to (10) and the pharmaceutical composition according to any of (11) to (13). How to prevent.
Effect of the invention
[0011]
　The conjugate according to the present invention not only exerts a very strong anti-tumor effect as compared to the antibody against CAPRIN-1 protein alone, but is superior to the conjugate of cancer antibody drug and immunostimulatory factor known so far. Have an antitumor effect. Moreover, the enhancement effect of the anti-tumor effect by the conjugate according to the present invention is superior to the enhancement effect of the anti-tumor effect when an immunostimulatory agent is conjugated to an existing cancer antibody drug. Therefore, the conjugate according to the present invention is effective for the treatment or prevention of cancer.
MODE FOR CARRYING OUT THE INVENTION
[0012]
　The antitumor activity of the conjugate of an antibody against a CAPRIN-1 protein (hereinafter referred to as "anti-CAPRIN-1 antibody") used in the present invention and an immunostimulatory factor is carried in vivo as described later. It can be evaluated by examining suppression of tumor growth on cancer animals.
[0013]
　In the present invention, "conjugate" refers to one in which an antibody and an immunostimulatory agent are covalently linked. A linker may be used to bind the antibody to the immunostimulatory factor.
[0014]
　The anti-CAPRIN-1 antibody forming the above-mentioned conjugate according to the present invention may be a monoclonal antibody or a polyclonal antibody, preferably a monoclonal antibody, as long as the conjugate of the present invention can exert antitumor activity. The antibody may be any type of antibody, and the antibody is a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, a non-human animal antibody.
[0015]
　The immunostimulatory agent forming the above-mentioned conjugate according to the present invention may be a factor which activates immune cells, preferably Toll-like receptor (TLR), NOD-like receptor (NLR), RIG-like It is a receptor, a ligand that binds to the C-type lectin receptor (CLR), an agonist or a derivative thereof. More preferably, they are ligands or agonists that bind to Toll-like receptor (TLR) and derivatives thereof.
[0016]
　Further, the subject to be treated and / or prevented for cancer in the present invention is a mammal such as human, pet animals, livestock animals and sports animals, and a preferred subject is a human.
[0017]
　The following describes the anti-CAPRIN-1 antibody, the immunostimulatory factor, the conjugate of the anti-CAPRIN-1 antibody and the immunostimulatory factor, the pharmaceutical composition using the conjugate and the method for treating and / or preventing cancer according to the present invention Do.
[0018]
　
　A CAPRIN- having an amino acid sequence represented by any one of SEQ ID NOs: 2 to 30, which has immunological reactivity with the anti-CAPRIN-1 antibody used in the present invention. Among the proteins, the amino acid sequences shown in SEQ ID NOs: 6, 8, 10, 12 and 14 are amino acid sequences of canine CAPRIN-1 protein, and the amino acid sequences shown in SEQ ID NOs: 2 and 4 are human The amino acid sequence of the protein, the amino acid sequence shown in SEQ ID NO: 16 is the amino acid sequence of the bovine CAPRIN-1 protein, the amino acid sequence shown in SEQ ID NO: 18 is the amino acid sequence of the equine CAPRIN-1 protein, The amino acid sequences shown in SEQ ID NOS: 20-28 are of the mouse CAPRIN-1 protein An amino acid sequence, the amino acid sequence shown in SEQ ID NO: 30 is the amino acid sequence of caprin-1 protein of chicken.
[0019]
　In addition, the anti-CAPRIN-1 antibody used in the present invention has an amino acid sequence represented by any one of SEQ ID NOs: 2 to 30 and an even numbered SEQ ID NO: 80% or more, It may have immunological reactivity with a variant of CAPRIN-1 protein having% or more, more preferably 99% or more of sequence identity. As used herein, “% sequence identity” refers to the amino acid (or base) when two sequences are aligned (aligned) with maximum similarity, with or without introducing gaps. It means the percentage (%) of the same amino acid (or base) to the total number.
[0020]
　The anti-CAPRIN-1 antibody used to produce a conjugate in the present invention means an antibody or a fragment thereof having immunological reactivity with the full length of CAPRIN-1 protein or a fragment thereof. Here, "immunological reactivity" refers to the property of an antibody binding to CAPRIN-1 protein or a partial polypeptide thereof in vivo.
[0021]
　The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.
[0022]
　A polyclonal antibody (anti-CAPRIN-1 polyclonal antibody) having immunological reactivity with the full length or a fragment thereof of CAPRIN-1 protein is, for example, a natural CAPRIN-1 protein, a fusion protein with GST or the like, or a partial peptide thereof The mice, human antibody-producing mice, rats, rabbits, chickens, etc. are immunized to obtain serum. The obtained serum can be obtained by ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, affinity column to which CAPRIN-1 protein or partial peptide is bound, or the like.
[0023]
　The full-length or fragment thereof of CAPRIN-1 protein used for the above-mentioned immunity, the base sequence and amino acid sequence of CAPRIN-1 and its homologs are accessed, for example, GenBank (US NCBI), and algorithms such as BLAST, FASTA etc (Karlin and Altschul , Proc. Natl. Acad. Sci. USA, 90: 5873-5877, 1993; Altschul et al., Nucleic Acids Res. 25: 3389-3402, 1997). In addition, the method for producing the CAPRIN-1 protein can be obtained by referring to WO 2014/012479, and cells etc. expressing the CAPRIN-1 protein can also be used.
[0024]
　A monoclonal antibody (anti-CAPRIN-1 monoclonal antibody) having immunological reactivity with the full length or a fragment thereof of CAPRIN-1 protein is, for example, a breast cancer cell SK-BR-3 or CAPRIN-1 protein that expresses CAPRIN-1. A full length or a fragment thereof or the like is administered to a mouse to immunize, spleen cells and myeloma cells isolated from the same mouse are fused, and a clone producing an anti-CAPRIN-1 monoclonal antibody is selected from the resulting fused cells (hybridoma). It can be obtained by An antibody produced from a selected hybridoma can be obtained by the same method as the method for purifying a polyclonal antibody described above.
[0025]
　Antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies and non-human animal antibodies.
[0026]
　A human antibody is prepared by sensitizing human lymphocytes infected with EB virus with a protein, a protein expressing cell or a lysate thereof, and fusing the sensitized lymphocytes with myeloma cells such as human-derived U266 cells, and the resulting fusion An antibody having immunological reactivity with the full length of CAPRIN-1 protein or a fragment thereof can be obtained from cells.
[0027]
　Humanized antibodies are modified antibodies, also referred to as reshaped human antibodies. Humanized antibodies are constructed by implanting the complementarity determining regions of antibodies derived from the immunized animal into the complementarity determining regions of human antibodies. The gene recombination method as the general method is also a well-known technique. Specifically, for example, several DNA sequences designed to link the complementarity determining region of mouse antibody and rabbit antibody with the framework region of human antibody were prepared so as to have an overlapping portion at the end. Are synthesized by PCR from the oligonucleotides of The obtained DNA is ligated with the DNA encoding the constant region of a human antibody, incorporated into an expression vector, and introduced into a host for production (European Patent Application Publication No. EP 239 400, International Publication Number See WO 96/02576). The framework regions of the human antibody linked via the complementarity determining regions are selected such that the complementarity determining regions form a good antigen binding site. If necessary, the amino acid of the framework region in the variable region of the antibody may be substituted so that the complementarity determining region of the reshaped human antibody forms a suitable antigen binding site (Sato K. et al., Cancer Research 1993, 53: 851-856). Also, framework regions derived from various human antibodies may be substituted (see WO 99/51743).
[0028]
　An antibody is a heteromultimeric glycoprotein, usually comprising at least two heavy chains and two light chains. An antibody is composed of two identical light chains and two identical heavy chains. The heavy chain has at one end a heavy chain variable region followed by a number of constant regions. The light chain has at one end a light chain variable region followed by a number of constant regions. The variable regions indicate specific variable regions called complementarity determining regions (CDRs) to impart binding specificity to the antibody. The portion relatively stored in the variable region is called a framework region (FR). The variable regions of the complete heavy and light chains comprise four FRs linked by three CDRs (CDR1-CDR3), respectively.
[0029]
　The sequences of constant regions and variable regions of human-derived heavy and light chains can be obtained, for example, from NCBI (US: GenBank, UniGene, etc.). For example, the heavy chain constant region of human IgG1 has the accession number J00228, For the heavy chain constant region of human IgG2, refer to the sequence such as accession number J00230, for human light chain 定 常 constant region accession number V00557, X64135, X64133 etc. For the human light chain λ constant region, accession number be able to.
[0030]
　A chimeric antibody is an antibody produced by combining sequences derived from different animals, and for example, from the constant regions of heavy chain variable region and light chain variable region of mouse antibody and heavy chain variable region and light chain variable region of human antibody And the like. A chimeric antibody can be produced using a known method, for example, a DNA encoding the antibody V region and a DNA encoding the C region of a human antibody are linked, and this is incorporated into an expression vector and introduced into a host It is obtained by producing.
[0031]
　The non-human animal antibody can be obtained by immunizing an animal with a sensitizing antigen according to a known method. As a general method, the sensitizing antigen is obtained by intraperitoneal, intradermal or subcutaneous injection of an animal such as a mouse. . When injecting a sensitizing antigen, it is mixed with an appropriate amount of various adjuvants such as CFA (Freund's complete adjuvant) and administered to animals multiple times. After immunizing the animal and confirming that the serum contains anti-CAPRIN-1 antibody, serum is obtained, and as described above, ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, CAPRIN-1 protein And an affinity column to which a partial peptide is bound. In addition, when monoclonal antibodies are obtained from non-human animals, they can be obtained by collecting immune cells from immunized animals and subjecting them to cell fusion with myeloma cells. The cell fusion between the immune cells and the myeloma cells can be performed according to known methods (see Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).
[0032]
　The antibody used in the present invention may also be obtained as a recombinant antibody produced by cloning an antibody gene from a hybridoma, incorporating it into a suitable vector, and introducing it into a host using gene recombination technology. (See Carl, A. K. Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990).
[0033]
　The anti-CAPRIN-1 antibody used to obtain the conjugate of the present invention may be one in which an amino acid in the variable region (eg, FR) or the constant region is substituted with another amino acid. The amino acid substitution is a substitution of one or more, for example, less than 15, 10, 8 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less amino acids, preferably 1 to 9 amino acids. The antibody should have the property of binding specifically to the antigen, the affinity for binding to the antigen equal to or higher than that of the unsubstituted antibody, and not cause rejection upon application to human beings .
[0034]
　The anti-CAPRIN-1 antibody used in the present invention can be expected to have a stronger antitumor effect as the binding affinity to the CAPRIN-1 protein on the surface of cancer cells is higher. The binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5 × 10 8 M -1 , at least 10 9 M -1 , at least 5 × 10 9 M −1 , at least 10 10 M −1 , at least 5 × 10 10 M −1 , at least 10 11 M −1 , at least 5 × 10 11 M −1 , at least 10 12M -1 or at least 10 13 M -1 is desirably.
[0035]
　The anti-CAPRIN-1 antibody used in the present invention substitutes one, two or several amino acids in the heavy chain constant region of the antibody, or N-glycoside linked N-glycoside linked to the heavy chain constant region By removing fucose bound to acetylglucosamine, the binding ability of anti-CAPRIN-1 antibody to effector cells can be improved. The above may be amino acid substitution alone, or may be a composition with an antibody to which fucose is bound.
[0036]
　Antibodies in which one, two or several amino acids in the heavy chain constant region have been substituted are prepared, for example, with reference to WO 2004/063351, WO 2011/120135, US Patent 8388955, WO 2011/050481, US Patent 6373056, WO 2005/063351. can do.
[0037]
　An antibody from which fucose attached to N-acetylglucosamine in N-glycosidic linked sugar chain in heavy chain constant region has been removed or a cell producing the same is referred to US Pat. No. 6,602,684, European patent 1914244, US Pat. 7579170 Can be produced. A composition of an antibody in which fucose is removed and an antibody from which fucose is bound to N-acetylglucosamine in an N-glycoside-linked sugar chain bound to a heavy chain constant region or a cell producing the same is described in, eg, US Pat. Can be made with reference to the
[0038]
　The anti-CAPRIN-1 polyclonal antibody, anti-CAPRIN-1 monoclonal antibody, antibody production method, purification method and CAPRIN-1 protein or partial polypeptide thereof used for immunization according to the present invention are disclosed in WO 2010/016526, WO 2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 125,640, it is possible to WO2013 / 147169, obtained by reference to WO2013 / 147176 and WO2015 / 020212.
[0039]
　Specific examples of the anti-CAPRIN-1 antibody in the present invention include the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 Anti-CAPRIN as described in: -1 antibody is mentioned As the preferred anti-caprin-1 antibody include the following.
[0040]
　80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, still more preferably 99% or more) with the amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4 An antibody or a fragment thereof having immunological reactivity with a partial polypeptide of CAPRIN-1 protein having an amino acid sequence having the sequence identity of
[0041]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 31 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 36, 37 and 38 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 40, 41 and 42 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein, or the complementarity determining regions of SEQ ID NO: 140, 141 and 142 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 143, 144 and 145, respectively), and having immunological reactivity with the CAPRIN-1 protein Antibody or fragment thereof, or SEQ ID NO: 164, 165 and 1 A heavy chain variable region comprising six complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of SEQ ID NOs: 167, 168 and 169 And an antibody or fragment thereof having immunological reactivity with CAPRIN-1 protein. More preferably, the antibody or fragment thereof in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43, or the heavy chain variable region comprises SEQ ID NO: 70 Or a fragment thereof, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71, or the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 79.
[0042]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 33 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable region containing the complementarity determining regions of SEQ ID NO: 60, 61 and 62 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 64, 65 and 66 (CDR1, CDR2 and CDR3 respectively) And an antibody or fragment thereof immunologically reactive with a CAPRIN-1 protein, or the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63, and a light chain variable An antibody or fragment thereof, wherein the region comprises the amino acid sequence of SEQ ID NO: 67.
[0043]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 32 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable region containing the complementarity determining regions of SEQ ID NO: 52, 53 and 54 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 56, 57 and 58 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region comprising: and having immunological reactivity with a CAPRIN-1 protein. More preferably, an antibody or fragment thereof wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59.
[0044]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 34 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 170, 171 and 172 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 173, 174 and 175 (CDR1, CDR2 and CDR3 respectively) Antibodies or fragments thereof having immunological reactivity with a CAPRIN-1 protein, or the complementarity-determining regions of SEQ ID NO: 176, 177 and 178 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 179, 180 and 181 (CDR1, CDR2 and CDR3 respectively) and immunologically reactive with the CAPRIN-1 protein An antibody or fragment thereof. More preferably, the antibody or fragment thereof in which 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, or the heavy chain variable region comprises SEQ ID NO: 82 And an amino acid sequence of SEQ ID NO: 83, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83.
[0045]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 35 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 182, 183 and 184 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 185, 186 and 187 (CDR1, CDR2 and CDR3 respectively) Antibodies or fragments thereof having immunological reactivity with a CAPRIN-1 protein, or the complementarity-determining regions of SEQ ID NO: 188, 189 and 190 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 191, 192 and 193, respectively) and immunologically reactive with the CAPRIN-1 protein An antibody or fragment thereof. More preferably, the antibody or fragment thereof in which 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, or the heavy chain variable region comprises SEQ ID NO: 86 And an amino acid sequence of SEQ ID NO: 87, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87.
[0046]
　A light chain comprising a heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of SEQ ID NO: 44 and 45 and a complementarity determining region (CDR1, CDR2 and CDR3 respectively) of SEQ ID NO: 48, 49 and 50 An antibody or fragment thereof comprising a chain variable region and having immunological reactivity with a CAPRIN-1 protein. Preferably, an antibody or fragment thereof wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 47 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 51.
[0047]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 296 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 146, 147 and 148 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 149, 150 and 151 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0048]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 297 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 272, 273 and 274 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 275, 276 and 277 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0049]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 298 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 290, 291 and 292 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 293, 294 and 295 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0050]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 299 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 301, 302 and 303 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 305, 306 and 307 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0051]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 308 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 134, 135 and 136 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 137, 138 and 139 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0052]
　CAPRIN-having an amino acid sequence represented by SEQ ID NO: 309 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) An antibody or fragment thereof having immunological reactivity with a partial polypeptide of 1 protein. Preferably, the heavy chain variable regions comprising the complementarity determining regions of SEQ ID NO: 134, 135 and 136 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 137, 138 and 139 (CDR1, CDR2 and CDR3 respectively) An antibody or fragment thereof comprising a light chain variable region and having immunological reactivity with a CAPRIN-1 protein. More preferably, 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.
[0053]
　Also, the following anti-CAPRIN-1 antibodies are preferably used.
[0054]
　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.
[0055]
　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.
[0056]
　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.
[0057]
　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.
[0058]
　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.
[0059]
　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.
[0060]
　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.
[0061]
　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.
[0062]
　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.
[0063]
　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.
[0064]
　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.
[0065]
　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.
[0066]
　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.
[0067]
　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.
[0068]
　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.
[0069]
　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.
[0070]
　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.
[0071]
　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.
[0072]
　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.
[0073]
　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.
[0074]
　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.
[0075]
　An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111.
[0076]
　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.
[0077]
　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.
[0078]
　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.
[0079]
　An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119.
[0080]
　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.
[0081]
　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.
[0082]
　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.
[0083]
　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.
[0084]
　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.
[0085]
　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.
[0086]
　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.
[0087]
　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.
[0088]
　In the following example, a conjugate with an immunostimulatory agent is prepared using the above-described polyclonal antibody and monoclonal antibody against a full-length CAPRIN-1 protein, a part of the polypeptide expressed in the cell membrane surface of cancer cells, The strong antitumor effect was confirmed.
[0089]
　
　The immune activator according to the present invention is a factor that activates various immune cells, and means a natural compound, nucleic acid or natural compound that can maintain and enhance the immune function of the cells. . The term "immune cells" as used herein refers to T lymphocytes, B lymphocytes, NK cells, monocytes, dendritic cells, granulocytes, macrophages, bone marrow-derived suppressor cells, Langerhance cells and their precursor cells, and tumors. It is the said immune cell group which exists.
[0090]
　The immunostimulatory factor used in the present invention is not particularly limited. Specific examples thereof include Toll-like receptor (TLR), NOD-like receptor (NLR), RIG-like receptor or C-type lectin receptor (CLR) And ligands or agonists that bind to
[0091]
　Specific examples of ligands or agonists that bind to Toll-like receptor (TLR) include ligands or agonists that bind to TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, TLR10.
[0092]
　Specific examples of the ligand or agonist binding to TLR2 include substances selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide and zymosan.
[0093]
　Specific examples of the ligand or agonist that binds to TLR3 include substances selected from the group consisting of Poly (I: C) and poly (A: U), polyICLC (Hiltonol) and Ampligen.
[0094]
　Specific examples of the ligand or agonist binding to TLR4 include lipopolysaccharide (LPS), HSP60, RS09, a substance selected from the group consisting of MPLA (Monophosphoryl lipid A from Salmonella minnesota R595), GLA-SE, G100 and MPLA. Can be mentioned.
[0095]
　Specific examples of ligands or agonists that bind to TLR5 include substances selected from the group consisting of flagellin and FLA.
[0096]
　Specific examples of ligands or agonists that bind to TLR7 or TLR8 include low molecular weight compounds such as imidazoquinolines compounds, single-stranded RNAs, etc. Specific examples thereof include Imiquimod, Resiquimod, Loxorbine, 852A, 854A, S-34240, Motolimod (VTX-2337), DSR-6434, GS-9620, ANA773, AZD8848 / DSP-3025, GSK2245035, Gardiquimod, CL264, UC-1V150, CL075, CL097, CL307, CL347, 3M-003, 3M -0043, 3M-052, CL264, IV209, ORN R-2176-dT, Poly (dT), ORN R-0006, ORN R-0002, ORN R-2336, PolyU, ORN R-1886, polyG3, DSR6434, RWJ21757, SM324405, p-IMDQ, m-IMDQ and GSK2245035.
[0097]
　Specific examples of ligands or agonists that bind to TLR9 include bacterial DNA, unmethylated CpG DNA, hemozorin, ODN 1585, ODN 1668, ODN 1668, Lefitolimod (MGN1703), SD-101, CYT003, CPG7909, DUK-CPG-001 and ODN1826 And a substance selected from the group consisting of
[0098]
　Specific examples of ligands or agonists that bind to TLR10 include substances selected from the group consisting of profilin and uropathogenic bacteria.
[0099]
　Among ligands or agonists that bind to the Toll-like receptor (TLR), in the present invention, ligands or agonists that bind to TLR7 or TLR8 are preferably used, and as ligands or agonists that bind to TLR7 or TLR8, imidazoquinolines compounds and More preferably, a ligand or agonist that binds to TRL7 or 8 selected from the group consisting of single stranded RNA is used, and more preferably a ligand or agonist that binds to TRL7 or 8 selected from imidazoquinolines compounds Be
[0100]
　Preferred specific examples of the imidazoquinolines compounds include the compounds described in US Pat. No. 8,951,528 and the compounds described in WO2015 / 103989, for example, 1- (2-methylpropyl) -1H. -Imidazo [4,5-c] quinoline-4-amine (Imiquimod), 1- (4-amino-2-ethylaminomethylimidazo- [4,5-c] quinolin-1-yl) -2-methylpropan-2-ol (Gardiquimod), N- [4- (4-amino-2-ethyl-1H-imidazo [4,5-c] quinolin-1-yl) butyl-] methanesulfonamide (PF-4878691), 4-a mino-2- (ethoxymethyl) -a, a-dimethyl-1H-imidazo [4,5-c] quinoline-1-ethanol (Resiquimod), 4-amino-aa-dimethyl-2-methoxyethyl-1H-imidazo [4 , 5-c] quinoline-1-ethanol, 1- (2- (3- (benzyloxy) propoxy) ethyl) -2- (ethoxymethyl) -1H-imidazo [4, 5-c] quinolin-4-amine, 4 -Amino-2-ethoxymethyl-aa-dimethyl-6,7,8,9-tetrahydro-1H-imidazo [4,5-c] quinoline-1-ethanol, N- (2- { -[4-amino-2- (2-methoxyethyl) -1H-imidazo [4,5-c] quinolin-l-yl] eyhoxy} ethyl) -n'-phenylurea, 1-2-amino-2-methylpropyl) -2- (ethoxymethyl) -1H-imidazo [4,5-c] quinolin-4-amine, 1- {4-[(3,5-dichloro) phenyl) sulfonyl] butyl} -2-ethyl-1H-imidazo [4,5-c] quinolin-4-amine, N- (2- {2- [4-amino-2- (ethoxymethyl) -1H-imidazo [ 4,5-c] quinolin-1-yl] ethoxy} ethyl) -n'-cyclohexylurea, N- {3- [4-amino-2- (ethoxymethyl) -1H-imidazo [4,5-c] quinolin- 1-yl] propyl} -n '-(3-cyanophenyl) thiourea, N- [3- (4-amino-2-butyl-1H-imidazo [4,5-c] quinolin-l-yl) -2, 2-dimethylpropyl] benza ide, 2-butyl-1- [3- (methylsulfonyl) propyl] -1H-imidazo [4,5-c] quinolin-4-amine, but is not particularly limited as long as it binds to TLR7 or TLR8 .
[0101]
　Specific examples of ligands or agonists that bind to NOD-like receptor (NLR) include, for example, M-TriDAP, PGN, and others, which are ligands or agonists for NOD1, such as Tri-DAP, iE-DAP, C12-iE is mentioned. Moreover, it is a ligand or agonist for NOD2 and includes MDP, N-glycosyl-MDP, Murabutide, M-TriLyS-D-ASN, M-TriLYS, L18-MDP.
[0102]
　Specific examples of ligands or agonists that bind to RIG-like receptors include, for example, 5 'ppp-dsRNA, poly (dA: dT), poly (dG: dC), poly (I: C) .
[0103]
　Specific examples of ligands or agonists that bind to the C-type lectin receptor (CLR) include trehalose 6,6-dibehenate, zymosan, WGP, HKSC, HKCA and curdlan AL.
[0104]
　 In the
　present invention, the binding form of the anti-CAPRIN-1 antibody and immunostimulatory agent in the conjugate of anti-CAPRIN-1 antibody and immunostimulatory factor is The form is not particularly limited as long as it can maintain the anti-tumor activity against B. However, it is preferable that the form is a bond in which a linker structure is formed between the anti-CAPRIN-1 antibody and the immunostimulatory agent.
[0105]
　Here, a linker means a compound capable of binding an anti-CAPRIN-1 antibody and an immunostimulatory factor. Various known linkers may be used, or the structure on the side of the activator may be subjected to appropriate chemical modification and directly linked.
[0106]
　The type of linker and details of the binding method can be performed according to known methods (see, for example, Greg T. Hermanson Bioconjugate Techniques, Third Edition and WO 2004010957, WO 2014/012479).
[0107]
　In embodiments of the invention, reactive groups attached to anti-CAPRIN-1 antibodies, immunostimulatory factors and linkers include:
[0108]
　As the reactive group attached to the amino acid sequence of the antibody or the glycoprotein modified to the amino acid, unless special chemical modification is made, primary amine (ε-amino group), carboxyl, thiol (sulfhydryl), Included are carbonyl (ketone or aldehyde) as well as hydroxyl. A primary amine is present at the N-terminus of the polypeptide or at the side chain of a lysine residue, is positively charged under physiological conditions, and is usually outside the protein, thus altering the structure of the protein It can be used without binding. The carboxyl is present at the C-terminus of the polypeptide and at the side chains of aspartic acid and glutamic acid. Sulfhydryl is present in the side chain of cysteine ​​and forms a disulfide bond that maintains the conformation of the protein. Ketone or aldehyde groups are made in glycoproteins by oxidizing glycosyl with sodium metaperiodate.
[0109]
　The conjugate of the present invention is prepared by attaching a linker, attaching a linker, attaching an immunostimulatory agent to which a linker is attached, or directly attaching an immunostimulatory agent to an antibody.
[0110]
　The linker and the reactive group attached to the immunostimulatory factor include the following.
[0111]
　N-hydroxysuccinimide (NHS) ester, imido ester, pentafluorophenyl ester, hydroxymethyl phosphine, isothiocyanate, isocyanate, acyl azide, N-hydroxy ester, sulfonyl chloride, aldehyde, glyoxal, epoxide which is a reactive group capable of reacting to amine , Oxiranes, carbonates, aryls, imidoesters, carbodiimides and carboxylic acid anhydrides.
[0112]
　Carbodiimides, diazoalkanes, diazoacetyl compounds, carbonyldiimidazole which are reactive groups capable of reacting to carboxyl and amine.
[0113]
　Reactive groups capable of reacting to thiol, maleimide, haloacetamide, pyridyl disulfide, thiosulfone, vinyl sulfone, haloacetyl, aziridine, acryloyl, aryl.
[0114]
　Hydrazides and alkoxyamines which are reactive groups capable of reacting to aldehydes. Epoxy, oxirane, carbonyldiimidazole, N, N'-disucciimidyl carbonate, N-hydroxysuccinimidyl chloroformate, isocyanate which is a reactive group capable of reacting to hydroxyl.
[0115]
　Isocyanate which is a reactive group capable of reacting to hydroxyl.
[0116]
　Diazirine, aryl azide, aryl, benzophenol, diazo compound as a photoreactive reactive group.
[0117]
　Specific examples of the linker having the above reactive group include the following.
[0118]
　As a linker having the same reactive group end, a linker having N-hydroxysuccinimide ester as a reactive group (for example, 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), (DTSSP), (EGS), (Sulfo-EGS), (Sulfo-EGS), DMA), (DMP), (DMS), (D) BP), (DFDNB), (DST), (BSOCOES), (EGS), (Sulfo-EGS)), and a linker having maleimide as a reactive group (eg, (BMOE), (BMB), (BMH), (TMEA) (BM (PEG) 2), (BM (PEG) 3 ), (DTME), (DMDB)) are used.
[0119]
　As main linkers having different reactive group ends, linkers using NHS ester and maleimide as reactive groups (eg, 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), Linkers having NHS ester and pyridyldithiol as reactive groups (eg SPDP, LC-SPDP, Sulfo-LC-SPDP, SMPT, PEG4 horizontal SPDP, PEG12-SPDP), linkers having NHS ester and haloacetyl as reactive groups ( For example, SIA, SBAP, SIAB, Sulfo-SIAB), a linker having NHS ester and aryl azide as a reactive group (eg, ANB-NOS, Sulfo-SANPAH, ATFB), a linker having NHS ester and diazirine as a reactive group (eg, SDA, Sulfo-SDA, LC-SDA, SDAD, Sulfo-SDAD), carbodiimide-based linker (eg DCC, EDC, EDAC, NHS, Sulfo-NHS), maleimide and hydrazide with reactive groups Linkers (eg BMPH, EMCH, MPBH, KMUH), linkers with pyridyldithiol and hydrazide as reactive groups (eg PDPH), linkers with isocyanate and maleimide as reactive groups (eg PMPI) and NHS esters with psoralens as reactive groups A linker (eg SPB) is used.
[0120]
　As other linkers, linkers comprising polypeptides, such as Fmoc-Ala-Ala-Asn-PAB, Fmoc-Ala-Ala-Asn (Trt) -PAB, 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 -Asn -Ala (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.
[0121]
　In addition, Bis-PEG-acid, PEG Acid (eg, Acid-PEG-TEMPO, Amino-PEG-acid, Amino-PEG-CH 2 CO 2 H, Aminoxy-PEG-acid, Azido-PEG-acid, Carboxy-PEG- Sulfonic acid, Fmoc-N-amido-PEG-acid, Fmoc-N-amido-PEG-CH 2 CO 2 H, Fmoc-aminooxy-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, Propar -PEG-yl acid, propargyl-PEG-CH 2 CO 2 H, propargyl-PEG-(CH 2 ) 3 -Acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG- CH 2CO 2 H, t-Boc-Aminooxy-PEG-acid, Acid-PEG-PFP ester, Miscellaneous PEG acid,), PEG PFP ester (eg, Acid-PEG-PFP ester, Bis-PEG-PFP ester), Bis- PEG-NHS, PEG Aldehyde (eg, 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 (eg Azido-PEG-Tos, Hy roxy-PEG-Tos, m-PEG-Tos, t-Boc-Aminooxy-PEG-Tos, Trifluoroethyl-PEG-Tos, Tos-PEG-acid, Tos-PEG-CH 2 CO 2 H, Tos-PEG-Alkyne, Tos-PEG-t-butyl ester, Tos-PEG-CH 2 CO 2 tBu, Tos-PEG-Tos, S-acetyl-PEG6-Tos, N-Tos-N- (t-butoxycarbonyl) -aminooxy-PEG 4-Tos, Ms-PEG-Ms, Ms-PEG-t-butyl ester, PEG-Ms, Propargyl-PEG-Ms), Boc-PEG (eg, 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-bromode, N-methyl-N- (t-Boc) -PEG-acid, t-Boc-N-amido-PEG-acid, t-Boc-N-amido-PEG-CH 2 CO 2 H, t -Boc-N-Amid -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 (eg, 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 (eg Fmoc-N-) -PEG-mido acid, Fmoc-NH-PEG-CH 2 CO 2H, Fmoc-PEG-NHS ester), Biotin PEG (eg Biotin PEG-acid, Biotin PEG-alcohol, Biotin PEG-alkyne, Biotin PEG-amine, Biotin PEG-azide, Biotin PEG-DBCO, Biotin PEG-hydrazide, Biotin -PEG-Mal, Biotin-PEG-NHS, Biotin-EDA-PEG-NHS, Biotin-PEG-oxyamine, Biotin-PEG-PFP, Biotin-EDA-PEG-PFP, Biotin-PEG-Tetrazine, Biotin-PEG-TFP , Azide-SS-biotin, Biotin-PEG3-SS-azide, DBC -S--S PEG3-Biotin, Dde Biotin-PEG4-Alkyne, Dde Biotin-PEG 4 -Azide, Dde Biotin-PEG 4 -DBCO, Diazo Biotin-PEG 3 -Alkyne, Diazo Biotin-PEG 3 -Azide, Diazo Biotin DBCO--PEG3, 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-PEG4-PEG3-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 (eg 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-Bramide, 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 (eg, endo-BCN-PEG, exo-BCN-PEG, Propargyl-PEG-ac id, Propargyl-PEG-CH 2 CO 2 H, Propargyl-PEG- (CH 2 ) 3-Acid, Propargyl-PEG- (CH 2 ) 3 -methyl ester, Propargyl-PEG-Acrylate, Propargyl-PEG-alcohol, Propargyl-PEG-amine, Propargyl-PEG-methylamine, Aminooxy-PEG-Propargyl, Propargyl-PEG- Azide, Propargyl-PEG-Bromide, Propargyl-PEG-Maleimide, Propargyl-PEG-Ms, Propargyl-PEG-NHS ester, Propargyl-PEG-sulfonic acid, Propargyl-PEG-t-butyl ester, Propargyl-PEG-CH 2 CO 2 tBu, Propargyl-PEG-thiol, Propargyl-PEG-5-nitrophenyl carbonate, t-Boc-aminooxy-PEG-Propargyl, Bis-Propargyl-PEG, m-PEG-Propargyl), Azido PEG (eg, Azido-PEG) -Acid, Azido-PEG-CH 2 CO 2 H, Azido-PEG- (CH 2 ) 3-Methyl ester, Azido-PEG-Acrylate, Azido-PEG-alcohol, Azido-PEG- (CH 2 ) 3 OH, Azido-PEG-amine, Azido-PEG-azide, Azido-PEG-Maleimide, Azido-PEG-methylamine Azido-PEG-methyl ester Azide-PEG-NHS ester Azido-PEG-CH 2 CO 2 -NHS Azido-PEG-oxazolidin-2-one Azido-PEG-PFP ester Azido-PEG-phosphonic acid Azido-PEG-phosphonic acid ethyl ester, Azido-PEG-sulfoni c 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, Aminooxy-PEG-azide, Bromo-PEG-azide, Bromoacetamido-PEG-azide, Carboxyrhodamine 110-PEG-Azide, Isothiocyanato-PEG-Azide, Isothiocyanato-PEG-Azide, m-PEG-Azide, Propargyl-PEG-Azide, TAMRA-PEG-Azide, t-Boc-N-Amido-PEG-Azide, t-Boc-Aminooxy-PEG-Azide, Thiol-PEG- Azide, Trifluoroethyl-PEG-Azide, Azido-P -Amino acid G, 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 are used.
[0122]
　The linker between the anti-CAPRIN-1 antibody and the immunostimulatory factor may be composed of a single species or multiple species.
[0123]
　As a method of producing a conjugate of the anti-CAPRIN-1 antibody of the present invention and an immunostimulatory factor, a method of binding an immunostimulatory factor by using the ε-amino group of the lysine side chain of the antibody, disulfide of the antibody There is a method of binding a cysteine ​​residue forming a bond using a thiol formed by reduction treatment.
[0124]
　When the ε-amino group of a lysine residue of an antibody is used, for example, a method of reacting an active ester (eg, N-hydroxysuccinimide ester) to form an amide bond is used. In this case, the binding proceeds nonspecifically because the antibody has many lysine residues.
[0125]
　When using the thiol which forms the disulfide bond which exists in the side chain of the cysteine ​​of an antibody, the disulfide bond which exists on an antibody is made to generate thiol using reducing agents, such as mercaptoethanol, and maleimide or alpha-haloamide The method of making it react is used. In addition, in order to stabilize the bond via thiol, for example, there is a method using sulfone phenyl oxadiazole, 4-cyanoethynyl oxy derivative or the like. These bonds are stable for a longer period of time than the conjugation of cysteine ​​to maleimide by conjugation. In addition, since the imide ring in which the thiol group is added to maleimide is opened by hydrolysis to form an amido bond and the stability is improved, a linker having an amino group in the vicinity of the imide group can also be used. In addition, the thiol of cysteine ​​in the antibody forms a disulfide bond, and there is also a method of binding an immunostimulatory factor or the like between them via two thiols. As an example, it is possible to form a crosslinkable bond using a linker with two disulfide bonding sites or dibromomaleimide which can be generated from an amide group having two sulfones in the β position.
[0126]
　Conjugates of the present invention, for example, to a particular portion of the antibody, ThioMab is a method capable of introducing fixed number of thiol groups TM is a method using a technique (see Nature Biotechnology 26,925-932 (2008)) .
[0127]
　The conjugate of the present invention is conjugated with an immunostimulatory agent, for example, by obtaining an antibody having a thiol reactive group by reducing the antibody using a reducing agent dithioureitol (DTT) in a phosphate buffer. Can be formed. In addition to the method using a reducing agent, a thiol group is added to the primary amine at the lysine residue of the antibody by introducing a trout reagent (2-Iminothiolane or N-Succinimidyl S-Acetylthioacetate (SATA)). You can get it.
[0128]
　The amount of thiol added to the antibody can be determined, for example, by mixing 5,5'-Dithiobis (2-nitrobenzoic acid) (DTNB) and a sample solution containing an SH group with phosphate buffer (pH 8.0) and distilled water, It can be quantified by measuring the absorbance at 412 nm after adding a DTNB solution dissolved in an acid buffer solution, a Good buffer solution or a Tris buffer and incubating for a certain period of time (GL Ellman, Arch. Biochem. Biophys., 82, 70 (1959)).
[0129]
　The thiol group added by cleaving the disulfide bond of the antibody by reduction treatment is preferably subjected to treatment (capping) to prevent formation of the disulfide bond again. For capping, for example, N-ethyl maleimide (NEM) or iodoacetamide (2-iodoacetamide (IAA)) can be used.
[0130]
　A known method can be used to bind an immunostimulatory agent to form a conjugate using a thiol group added to an antibody. Specifically, for example, as a linker reagent that specifically binds to a thiol group of a reduction-treated antibody, one having a maleimide group or a linker reagent having a bromoacetamide group can be used. For example, N-succinimidyl-4- (N-maleimidomethyl) -cyclohexane-1-carboxylate (SMCC) is used as a linker having a maleimide group. In this case, a conjugate can be obtained by forming an amide bond with the N-succinimide group of SMCC due to the presence of an amino group in the immune activation factor.
[0131]
　In another embodiment, SMCC is used to form an amide bond to the amino group previously present in the activating factor, and then the thiol group added to the antibody side and the maleimide group of SMCC to which the immunostimulatory factor is bound are used. The reaction can be performed to obtain a conjugate.
[0132]
　In another embodiment, it is also possible to form a conjugate between the antibody and the immunostimulatory agent by using two linkers. For example, a thiol group is prepared by preparing a product in which a primary amino group present in a lysine residue on the antibody side and an N-succinimide group of SATA (N-succinimidyl-S-acetylthioacetate) are bound to form an amide bond. Addition is carried out, and those having an amino group on the side of the immunostimulatory factor or those having an amino group added thereto are reacted with SMCC to form an N-succinimide group and an amide bond in SMCC, A conjugate can be obtained by the reaction of a maleimide group in SMCC to which an immunostimulatory factor is bound and a thiol group on SATA to which an antibody is bound.
[0133]
　In another embodiment, a method using, for example, maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-Val-Cit-PAB) as a linker is used for producing a conjugate of an antibody and an immunostimulatory factor Can be mentioned. MC-val-Cit-PAB is a linker cleavable by an intracellular protease (eg cathepsin B). A thiol group is added to the antibody dissolved in phosphate buffer using DTT or the like. On the other hand, an immune activation factor having an amino group is reacted with benzyloxycarbonyl (PAB) in MC-Val-Cit-PAB to produce an immune activation factor bound with MC-val-Cit-PAB. The antibody can be reacted with a thiol-added antibody to obtain a conjugate.
[0134]
　In yet another embodiment, SATA is attached to a primary amino group at a lysine residue of an antibody to add a thiol group, while succinimidyl 3- (2-pyridyldithio) propionate (SPDP) is added to an amino group. In
order to obtain a composition containing an antibody in which an N-succinimide group in SPDP and an amide bond in the SPDP are bound and reacted with an immune activating factor having the antibody, the linker is subjected to, for example, gel filtration chromatography. It can be obtained by fractionating a more polymerized peak as compared to the antibody before binding.
[0135]
　The number of immunostimulatory agents bound per antibody of the conjugate of anti-CAPRIN-1 antibody and immunostimulatory agent of the present invention is based on known methods such as mass spectrometry, ELISA, electrophoresis, and HPLC It can be characterized using
[0136]
　
　The conjugate of the anti -CAPRIN-1 antibody of the present invention and an immunostimulatory factor has cytotoxic activity in vitro or in vivo. Thus, the antitumor effect of the conjugate of the present invention can be known by examining the cytotoxic activity against cancer. The cytotoxic activity can be evaluated by administering the conjugate to a living organism having cancer, measuring the size of a tumor after administration, and examining the size of cancer over time. The antitumor effect of the present invention can also be evaluated by examining the survival rate. It can also be evaluated by examining the ability to produce cytokines or chemokines. The antitumor effect of the conjugate of the present invention can be further known by examining the prevention of cancer, the prevention of metastasis or the prevention of recurrence.
[0137]
　The conjugate of the present invention can be expected to have a stronger antitumor effect, as the binding affinity to the CAPRIN-1 protein on the surface of cancer cells is higher. The binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5 × 10 8 M -1 , at least 10 9 M -1 , at least 5 × 10 9 M −1 , at least 10 10 M −1 , at least 5 × 10 10 M −1 , at least 10 11 M −1 , at least 5 × 10 11 M −1 , at least 10 12 M -1 , or at least 10 13 M -1 is desirably.
[0138]
　The ability of the conjugate of the present invention to bind to CAPRIN-1 can be identified using, for example, a binding assay using ELISA, Western blotting, immunofluorescence and flow cytometry.
[0139]
　The conjugate of the present invention enhances the antitumor effect more than the anti-CAPRIN-1 antibody alone as described above, but the enhancement rate is preferably 30% or more, more preferably 40% or more, still more preferably 50% or more Still more preferably 55% or more, still more preferably 60% or more, even more preferably 65% ​​or more, and most preferably 70% or more. The enhancement ratio of the antitumor effect of the conjugate of the present invention to anti-CAPRIN-1 antibody alone is to compare the tumor volume on the 10th day and thereafter after administering the effective dose to tumor-bearing mice under the same conditions. It can be calculated by
[0140]
　 The target of the pharmaceutical composition for treating and / or preventing cancer of the present invention is particularly a cancer (cell) expressing a CAPRIN-1 protein It is not limited.
[0141]
　The terms "tumor" and "cancer" as used herein mean malignant neoplasm and are used interchangeably.
[0142]
　The cancer targeted by the present invention may be any cancer as long as it expresses CAPRIN-1 protein on the cell membrane surface. Preferably, the aforementioned breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cells Tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma. Testicular cancer, thyroid cancer, head and neck cancer.
[0143]
　More specifically, the cancer is, for example, breast cancer, complex type mammary adenocarcinoma, mammary gland mixed mixed tumor, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, nerve Epithelial tumors such as glioma, glioblastoma, neuroblastoma, endometriosis, neurogenic tumor, fetal neuroectodermal tumor, schwannoma, neurofibroma, meningioma, chronic Lymphocytic leukemia, Lymphoma, Gastrointestinal lymphoma, Gastrointestinal lymphoma, Small to medium cell lymphoma, Cecal cancer, Ascending colon cancer, Ascending colon cancer, Transverse colon cancer, Sigmoid colon cancer, Rectal cancer, Ovarian epithelium Cancer, germ cell tumor, stromal cell tumor, pancreatic ductal carcinoma, invasive pancreatic ductal carcinoma, adenocarcinoma of pancreatic carcinoma, acinar cell carcinoma, adenosquamous cell carcinoma, giant cell tumor, intraductal papillary mucinous neoplasm, mucinous cystic gland Cancer, Pancreatoblastoma, Pancreatocetocaroma, Frants tumor, Serous cyst adenocarcinoma, Solid papillary carcinoma, Gastrinoma, Glucagono Mammal, insulinoma, multiple endocrine neoplasia 1 (Wermer syndrome), non-functional islet cell tumor, somatostatoma, VIP producing tumor, cervical cancer, endometrial cancer, fibrosarcoma, bone and joint meat type, Ewing sarcoma, Wilms tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, soft tissue malignant tumor, teratoma group tumor, head and neck cancer, hypopharyngeal cancer, oropharyngeal cancer, tongue cancer, nasopharyngeal cancer, oral cavity It includes, but is not limited to, cancer, lip cancer, sinus cancer, laryngeal cancer and the like.
[0144]
　Preferred subjects (patients) to be targeted are mammals, for example, mammals including primates, pet animals, livestock animals, sport animals and the like, and humans, dogs and cats are particularly preferable.
[0145]
　When the conjugate used in the present invention is used as a pharmaceutical composition, it can be formulated by methods known to those skilled in the art. For example, it can be used parenterally in the form of an injection of a sterile solution or suspension with water or another pharmaceutically acceptable liquid. For example, a pharmacologically acceptable carrier or vehicle, specifically, sterile water or saline, vegetable oil, emulsifier, suspending agent, surfactant, stabilizer, fragrance, excipient, binder, etc. It is contemplated that it may be formulated by blending in unit dose form as required for generally accepted pharmaceutical practice, in appropriate combination with The amount of active ingredient in these preparations is such that a suitable dose in the indicated range can be obtained.
[0146]
　Sterile compositions for injection can be formulated according to a conventional formulation using a vehicle such as distilled water for injection. Aqueous solutions for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride, and suitable solubilizers, such as, for example, It may be used in combination with an alcohol, specifically ethanol, a polyalcohol such as propylene glycol, polyethylene glycol, a nonionic surfactant such as polysorbate 80 (TM), HCO-60. Examples of oily liquids include sesame oil and soybean oil, which may be used in combination with benzyl benzoate and benzyl alcohol as a solubilizing agent. It may also be formulated with a buffer such as phosphate buffer, sodium acetate buffer, a soothing agent such as procaine hydrochloride, a stabilizer such as benzyl alcohol, phenol, an antioxidant. The prepared injection is usually filled in a suitable ampoule. Examples of oily liquids include sesame oil and soybean oil, which may be used in combination with benzyl benzoate and benzyl alcohol as a solubilizing agent. It may also be formulated with a buffer such as phosphate buffer, sodium acetate buffer, a soothing agent such as procaine hydrochloride, a stabilizer such as benzyl alcohol, phenol, an antioxidant. The prepared injection is usually filled in a suitable ampoule.
[0147]
　The administration is oral or parenteral, preferably parenteral, and specifically includes injection, intranasal administration, pulmonary administration, transdermal administration and the like. Examples of the injection form can be administered systemically or locally, for example, by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, intratumoral injection and the like.
[0148]
　In addition, the administration method can be appropriately selected according to the age, weight, sex, symptoms and the like of the patient. The dose of the antibody or the pharmaceutical composition containing the antibody-encoding polynucleotide can be selected, for example, in the range of 0.0001 mg to 1000 mg per kg body weight. Alternatively, for example, the dose can be selected in the range of 0.001 to 100,000 mg / body per patient, but not necessarily limited to these values. The dose and administration method vary depending on the weight, age, sex, symptoms and the like of the patient, but can be appropriately selected by those skilled in the art.
[0149]
　A cancer, preferably a breast cancer or a renal cancer, which expresses the aforementioned CAPRIN-1 on the cell membrane surface by administering to a subject a pharmaceutical composition for treating and / or preventing a cancer comprising the conjugate of the present invention as an active ingredient. , Pancreatic cancer, colon cancer, lung cancer, brain cancer, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma. Testicular cancer, thyroid cancer, or head and neck cancer can be treated and / or prevented.
Example
[0150]
　EXAMPLES Hereinafter, the present invention will be specifically described based on examples, but the scope of the present invention is not limited by these specific examples.
[0151]
　Example 1 Anti-CAPRIN-1 Polyclonal Antibody
　Having Immunological Reactivity with CAPRIN-1 Protein Used for Anti-CAPRIN-1 Polyclonal Antibody Conjugate SEQ ID NO: 2 and SEQ ID NO: 4 prepared according to Example 3 of WO 2010/016526 1 mg of human CAPRIN-1 recombinant protein shown in (1) was mixed with an equal volume of incomplete Freund's adjuvant (IFA) solution, and this was subcutaneously administered to rabbits twice every two weeks. Blood was then collected to obtain antisera containing polyclonal antibodies. The obtained antiserum was purified using a protein G carrier (GE Healthcare Biosciences) to obtain a polyclonal antibody against CAPRIN-1 protein (anti-CAPRIN-1 polyclonal antibody # 1). In addition, sera from rabbits to which no antigen was administered were purified using a protein G carrier in the same manner as described above, and used as rabbit control antibodies.
[0152]
　Polyclonal antibodies # 2 to # 6 against the following partial polypeptides of CAPRIN-1 were obtained by the same method as the method for producing the above-mentioned polyclonal antibody against CAPRIN-1 protein.
[0153]
　SEQ ID NO: CAPRIN-1 protein The anti-CAPRIN-1 polyclonal antibody # 2 against the partial polypeptide represented by SEQ ID NO: 37 (SEQ ID NO: 31 in the present specification) described in WO 2011/096528, described in WO 2013/018894 Anti-CAPRIN-1 polyclonal antibody # 3 against the partial polypeptide represented by SEQ ID NO: 5 (SEQ ID NO: 32 in the present specification), SEQ ID NO: 5 described in WO2013 / 125654 (SEQ ID NO: 33 in the present specification) Anti-CAPRIN-1 polyclonal antibody # 4 against the partial polypeptide represented by SEQ ID NO: 37, anti-CAPRIN-1 polyclonal against the partial polypeptide represented by SEQ ID NO: 37 (SEQ ID NO: 34 in the present specification) Antibody # 5, WO2011 Anti caprin-1 polyclonal antibody against the partial polypeptide of SEQ ID NO: 37 listed (SEQ ID NO: 35 herein) to 096,534 # 6.
[0154]
　Example 2 Anti-CAPRIN-1 Monoclonal Antibody
　The following anti-CAPRIN-1 monoclonal antibody was used for the conjugate of the present invention.
[0155]
　A monoclonal antibody to CAPRIN-1 described in WO 2011/096528, 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 An antibody comprising CDRs 1 to 3 of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42 (for example, the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 39 containing CDR1 to 3 of the heavy chain variable region) An antibody comprising the amino acid sequence of the light chain variable region represented by SEQ ID NO: 43, which comprises CDRs 1 to 3 of the light chain variable region;
[0156]
　A monoclonal antibody to CAPRIN-1 described in WO2015 / 020212, 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 An amino acid sequence of a heavy chain variable region represented by SEQ ID NO: 47, comprising an antibody consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50 respectively (for example, CDR1 to 3 of the heavy chain variable region); An antibody comprising the amino acid sequence of the light chain variable region represented by SEQ ID NO: 51, which comprises the light chain variable regions CDR1 to 3;
[0157]
　A monoclonal antibody to CAPRIN-1 described in WO2011 / 096519, 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 An amino acid sequence of a heavy chain variable region represented by SEQ ID NO: 55 including the CDR1 to 3 of SEQ ID NO: 56, SEQ ID NO: 57 and SEQ ID NO: 58, for example, CDR1 to 3 of the heavy chain variable region; An antibody comprising the amino acid sequence of the light chain variable region represented by SEQ ID NO: 59, which comprises the light chain variable regions CDR1 to 3;
[0158]
　A monoclonal antibody to CAPRIN-1 described in WO2013 / 125654, 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 An amino acid sequence of a heavy chain variable region represented by SEQ ID NO: 63, comprising an antibody consisting of SEQ ID NO: 64, SEQ ID NO: 65 and SEQ ID NO: 66 respectively (for example, CDR1 to 3 of the heavy chain variable region) An antibody comprising the amino acid sequence of the light chain variable region represented by SEQ ID NO: 67, which comprises the light chain variable regions CDR1 to 3;
[0159]
　It is a monoclonal antibody against CAPRIN-1 described in WO2011 / 096517, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 68 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 69 An antibody comprising an amino acid sequence of a region.
[0160]
　A monoclonal antibody to CAPRIN-1 described in WO 2011/096528, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 70 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 71 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 72, and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 73; An antibody comprising an amino acid sequence of a heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 74 and an amino acid sequence of a light chain variable region represented by the amino acid sequence of SEQ ID NO: 75; Antibody comprising the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 77, An antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of issue 78, the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 79.
[0161]
　It is a monoclonal antibody against CAPRIN-1 described in WO 2011/096533, wherein the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 80 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 81 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 82 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 83.
[0162]
　A monoclonal antibody to CAPRIN-1 described in WO2011 / 096534, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 84 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 85 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 86 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 87.
[0163]
　A monoclonal antibody to CAPRIN-1 described in WO 2010/016526, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 88 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 89 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 90 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 91, An antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 92 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 93; Antibody comprising the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 95, Antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 96 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 97, represented by the amino acid sequence of SEQ ID NO: 98 An antibody comprising the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 99; the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 100; An antibody comprising the light chain variable region amino acid sequence represented by the amino acid sequence of SEQ ID NO: 101.
[0164]
　A monoclonal antibody to CAPRIN-1 described in WO 2013/018894, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 102 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 103 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 104 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 105.
[0165]
　A monoclonal antibody to CAPRIN-1 described in WO 2013/018892, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 106 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 107 An antibody comprising an amino acid sequence of a region.
[0166]
　A monoclonal antibody to CAPRIN-1 described in WO2013 / 018891, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 108 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 109 An antibody comprising an amino acid sequence of a region.
[0167]
　A monoclonal antibody to CAPRIN-1 described in WO2013 / 018889, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 111 An antibody comprising an amino acid sequence of a region.
[0168]
　A monoclonal antibody to CAPRIN-1 described in WO 2013/018883 which is an amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 112 and a light chain variable represented by the amino acid sequence of SEQ ID NO: 113 An antibody comprising an amino acid sequence of a region.
[0169]
　A monoclonal antibody to CAPRIN-1 described in WO 2013/125636, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 114 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 115 An antibody comprising an amino acid sequence of a region.
[0170]
　It is a monoclonal antibody against CAPRIN-1 described in WO2013 / 125654, and the light chain variable represented by the amino acid sequence of heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of SEQ ID NO: 117 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 118 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 119.
[0171]
　A monoclonal antibody to CAPRIN-1 described in WO 2013/125630, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 121 An antibody comprising an amino acid sequence of a region.
[0172]
　A monoclonal antibody against CAPRIN-1 described in WO2015 / 020212, which comprises the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 122 and the light chain variable represented by the amino acid sequence of SEQ ID NO: 123 An antibody comprising the amino acid sequence of the region, an antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 124, and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 125; An antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 127; Amino acid sequence of the heavy chain variable region, and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 129 Antibody comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 131; An antibody comprising the amino acid sequence of the heavy chain variable region represented by and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 133.
[0173]
　Among the above-mentioned anti-CAPRIN-1 monoclonal antibodies, CDRs 1 to 3 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 the framework region is a heavy chain comprising the sequence of human antibody The nucleotide sequence was designed so that the variable region could be expressed, and this was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 was inserted. Similarly, the nucleotide sequences are such that CDRs 1 to 3 of the light chain variable region are SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively, and the framework region can express the light chain variable region containing the sequence of human antibody. Was inserted into a mammalian expression vector into which the light chain constant region of human IgG1 was inserted. The above two recombinant expression vectors are introduced into mammalian cells according to a conventional method, and 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 A culture supernatant containing humanized monoclonal antibody # 1 (humanized antibody # 1) against CAPRIN-1 in which CDRs 1 to 3 consist of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively, was obtained.
[0174]
　Similarly, the heavy chain represented by SEQ ID NO: 47, wherein the 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 the framework region comprises the sequence of human antibody The nucleotide sequence was designed so that the variable region could be expressed, and this was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 was inserted. Similarly, the heavy chain represented by SEQ ID NO: 51, wherein the light chain variable region CDRs 1 to 3 consist of the amino acid sequences of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50 respectively, and the framework region comprises the sequence of human antibody The nucleotide sequence is designed so that the variable region can be expressed, and this is inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 has been inserted, and the above two recombinant expression vectors are introduced into mammalian cells according to a conventional method. Then, 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, and CDRs 1 to 3 of the light chain variable region respectively have SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50 A culture supernatant containing a humanized anti-CAPRIN-1 monoclonal antibody # 2 (humanized antibody # 2) comprising the amino acid sequence of
[0175]
　Similarly, CDR1-3 of the heavy chain variable region consist of the amino acid sequences of SEQ ID NO: 52, SEQ ID NO: 53 and SEQ ID NO: 54 respectively, and CDR1-3 of the light chain variable region respectively consist of SEQ ID NO: 56, SEQ ID NO: 57 and sequence A culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody # 3 (humanized antibody # 3) consisting of No. 58 was obtained.
[0176]
　Similarly, 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 respectively consist of SEQ ID NO: 64, SEQ ID NO: 65 and sequence A culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody # 4 (humanized antibody # 4) consisting of No. 66 was obtained.
[0177]
　Similarly, culture supernatants containing the following humanized anti-CAPRIN-1 monoclonal antibodies # 9 to # 41 (humanized antibodies # 9 to # 41) were obtained.
[0178]
　Humanized monoclonal antibody # 9 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 68 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 69 9).
[0179]
　Humanized monoclonal antibody # 10 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 70 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 71 10).
[0180]
　Humanized monoclonal antibody # 11 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 72 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 73 11).
[0181]
　Humanized monoclonal antibody # 12 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 75 12).
[0182]
　Humanized monoclonal antibody # 13 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 76 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 77 13).
[0183]
　Humanized monoclonal antibody # 14 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 78 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 79 14).
[0184]
　Humanized monoclonal antibody # 15 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 80 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 81 15).
[0185]
　Humanized monoclonal antibody # 16 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 82 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 83 16).
[0186]
　Humanized monoclonal antibody # 17 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 84 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 85 17).
[0187]
　Humanized monoclonal antibody # 18 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 86 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 87 18).
[0188]
　Antibody humanized monoclonal antibody # 19 (humanized antibody) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 88 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 89 # 19).
[0189]
　Humanized monoclonal antibody # 20 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 90 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 91 20).
[0190]
　Humanized monoclonal antibody # 21 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 92 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 93 21).
[0191]
　Humanized monoclonal antibody # 22 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 94 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 95 22).
[0192]
　Humanized monoclonal antibody # 23 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 96 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 97 23).
[0193]
　Humanized monoclonal antibody # 24 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 98 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 99 24).
[0194]
　Humanized monoclonal antibody # 25 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 100 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 101 25).
[0195]
　Humanized monoclonal antibody # 26 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 102 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 103 26).
[0196]
　Humanized monoclonal antibody # 27 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 104 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 105 27).
[0197]
　Humanized monoclonal antibody # 28 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 106 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 107 28).
[0198]
　Humanized monoclonal antibody # 29 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 108 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 109 29).
[0199]
　Humanized monoclonal antibody # 30 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 110 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 111 30).
[0200]
　Humanized monoclonal antibody # 31 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 112 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 113 31).
[0201]
　Humanized monoclonal antibody # 32 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 114 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 115 32).
[0202]
　Humanized monoclonal antibody # 33 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 116 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 117 33).
[0203]
　Humanized monoclonal antibody # 34 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 118 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 119 34).
[0204]
　Humanized monoclonal antibody # 35 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 120 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 121 35).
[0205]
　Humanized monoclonal antibody # 36 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 122 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 123 36).
[0206]
　Humanized monoclonal antibody # 37 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 124 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 125 37).
[0207]
　Humanized monoclonal antibody # 38 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 126 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 127 38).
[0208]
　Humanized monoclonal antibody # 39 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 128 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 129 39).
[0209]
　Humanized monoclonal antibody # 40 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 130 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 131 40).
[0210]
　Humanized monoclonal antibody # 41 (humanized antibody #) comprising the amino acid sequence of the heavy chain variable region represented by the amino acid sequence of SEQ ID NO: 132 and the amino acid sequence of the light chain variable region represented by the amino acid sequence of SEQ ID NO: 133 41).

The scope of the claims
[Claim 1]
　Immunological reactivity with an amino acid sequence represented by any one of SEQ ID NOs: 2 to 30 or an amino acid sequence having 80% or more sequence identity with the amino acid sequence And an antibody or fragment thereof, and a conjugate formed by binding an immunostimulatory factor.
[Claim 2]
　A CAPRIN- having the amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, 309, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. The conjugate according to claim 1, which has immunological reactivity with a partial polypeptide of 1 protein.
[Claim 3]
　The conjugate according to claim 1 or 2, wherein the antibody is a monoclonal antibody or a polyclonal antibody.
[Claim 4]

The complementation of 　conjugate (A) SEQ ID NO: 36, 37 and 38 according to any one of claims 1 to 3, wherein said antibody or fragment thereof is any of the following (A) to (M): A heavy chain variable region comprising the regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 40, 41 and 42 (CDR1, CDR2 and CD3 respectively) and
A heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 44, 45 and 46, respectively) and antibodies or fragments having immunological reactivity with 1 protein and SEQ ID NO: 48, 49 and 50 A light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of Antibody or fragment having immunological reactivity
complementarity (C) SEQ ID NO: 52, 53 and a complementary determining region 54 (respectively CDR1, CDR2 and CDR3) and the heavy chain variable region comprising SEQ ID NO: 56, 57 and 58
Complementarity of the antibody or fragment (D) SEQ ID NO: 60, 61 and 62, comprising a light chain variable region comprising the determining regions (CDR1, CDR2 and CDR3 respectively) and having immunological reactivity with CAPRIN-1 protein A heavy chain variable region comprising the determining region (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 64, 65 and 66 (CDR1, CDR2 and CDR3 respectively) and -1 antibody or fragment having immunological reactivity with protein
(E) The heavy chain variable region containing the complementarity determining regions of SEQ ID NOS: 170, 171 and 172 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NOS: 173, 174 and 175 (CDR1, CDR2 and CDR3 respectively) Antibody or an antibody having an immunological reactivity with the CAPRIN-1 protein, or a fragment thereof
(F) a complementarity-determining region of SEQ ID NO: 176, 177 and 178 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 179, 180 and 181 (CDR1, CDR2 and CDR3 respectively) and immunologically reactive with the CAPRIN-1 protein antibodies or fragments thereof having a
(G) SEQ ID NO: 182, 183 and 184 A heavy chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions of SEQ ID NO: 185, 186 and 187 (CDR1, CDR2 and CDR3 respectively), and
A heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 188, 189 and 190, respectively) of the antibody or fragment thereof (H) having immunological reactivity with CAPRIN-1 protein and SEQ ID NO: 191 , And a light chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of 192 and 193, and an antibody or fragment thereof having immunological reactivity with a CAPRIN-1 protein
(I) The heavy chain variable region containing the complementarity determining regions of SEQ ID NO: 146, 147 and 148 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 149, 150 and 151 (CDR1, CDR2 and CDR3 respectively) Antibody or an antibody having an immunological reactivity with the CAPRIN-1 protein or a fragment thereof
(J) the complementarity-determining regions of SEQ ID NO: 272, 273 and 274 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the complementarity determining regions of SEQ ID NO: 275, 276 and 277 (CDR1, CDR2 and CDR3 respectively) and immunologically reactive with CAPRIN-1 protein antibodies or fragments thereof having a
(K) SEQ ID NO: 290 and 291 and 292 A heavy chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) and a light chain variable region comprising complementarity determining regions of SEQ ID NO: 293, 294 and 295 (CDR1, CDR2 and CDR3 respectively), and
A heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3 of SEQ ID NO: 301, 302 and 303, respectively) of the antibody or fragment thereof (L) having immunological reactivity with CAPRIN-1 protein and SEQ ID NO: 305 , And a light chain variable region comprising complementarity determining regions (CDR1, CDR2 and CDR3 respectively) of 306 and 307, and an antibody or fragment thereof having immunological reactivity with a CAPRIN-1 protein
(M) The heavy chain variable region containing the complementarity determining regions of SEQ ID NO: 134, 135 and 136 (CDR1, CDR2 and CDR3 respectively) and the complementarity determining regions of SEQ ID NO: 137, 138 and 139 (CDR1, CDR2 and CDR3 respectively) And a light chain variable region comprising the antibody, and an antibody or fragment thereof having immunological reactivity with a CAPRIN-1 protein
[Claim 5]
　The conjugate according to any one of claims 1 to 4, wherein the antibody or fragment thereof is any of the following (a) to (ak):
(A) An antibody or fragment thereof wherein the
heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43 (b) the heavy chain variable region comprises SEQ ID NO: 47 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 51
(c) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 59 or a fragment thereof
(d) an antibody or 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 fragments thereof
comprise (e) the amino acid sequence of the heavy chain variable region SEQ ID NO: 68 and an antibody or its light chain variable region comprising the amino acid sequence of SEQ ID NO: 69 Fragment
(f) the heavy chain variable region comprises 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 antibody or fragment thereof
(g) the heavy chain variable region SEQ ID NO: 72 Wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73
(h) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 75
(I) 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 (j) The heavy chain variable region comprises SEQ ID NO: 78 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79
(k) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 81 or a fragment thereof
(1) An antibody or antibody 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 fragments thereof
comprising (m) is the amino acid sequence of the heavy chain variable region SEQ ID NO: 84 and an antibody or its light chain variable region comprising the amino acid sequence of SEQ ID NO: 85 Fragment
(n) a heavy chain variable region comprises 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 antibody or fragment thereof
(o) a heavy chain variable region SEQ ID NO: 88 Wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89, or the fragment
(p) of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90, and the light chain variable region An antibody or fragment thereof
(q) 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 Or a fragment thereof
(R) An antibody or fragment thereof in which the
heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 95 (s) An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 97
(t) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and the light chain variable region comprises An antibody comprising the amino acid sequence of SEQ ID NO: 99 or a fragment thereof
(u) an antibody or heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 101 fragments
anti which (v) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and light chain variable region comprising the amino acid sequence of SEQ ID NO: 103 Or fragments thereof
comprise (w) an amino acid sequence of the heavy chain variable region SEQ ID NO: 104, and antibodies or its fragments a light chain variable region comprising the amino acid sequence of SEQ ID NO: 105
(x) a heavy chain variable region sequence An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 106 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107
(y) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain An antibody or fragment thereof wherein the variable region comprises the amino acid sequence of SEQ ID NO: 109
(z) 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 Antibody or fragment thereof
(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 An antibody or fragment thereof comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115
(ac) The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116, and the light chain variable region comprises An antibody or fragment thereof
(ad) comprising the amino acid sequence of SEQ ID NO: 117, or an antibody or heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 118, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 119 fragments thereof
comprises an amino acid sequence of (ae) a heavy chain variable region SEQ ID NO: 120, and Ami light chain variable region SEQ ID NO: 121 Antibody or fragment thereof comprising an acid sequences
(af) a heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122, and antibodies or fragments thereof the light chain variable region comprising the amino acid sequence of SEQ ID NO: 123
(ag An antibody or a fragment thereof
(ah) 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; 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 or a fragment thereof
(ai) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128, and the light chain variable region comprises the SEQ ID NO: An antibody or fragment thereof comprising the amino acid sequence of 129
(Ai) An antibody or fragment thereof in which 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 (aj) An antibody or fragment thereof
(ak) comprising an amino acid sequence, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300, and An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 304.
[Claim 6]
　The conjugate according to any one of claims 1 to 5, wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single chain antibody.
[Claim 7]
　The immunostimulatory agent is a ligand or an agonist that binds to Toll-like receptor (TLR), NOD-like receptor (NLR), RIG-like receptor or C-type lectin receptor (CLR). The conjugate according to any one of the preceding claims.
[Claim 8]
　The conjugate according to claim 7, wherein the Toll-like receptor (TLR) is a ligand or agonist that binds to TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9 or TLR10.
[Claim 9]
　The conjugate according to claim 7 or 8, wherein the ligand or agonist that binds to the Toll-like receptor (TLR) is any one of the following (i) to (vii):
(I) a ligand or an agonist which binds to TLR2 selected from the group consisting of peptidoglycan, lipoprotein, lipopolysaccharide and zymosan
(ii) selected from the group consisting of Poly (I: C) and poly (A: U) A ligand or agonist that binds to TLR3
(iii) a ligand or agonist that binds to TLR4 selected from the group consisting of lipopolysaccharide (LPS), HSP60, RS09 and MPLA
(iv) is selected from the group consisting of flagellin and FLA Ligand or agonist that binds to
TLR5 (v) ligand or agonist that binds to TRL 7 or 8 selected from the group consisting of imidazoquinolines compounds and single-stranded RNA
(vi) bacterial DNA, unmethylated CpG DNA, hemozori a ligand or agonist that binds to TRL9 selected from the group consisting of n, ODN 1585, ODN 1668, ODN 1668 and ODN 1826
(vii) a ligand or agonist that binds to TRL 10 selected from the group consisting of profilin and uropathogenic bacteria
[Claim 10]
　The conjugate according to any one of claims 1 to 9, wherein the antibody or fragment thereof and the immunostimulatory agent are linked via a linker.
[Claim 11]
　A pharmaceutical composition for treating and / or preventing cancer, which comprises the conjugate according to any one of claims 1 to 10 as an active ingredient.
[Claim 12]
　The pharmaceutical composition according to claim 11, wherein the cancer is a cancer that expresses CAPRIN-1 protein on the cell membrane surface.
[Claim 13]
　The cancer includes breast cancer, kidney cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cells The pharmaceutical composition according to claim 11 or 12, which is tumor, melanoma, adrenocortical carcinoma, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, head and neck cancer.
[Claim 14]
　A method for treating and / or preventing cancer, which comprises administering to a subject the conjugate according to any one of claims 1 to 10, the pharmaceutical composition according to any one of claims 11 to 13 .