Title of Invention: Pharmaceutical Composition for Treatment and / or Prevention of Cancer
Technical field
[0001]
　The present invention relates to an antibody against the CAPRIN-1 protein or a fragment thereof and a drug for treating and / or preventing cancer using imiquimod.
Background technology
[0002]
　Various antibody drugs targeting specific antigen proteins on cancer cells have been applied to cancer treatment as cancer therapeutic agents having few side effects due to their cancer specificity. For example, on the cell membrane surface of many solid cancers, Cytopasmic-activation and proliferation-associated patent 1 (CAPRIN-1) is expressed, and an antibody against this CAPRIN-1 protein is used for cancer treatment and / or preventive medicine. It is known to be promising (Patent Document 1).
[0003]
　In recent years, in clinical practice, a treatment method in which a plurality of cancer therapeutic agents are used in combination has been used as a standard therapeutic method in order to enhance the effectiveness of the cancer therapeutic agent. For example, for colon cancer, a combination of irinotecan, phoric acid, and fluorouracil, for breast cancer, a combination of doxorubicin and cyclophosphamide, or for paclitaxel, trastuzumab, bertustumab, and gastric cancer. Is commonly treated with multiple anti-cancer agents such as cisplatin and fluorouracil. It has been confirmed that a cancer therapeutic agent containing an anti-CAPRIN-1 antibody as an active ingredient also has an excellent cancer therapeutic effect when used in combination with a chemotherapeutic agent (Patent Document 2). However, the treatment of cancer by a combination of chemotherapeutic agents is not effective for all applicable cancers, and although the therapeutic effect may be additively enhanced, it is synergistically significant. There are few things that enhance the therapeutic effect.
[0004]
　Imiquimod is known as an agonist of Toll-like receptors (TLRs) 7 or 8. The main mechanism of action of imikimod in vivo is to suppress virus growth through promotion of IFN-α production and to damage virus-infected cells by activating cellular immune response, thereby treating diseases associated with virus infection. It is considered to be effective, and based on this mechanism of action, it is a topical drug that was originally approved as a therapeutic drug for condyloma acuminata rather than a therapeutic drug for cancer. Since then, its effectiveness has been confirmed for superficial diseases such as actinic keratosis and Bowen's disease. In Europe and the United States, imiquimod is also approved for superficial basal cell carcinoma, and is applied for the treatment of superficial skin cancer (Boen's disease, melanoma, cutaneous T cell lymphoma) (non-patented). Document 1). One of the mechanisms of action on superficial basal cell carcinoma is thought to be the strong activation of the innate immune system by monocytes and macrophages, which are one of the immune cells in the body (non-). Patent Document 2). However, imiquimod has only been approved for use in some cancers (superficial basal cell carcinoma). In addition, imiquimod is known to be used as a symptomatic treatment for extramammary paget disease, and Cohen et al. Reported that 7 out of 9 cases were completely successful (Non-Patent Document 3).
[0005]
　In addition, a method for treating cancer by combining a cancer antibody drug and a factor that activates an antigen-presenting cell containing an agonist of a Toll-like receptor (Patent Document 3), and a targeted therapeutic agent containing an antibody drug (targeted therapy). ) And an immunotherapeutic agent (immunotherapeutic) capable of activating human trait cell-like dendritic cells, myeloid dendritic cells, or NK cells in combination (combination) to treat cancer (Patent Document 4). Imikimod is described as an example of the above-mentioned factors that activate antigen-presenting cells and immunotherapeutic agents. As described above, although there are documents suggesting a cancer treatment method by using a combination of a cancer antibody drug and an imiquimod (combination), these documents include data on the cancer treatment effect when the combination of an imiquimod and a cancer antibody drug (combination) is actually performed. Has not been shown and its effectiveness has not been clarified. Rather, for basal cell carcinoma that occurred in patients with a history of administration of rituximab (a monoclonal antibody that specifically binds to the CD20 protein expressed on the surface of cancer cells), which is a type of cancer antibody drug for the treatment of non-Hodgkin's lymphoma. On the other hand, it has been reported that when imikimod is administered, the antitumor effect of imikimod is attenuated (Non-Patent Document 4).
Prior art literature
Patent documents
[0006]
Patent Document 1: WO2010 / 016526
Patent Document 2: WO2011 / 0965535
Patent Document 3: WO2015 / 112749
Patent Document 4: WO2016 / 00475
Non-patent literature
[0007]
Non-Patent Document 1: Skin Therapy Lett. 2007, 7, 1-6
Non-Patent Document 2: British J Dermatol. 2003, 149, 57-58
Non-Patent Document 3: South Med J, 2006,99,396-402
Non-Patent Document 4: Journal of Medical Case Reports (2016) 10:57
Outline of the invention
Problems to be solved by the invention
[0008]
　An object of the present invention is to provide a pharmaceutical composition for treating and / or preventing a cancer that specifically expresses the CAPRIN-1 protein on the cell surface.
Means to solve problems
[0009]
　As mentioned above, basal cells generated in patients with a history of administration of rituximab (a monoclonal antibody that specifically binds to the CD20 protein expressed on the surface of cancer cells), which is a type of cancer antibody drug for the treatment of non-hodgkinous lymphoma. It has been reported that when imikimod is administered to cancer, the antitumor effect of imikimod is diminished. However, in this report, some cancer antibody drugs targeting cancer cells are used in combination with imikimod. It suggested to those skilled in the art that the antitumor effect of the disease might be diminished. However, as a result of diligent research, the present inventor has determined that an antibody against the CAPRIN-1 protein, which has immunological reactivity with cancer cells, or a fragment thereof and imikimod are used in combination (combination) with the antibody against the CAPRIN-1 protein or a fragment thereof alone. Alternatively, it exerts an extremely strong antitumor effect as compared with imikimod alone, and further, the effect of enhancing the antitumor effect when an antibody against the CAPRIN-1 protein or a fragment thereof is used in combination with imikimod is an existing cancer antibody drug and imikimod. Is significantly superior to the effect of enhancing the antitumor effect when used in combination with, and in addition, the effect of enhancing the antitumor effect when an antibody against the CAPRIN-1 protein or a fragment thereof is used in combination with imikimod is different from that of imikimod. We have found that it is superior to the antitumor effect when an existing anticancer agent is used in combination, and have completed the present invention.
[0010]
　Specifically, the present invention has the following features (1) to (14).
[0011]
　(1) A drug for treating and / or preventing cancer, which comprises an antibody or fragment thereof having immunological reactivity with the CAPRIN-1 protein and imiquimod in combination or separately. ..
[0012]
　(2) CAPRIN in which the antibody or fragment thereof has an amino acid sequence represented by any of SEQ ID NOs: 2 to 30, which is an even number, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. -1 The drug according to (1), which has immunological reactivity with a protein.
[0013]
　(3) The drug according to (1) or (2), wherein the antibody or fragment thereof has immunological reactivity with the extracellular region of CAPRIN-1 protein present on the surface of cancer cells.
[0014]
　(4) 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, and 309, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. The pharmaceuticals according to (1) to (3), which have immunological reactivity with a partial polypeptide of the CAPRIN-1 protein having.
[0015]
　(5) The drug according to any one of (1) to (4), wherein the antibody is a monoclonal antibody or a polyclonal antibody.
[0016]
　(6) The drug according to any one of (1) to (5), wherein the antibody or fragment thereof is any of the following (A) to (M).
(A) Heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 36, 37 and 38 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 40, 41 and 42 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments (B) SEQ ID NOs: 44, 45 and 46 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable regions and are immunologically reactive with the CAPRIN-1 protein. Contains heavy chain variable regions containing, and light chain variable regions containing complementarity determining regions of SEQ ID NOs: 48, 49 and 50 (CDR1, CDR2 and CDR3, respectively), and immunoreactivity with CAPRIN-1 proteins. The
heavy chain variable regions containing the complementarity determining regions of antibody or fragment (C) SEQ ID NOs: 52, 53 and 54 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 56, 57 and 58 (CDR1, respectively).
The complementarity determining regions of antibodies or fragments (D) SEQ ID NOs: 60, 61 and 62 (CDR1, respectively, ) comprising a light chain variable region containing CDR2 and CDR3) and immunoreactive with the CAPRIN-1 protein . It contains heavy chain variable regions containing CDR2 and CDR3) and light chain variable regions containing complementarity determining regions of SEQ ID NOs: 64, 65 and 66 (CDR1, CDR2 and CDR3, respectively), and CAPRIN-1 protein and immunology. Reactive antibodies or fragments
(E) Heavy chain variable regions containing complementarity determining regions 170, 171 and 172 (CDR1, CDR2 and CDR3 respectively) and complementarity determining regions of SEQ ID NOs: 173, 174 and 175 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments thereof (F) SEQ ID NOs: 176, 177 and 178 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable region and have immunoreactivity with the CAPRIN-1 protein. ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 (CDR1, CDR2 and CDR3, respectively) and immunoreactivity with the CAPRIN-1 protein. Antibodies or fragments thereof
(G) with heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 182, 183 and 184 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 185, 186 and 187, respectively.
The complementarity determining regions (H) SEQ ID NOs: 188, 189 and 190 of an antibody or fragment (H) thereof that contains light chain variable regions containing CDR1, CDR2 and CDR3) and is immunologically reactive with the CAPRIN-1 protein. The heavy chain variable regions containing CDR1, CDR2 and CDR3) and the light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively), and the CAPRIN-1 protein. Antibodies or fragments thereof that are immunologically reactive with
(I) Heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 146, 147 and 148 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 149, 150 and 151 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments thereof (J) SEQ ID NOs: 272, 273 and 274 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable region containing the CAPRIN-1 protein and have immunoreactivity with the CAPRIN-1 protein. ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 275, 276 and 277 (CDR1, CDR2 and CDR3, respectively) and immunoreactivity with the CAPRIN-1 protein. Antibodies or fragments thereof
(K) with heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 290, 291 and 292 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 293, 294 and 295, respectively.
The complementarity determining regions (L) of the antibody or fragment (L) SEQ ID NOs: 301, 302 and 303 containing a light chain variable region containing CDR1, CDR2 and CDR3) and immunoreactive with the CAPRIN-1 protein. The heavy chain variable regions containing CDR1, CDR2 and CDR3) and the light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 305, 306 and 307 (CDR1, CDR2 and CDR3, respectively), and the CAPRIN-1 protein. Antibodies or fragments thereof that are immunologically reactive with
(M) Heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 134, 135 and 136 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 137, 138 and 139 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region comprising, and has immunological reactivity with the CAPRIN-1 protein.
[0017]
　(7) The drug according to any one of (1) to (6), wherein the antibody or fragment thereof is any of the following (a) to (ak).
(A) An antibody or a fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 39 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 43. (B) The heavy chain variable region of SEQ ID NO: 47. An antibody or fragment thereof comprising an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 51
(c) The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 55 and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 59
(d) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 63 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 67. An
antibody in which the fragment (e) heavy chain variable region contains the amino acid sequence of SEQ ID NO: 68 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 69 or a fragment
(f) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(g) containing 70 amino acid sequences and the light chain variable region containing the amino acid sequence of SEQ ID NO: 71. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 72 and the light chain variable. An antibody or fragment thereof
(h) in which the region comprises the amino acid sequence of SEQ ID NO: 73, 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. Antibody or fragment thereof
(I) An antibody or fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 76 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 77. (J) The heavy chain variable region of SEQ ID NO: 78. An antibody or fragment thereof
(k) containing an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 79. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 80, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 81
(l) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 82 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 83. An
antibody in which the fragment (m) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 85 or a fragment
(n) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(o) containing the amino acid sequence of 86 and the light chain variable region containing the amino acid sequence of SEQ ID NO: 87. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 88 and the light chain variable. An antibody or fragment thereof
(p) in which the region comprises the amino acid sequence of SEQ ID NO: 89 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. Antibody or fragment thereof
(q) An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 92 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 93.
(R) An antibody or fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 94 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 95. (S) The heavy chain variable region of SEQ ID NO: 96. An antibody or fragment thereof
(t) containing an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 97. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 98, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 99.
(U) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 100 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 101. The fragment
(v) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 103, or the fragment
(w) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(x) containing 104 amino acid sequences and the light chain variable region containing the amino acid sequence of SEQ ID NO: 105 contains the amino acid sequence of SEQ ID NO: 106 and the light chain variable region An antibody or fragment thereof
(y) in which the region comprises the amino acid sequence of SEQ ID NO: 107. 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. Antibody or fragment thereof
(z) An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 110 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 111.
(Aa) An antibody or fragment thereof
(ab) in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 113 (ab) The heavy chain variable region of SEQ ID NO: 114. An antibody or fragment
(ac) thereof comprising an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 115 The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 116, and the light chain variable region An antibody comprising the amino acid sequence of SEQ ID NO: 117 or a fragment thereof
(ad) An antibody or an antibody in which 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. An
antibody in which the fragment (ae) heavy chain variable region contains the amino acid sequence of SEQ ID NO: 120 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 121 or a fragment
(af) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(ag) containing the amino acid sequence of 122 and having the light chain variable region containing the amino acid sequence of SEQ ID NO: 123 The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 124 and the light chain variable The antibody or fragment
(ah) heavy chain variable region comprising the region comprising the amino acid sequence of SEQ ID NO: 125 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. Antibody or fragment thereof
(ai) An antibody or fragment thereof comprising a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 128 and a light chain variable region containing the amino acid sequence of SEQ ID NO: 129.
(Ai) An antibody in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 131 or a fragment thereof
(aj) The heavy chain variable region of SEQ ID NO: 132 An antibody comprising an amino acid sequence and the light chain variable region contains the amino acid sequence of SEQ ID NO: 133 or a fragment
(ak) thereof The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 300 and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 304.
[0018]
　(8) The drug according to any one of (1) to (7), wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single chain antibody.
[0019]
　(9) The drug according to any one of (1) to (8), wherein the cancer is a cancer that expresses the CAPRIN-1 protein on the cell membrane surface.
[0020]
　(10) The cancers are basal cell cancer, budget disease, skin cancer, breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia. , Lymphoma, liver cancer, bile sac cancer, sarcoma, obesity cell tumor, corticolytic cancer, Ewing tumor, Hodgkin lymphoma, mesopharyngeal tumor, multiple myeloma, testicle cancer, thyroid cancer or head and neck cancer, (1)- The drug according to any one of (9).
[0021]
　(11) The drug according to any one of (1) to (10), wherein the dosage form of the imiquimod is a transdermal administration type preparation.
[0022]
　(12) An agent for enhancing the efficacy of a pharmaceutical composition for treating and / or preventing cancer, which comprises an antibody or a fragment thereof having immunological reactivity with the CAPRIN-1 protein as an active ingredient and containing imiquimod as an active ingredient.
[0023]
　(13) An agent for enhancing the efficacy of a pharmaceutical composition for treating and / or preventing cancer containing imiquimod as an active ingredient, which comprises an antibody having immunological reactivity with the CAPRIN-1 protein or a fragment thereof as an active ingredient.
[0024]
　(14) For the treatment and / or prevention of cancer, which comprises administering an antibody or a fragment thereof having immunological reactivity to the CAPRIN-1 protein and imiquimod to a subject together or separately. the method of.
The invention's effect
[0025]
　The combined use of an antibody against the CAPRI-1 protein or a fragment thereof according to the present invention and imiquimod not only exerts a stronger antitumor effect than the antibody against the CAPRIN-1 protein alone and imiquimod alone, but also an antibody drug against existing cancers. Shows superior antitumor effect than the combined use of imiquimod and imiquimod. Furthermore, the combined use of the antibody against the CAPRI-1 protein according to the present invention and imiquimod shows a stronger antitumor effect as compared with the combined use of the existing chemotherapeutic agent and the antibody against the CAPRIN-1 protein. Therefore, the combined use of an antibody against the CAPRIN-1 protein and imiquimod is effective in the treatment and prevention of cancer.
Mode for carrying out the invention
[0026]
　The antitumor activity of an antibody against the CAPRIN-1 protein used in the present invention or a fragment thereof (hereinafter referred to as "anti-CAPRIN-1 antibody") in combination with imikimod is a tumor for a cancer-bearing animal in vivo as described later. It can be evaluated by examining the inhibition of proliferation.
[0027]
　As used herein, the term "combination" or "combination" refers to the administration of an anti-CAPRIN-1 antibody and imiquimod to the same living body simultaneously or at predetermined intervals as independent active ingredients. The interval may be simultaneous administration, after 30 minutes, or after 1 hour, 3 hours, 6 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, 2 weeks, 3 It may be after a week or four weeks. When the anti-CAPRIN-1 antibody or imiquimod exhibits its activity in vivo, either one may be administered. Further, the anti-CAPRIN-1 antibody may be administered first, or imiquimod may be administered first.
[0028]
　The anti-CAPRIN-1 antibody according to the present invention may be a monoclonal antibody or a polyclonal antibody, preferably a monoclonal antibody, and is any kind of antibody as long as the antibody of the present invention can exhibit antitumor activity. The antibody may be a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, or a non-human animal antibody.
[0029]
　Further, the subject to be treated and / or prevented from cancer in the present invention is a mammal such as a human, a pet animal, a domestic animal, or a competition animal, and a preferable subject is a human.
[0030]
　The anti-CAPRIN-1 antibody, imiquimod, a drug containing them as an active ingredient, and a method for treating and / or preventing cancer according to the present invention will be described below.
[0031]
　
　It is represented by any of an even number of SEQ ID NOs: 2 to 30, which is a specific example of an amino acid having immunological reactivity with the anti-CAPRIN-1 antibody used in the present invention. Among the CAPRIN-1 proteins having an amino acid sequence, the amino acid sequences shown by SEQ ID NOs: 6, 8, 10, 12 and 14 are the amino acid sequences of the canine CAPRIN-1 protein, and the amino acid sequences shown by SEQ ID NOs: 2 and 4. Is the amino acid sequence of human CAPRIN-1 protein, the amino acid sequence shown by SEQ ID NO: 16 is the amino acid sequence of bovine CAPRIN-1 protein, and the amino acid sequence shown by SEQ ID NO: 18 is that of horse CAPRIN-1 protein. It is an amino acid sequence, and the amino acid sequence shown by SEQ ID NOs: 20 to 28 is the amino acid sequence of mouse CAPRIN-1 protein, and the amino acid sequence shown by SEQ ID NO: 30 is the amino acid sequence of chicken CAPRIN-1 protein.
[0032]
　The anti-CAPRIN-1 antibody used in the present invention is 80% or more, preferably 90% or more, more preferably 95% or more, preferably 90% or more, more preferably 95% or more of the amino acid sequence represented by any of the even-numbered SEQ ID NOs: 2 to 30. It may be immunologically reactive with a variant of the CAPRIN-1 protein having a sequence identity of% or more, more preferably 99% or more. The term "% sequence identity" as used herein refers to the amino acid (or base) when two sequences are aligned so as to have the maximum similarity with or without a gap. It means the percentage (%) of the same amino acid (or base) to the total number.
[0033]
　In the present invention, the anti-CAPRIN-1 antibody means an antibody or a fragment thereof having immunological reactivity with the full length of the CAPRIN-1 protein or a fragment thereof. Here, "immunological reactivity" means a property that an antibody binds to a CAPRIN-1 protein or a partial polypeptide thereof in vivo.
[0034]
　The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.
[0035]
　A polyclonal antibody (anti-CAPRIN-1 polyclonal antibody) having immunological reactivity with the full length of the CAPRIN-1 protein or a fragment thereof is, for example, a natural CAPRIN-1 protein, a fusion protein with GST, or a partial peptide thereof. Immunize mice, human antibody-producing mice, rats, rabbits, chickens, etc. to obtain serum. The obtained serum can be obtained by sulfite precipitation, protein A, protein G, DEAE ion exchange column, affinity column to which CAPRIN-1 protein or partial peptide is bound, or the like.
[0036]
　For the full length of the CAPRIN-1 protein used for immunization or a fragment thereof, the nucleotide sequence and amino acid sequence of CAPRIN-1 and its homologue can be obtained by accessing, for example, GenBank (NCBI, USA) and using algorithms such as BLAST and FASTA (Karlin and Altschul). , Proc. Natl. Acad. Sci. USA, 90: 5873-5877, 1993; Altschul et al., Nucleic Acids Res. 25: 3389-3402, 1997). Further, the method for producing the CAPRIN-1 protein can be obtained by referring to WO2014 / 012479, and cells expressing the CAPRIIN-1 protein or the like can also be used.
[0037]
　Monoclonal antibodies (anti-CAPRIN-1 monoclonal antibodies) that have immunological reactivity with the full length of the CAPRIN-1 protein or fragments thereof are, for example, those of the breast cancer cells SK-BR-3 and CAPRIN-1 proteins that express CAPRIN-1. A clone that produces an anti-CAPRIN-1 monoclonal antibody is selected from the obtained fusion cells (hybridoma) by fusing the spleen cells and myeloma cells isolated from the mice by administering the full length or a fragment thereof to the mice to immunize them. Can be obtained by The antibody produced from the selected hybridoma can be obtained by the same method as the above-mentioned method for purifying the polyclonal antibody.
[0038]
　Antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies, and non-human animal antibodies.
[0039]
　The human antibody sensitizes human lymphocytes infected with EB virus with proteins, protein-expressing cells or lysates thereof, and fuses the sensitized lymphocytes with myeloma cells such as human-derived U266 cells, resulting in fusion. Antibodies having immunoreactivity with the full length of the CAPRIN-1 protein or fragments thereof can be obtained from cells.
[0040]
　A humanized antibody is a modified antibody, also referred to as a restored human antibody. Humanized antibodies are constructed by transplanting the complementarity determining regions of an immune animal-derived antibody into the complementarity determining regions of a human antibody. 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 regions of mouse antibody and rabbit antibody and the framework regions of human antibody are prepared so as to have an overlapping portion at the terminal portion. Synthesize from the oligonucleotides of the above by the PCR method. It is obtained by ligating the obtained DNA with a DNA encoding a constant region of a human antibody, incorporating it into an expression vector, and introducing it into a host for production (European Patent Application Publication No. EP239400, International Publication No. See WO 96/02576). The framework regions of human antibodies linked via the complementarity determining regions are selected so that the complementarity determining regions form a good antigen binding site. If desired, amino acids in the framework regions in the variable region of the antibody may be substituted so that the complementarity determining regions of the reconstituted human antibody form the appropriate antigen binding site (Sato K. et al., Cancer Research). 1993, 53: 851-856). It may also be replaced with framework regions derived from various human antibodies (see WO 99/51743).
[0041]
　Antibodies are heteropolyglycoproteins, usually containing at least two heavy chains and two light chains. The antibody is composed of two identical light chains and two identical heavy chains. The heavy chain has a heavy chain variable region at one end, followed by several constant regions. The light chain has a light chain variable region at one end, followed by several constant regions. The variable regions exhibit specific variable regions called complementarity determining regions (CDRs) to impart binding specificity to the antibody. The portion that is relatively conserved in the variable region is called the framework region (FR). The variable regions of the complete heavy and light chains each contain four FRs linked by three CDRs (CDR1 to CDR3).
[0042]
　The sequences of the constant region and variable region of human-derived heavy and light chains can be obtained from, for example, NCBI (USA: GenBank, UniGene, etc.). For example, the heavy chain constant region of human IgG1 is registered in Registration No. J00228. For the heavy chain constant region of human IgG2, refer to the sequence of registration number J00230, for the human light chain κ constant region, reference to the sequences of registration numbers V00557, X64135, X64133, etc., and for the human light chain λ constant region, refer to the sequences of registration numbers X64132, X64134, etc. be able to.
[0043]
　A chimeric antibody is an antibody produced by combining sequences derived from different animals, for example, from the constant regions of the heavy chain variable region and the light chain variable region of a mouse antibody and the heavy chain variable region and the light chain variable region of a human antibody. Antibodies and the like. The chimeric antibody can be prepared by 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 ligated, and this is incorporated into an expression vector and introduced into a host. Obtained by producing.
[0044]
　Non-human animal antibodies are obtained by injecting the sensitizing antigen intraperitoneally, intradermally or subcutaneously in animals such as mice as a general method of immunizing an animal with a sensitizing antigen according to a known method. .. When injecting a sensitizing antigen, an appropriate amount is mixed with various adjuvants such as CFA (Freund Complete Adjuvant) and administered to animals multiple times. After immunizing the animal and confirming that the serum contained the anti-CAPRIN-1 antibody, the serum was obtained, and as described above, sulfate precipitation, protein A, protein G, DEAE ion exchange column, CAPRIN-1 protein. It can be obtained by an affinity column or the like to which a partial peptide is bound. In addition, when a monoclonal antibody is obtained from a non-human animal, it can be obtained by collecting immune cells from an immunized animal and subjecting them to cell fusion with myeloma cells. Cell fusion between the immune cells and myeloma cells can be performed according to a known method (see Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).
[0045]
　The antibody used in the present invention can also be obtained as a recombinant antibody produced by cloning an antibody gene from a hybridoma, incorporating it into an appropriate vector, introducing it into a host, and producing it using a gene recombination technique. Can (Carl, AK Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISH).
[0046]
　The anti-CAPRIN-1 antibody used in the present invention may be one in which an amino acid in a variable region (for example, FR) or a constant region is replaced with another amino acid. The amino acid substitution is one or more, for example, less than 15, less than 10, 8 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less amino acids, preferably 1 to 9 amino acids. The antibody should be an antibody that has the property of specifically binding to an antigen, the binding affinity to an antigen is equal to or higher than that of an unsubstituted antibody, and does not cause rejection when applied to humans. .. Amino acid substitutions are preferably conservative amino acid substitutions, which are substitutions between amino acids with similar properties such as charge, side chain, polarity, and aromaticity. Amino acids with similar properties include, for example, basic amino acids (arginine, lysine, histidine), acidic amino acids (aspartic acid, glutamic acid), uncharged polar amino acids (glycine, asparagine, glutamine, serine, threonine, cysteine, tyrosine), non-polar amino acids. It can be classified into sex amino acids (leucine, isoleucine, alanine, valine, proline, phenylalanine, tryptophan, methionine), branched amino acids (threonine, valine, isoleucine), aromatic amino acids (phenylalanine, tyrosine, tryptophan, histidine) and the like.
[0047]
　The anti-CAPRIN-1 antibody used in the present invention can be expected to have a stronger antitumor effect when it has a high binding affinity with the CAPRIN-1 protein on the surface of cancer cells. 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 , and at least 5 ×. 10 9 M -1 , at least 10 10 M -1 , at least 5 x 10 10 M -1 , at least 10 11 M -1 , at least 5 x 10 11 M -1 , at least 10 12It is preferably M -1 , or at least 10 13 M -1 .
[0048]
　The anti-CAPLIN-1 antibody used in the present invention may be chemically modified, and examples of such antibody modifications include polyethylene glycol (PEG) and antitumor compounds (eg, antitumor compounds exemplified below). Antibodies bound to various molecules such as tumor agents) can be mentioned. In the antibody modified product of the present invention, the substance to be bound is not limited. Such an antibody-modified product can be obtained by chemically modifying the obtained antibody. These methods have already been established in this field. The anti-CAPRIN-1 antibody used in the present invention replaces one, two or several amino acids in the heavy chain constant region of the antibody, or N- in an N-glycosidic bond sugar chain that binds to the heavy chain constant region. By removing the fucose bound to acetylglucosamine, the binding force of the anti-CAPRIN-1 antibody to the effector cells can be improved. The above may be an amino acid substitution alone, or may be a composition with an antibody to which fucose is bound.
[0049]
　Antibodies in which one, two, or several amino acids in the heavy chain constant region are substituted are prepared by referring to, for example, WO2004 / 063351, WO2011 / 120135, US Pat. No. 8,388,955, WO2011 / 005481, US Pat. No. 6,737,056, WO2005 / 063351. can do.
[0050]
　For an antibody or cells in which fucose attached to N-acetylglucosamine in an N-glycosidic bond sugar chain in a heavy chain constant region has been removed, refer to US Pat. No. 6,602,684, European Patent No. 1914244, and US Pat. No. 7,579,170. Can be produced. The composition of the antibody from which fucose bound to N-acetylglucosamine in the N-glycosidic bond sugar chain bound to the heavy chain constant region and the antibody to which fucose is bound, or the producing cells thereof, are described in, for example, US Pat. It can be made by referring to the number.
[0051]
　The anti-CAPRIN-1 polyclonal antibody, anti-CAPRIN-1 monoclonal antibody used in the present invention, the method for producing the antibody, the purification method, and the method for producing the CAPRIN-1 protein or a partial polypeptide thereof used for immunization are described in WO2010 / 016526 and WO2011 /. 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 It can be obtained with reference to WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212.
[0052]
　Specific examples of the anti-CAPRIN-1 antibody in the present invention include the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 /. 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212. -1 antibody is mentioned, and the following are mentioned as preferable anti-CAPRIN-1 antibody.
[0053]
　80% or more of the amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4 or the amino acid sequence (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, still more preferably 99% or more). An antibody or fragment thereof that is immunologically reactive with a partial polypeptide of the CAPRIN-1 protein having an amino acid sequence having the sequence identity of.
[0054]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 31 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 36, 37 and 38 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 40, 41 and 42 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that contains a light chain variable region and is immunologically reactive with the CAPRIN-1 protein, or complementarity determining regions of SEQ ID NOs: 140, 141 and 142 (CDR1, CDR2 and CDR3, respectively). Contains heavy chain variable regions containing, and light chain variable regions containing complementarity determining regions of SEQ ID NOs: 143, 144 and 145 (CDR1, CDR2 and CDR3, respectively), and immunologically reactive with the CAPRIN-1 protein. An antibody or fragment thereof, or a heavy chain variable region containing the complementarity determining regions of SEQ ID NOs: 164, 165 and 166 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 167, 168 and 169 (CDR1 respectively). , CDR2 and CDR3), and an antibody or fragment thereof that comprises the light chain variable regions and has immunological reactivity with the CAPRIN-1 proteins. More preferably, 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 a fragment thereof, or the heavy chain variable region is SEQ ID NO: 70. An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 71 and the light chain variable region containing the amino acid sequence of SEQ ID NO: 71, or a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 78 and a light chain variable region. Is an antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 79.
[0055]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 33 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 60, 61 and 62 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 64, 65 and 66 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region comprising, and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 67.
[0056]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 32 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 52, 53 and 54 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 56, 57 and 58 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region comprising, and has immunological reactivity with the 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.
[0057]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 34 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions 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). An antibody or fragment thereof that contains a light chain variable region containing, and is immunologically reactive with the CAPRIN-1 protein, or complementarity determining regions of SEQ ID NOs: 176, 177, and 178 (CDR1, CDR2, and CDR3, respectively). ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 (CDR1, CDR2 and CDR3, respectively) and immunologically reactive with the CAPRIN-1 protein. Antibodies or fragments thereof. More preferably, the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 80 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 81, or a fragment thereof, or the heavy chain variable region is SEQ ID NO: 82. An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 83 and the light chain variable region containing the amino acid sequence of SEQ ID NO: 83.
[0058]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 35 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 182, 183 and 184 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 185, 186 and 187 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that contains a light chain variable region containing, and is immunologically reactive with the CAPRIN-1 protein, or complementarity determining regions of SEQ ID NOs: 188, 189 and 190 (CDR1, CDR2 and CDR3, respectively). ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively) and immunologically reactive with the CAPRIN-1 protein. Antibodies or fragments thereof. More preferably, an 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 contains the amino acid sequence of SEQ ID NO: 85, or the heavy chain variable region is SEQ ID NO: 86. An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 87 and in which the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87.
[0059]
　Heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 44, 45 and 46 (CDR1, CDR2 and CDR3, respectively) and light including complementarity determining regions of SEQ ID NOs: 48, 49 and 50 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a chain variable region and is immunologically reactive with the 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.
[0060]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 296 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 146, 147 and 148 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 149, 150 and 151 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the 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.
[0061]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 297 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 272, 273 and 274 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 275, 276 and 277 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises light chain variable regions and has immunological reactivity with the 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.
[0062]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 298 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 290, 291 and 292 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 293, 294 and 295 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121.
[0063]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 299 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 301, 302 and 303 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 305, 306 and 307 (CDR1, CDR2 and CDR3, respectively) An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the 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.
[0064]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 308 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 134, 135 and 136 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 137, 138 and 139 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69.
[0065]
　CAPRIN- having an amino acid sequence represented by SEQ ID NO: 309 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence. An amino acid or fragment thereof that is immunologically reactive with a partial polypeptide of a protein. Preferably, the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 134, 135 and 136 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 137, 138 and 139 (CDR1, CDR2 and CDR3, respectively). An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69.
[0066]
　In addition, the following anti-CAPRIN-1 antibody is also preferably used.
[0067]
　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.
[0068]
　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.
[0069]
　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.
[0070]
　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.
[0071]
　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.
[0072]
　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.
[0073]
　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.
[0074]
　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.
[0075]
　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.
[0076]
　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.
[0077]
　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.
[0078]
　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.
[0079]
　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.
[0080]
　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.
[0081]
　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.
[0082]
　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 contains the amino acid sequence of SEQ ID NO: 99.
[0083]
　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.
[0084]
　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.
[0085]
　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.
[0086]
　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.
[0087]
　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.
[0088]
　An antibody or fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111.
[0089]
　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.
[0090]
　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.
[0091]
　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.
[0092]
　An antibody or fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119.
[0093]
　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.
[0094]
　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.
[0095]
　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.
[0096]
　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.
[0097]
　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.
[0098]
　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.
[0099]
　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 contains the amino acid sequence of SEQ ID NO: 133.
[0100]
　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.
[0101]
　In the examples described later, by combining the above polyclonal antibody or monoclonal antibody against a part of the polypeptide of the region expressed on the cell membrane surface of the cancer cell and the full length of the CAPRIN-1 protein and imikimod, the strong anti-cancer thereof is achieved in the cancer-bearing organism. The tumor effect was confirmed.
[0102]
　
　The imiquimod according to the present invention is a compound represented by CAS number 99011-02-6 and having a molecular weight of about 240.3, and is an agonist that binds to Toll-like receptor (TLR) 7 or 8. The IUPAC name of imikimod is 4-amino-1- (2-methylpropyl) -1H-imidazo [4,5-c] quinoline, and as another description, R837, S-26308, 1- (2-METHYLPROPYL)- 1H-IMIDAZO [4,5-C] QUINOLIN-4-AMINE; 4-amino-1-isobutyl-1h-imidazo [4,5-c] quinoline, 1-isoButyl-1H-Inudazole [4.5-c] quinoline-4-amine; 1- (2-Methylpropyl) -1H-imidazo [4,5-c] represented by quinoline-4-amine.
[0103]
　Imiquimod may be obtained by chemical synthesis by a method known to those skilled in the art, but as a drug containing imiquimod as an active ingredient, "Veserna cream 5%" in Japan and "Aldara (registered trademark) Cream, 5%" in Europe and the United States. Is on the market, and these drugs can be appropriately used when imiquimod is used in the present invention.
[0104]
　
　The active ingredient of the drug of the present invention may contain an antitumor agent known in the literature and the like in addition to the anti-CAPRIN-1 antibody and imiquimod as long as the effect of the drug of the present invention is not impaired. The known antitumor agent is not particularly limited, and specific examples thereof include paclitaxel, doxorubicin, daunorubicin, cyclophosphamide, methotrexate, 5-fluorouracil, thiotepa, busulfan, improsulfan, piposulfan, and benzodopa. Carbocon, meturedopa, uredopa, ultretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, triethylenethiophosphamide, trimethylenethiophosphamide, trimethylenethiophosphamide, trimethylololomeramine , Briostatin, callystatin, cryptophycin 1, cryptophycin 8, drastatin, zuocalmycin, eleuterobin, pankratisstatin, sarcodictin, spongestatin, chlorambusyl, chloronaphthacil, chlorophosphamide, cholophosphamide ), Estramstin, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melfaran, novenbitin, phenesterine, prednimustine, prednimustine, prednimustine, trophosphamide, trofosphamide, trofosfamide chlorozotocin), photemustine, romustin, nimustine, ranimustin, calicheamicin, dainemycin, clodronate, esperamycin, aclassinomycin, a
[0105]
　 The combination of the anti-CAPRIN-1 antibody of the present invention and imiquimod has cytotoxic activity in vivo. Therefore, the antitumor effect of the present invention can be known by examining the cytotoxic activity against cancer. The cytotoxic activity can be evaluated by administering anti-CAPRIN-1 and imiquimod to a living body having cancer, measuring the size of the tumor after administration, and examining the size of the cancer over time. The antitumor effect of the present invention can also be 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 combination of the anti-CAPRIN-1 antibody and imiquimod in the present invention can be further known by examining the prevention of cancer, the prevention of metastasis, or the prevention of recurrence.
[0106]
　The anti-CAPRIN-1 antibody used in the present invention can be expected to have a stronger antitumor effect when the binding affinity with the CAPRIN-1 protein on the surface of cancer cells is high. 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 , and at least 5 ×. 10 9 M -1 , at least 10 10 M -1 , at least 5 x 10 10 M -1 , at least 10 11 M -1 , at least 5 x 10 11 M -1 , at least 10 12It is preferably M -1 , or at least 10 13 M -1 .
[0107]
　The ability of the anti-CAPRIN-1 antibody used in 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. it can.
[0108]
　Administration of the combination of the anti-CAPRIN-1 antibody and imiquimod in the present invention into a cancer body enhances the antitumor effect as described above, but the enhancement rate is preferably 30. % Or more, more preferably 40% or more, still more preferably 50% or more, even more preferably 55% or more, even more preferably 60% or more, even more preferably 65% ​​or more, most preferably 70% or more. The enhancement rate of the antitumor effect of the combination administration of the anti-CAPRIN-1 antibody and imiquimod in the present invention with respect to the administration of the anti-CAPRIN-1 antibody alone is such that an effective amount is administered to the cancer-bearing mouse under the same conditions, and after the start of administration, It can be calculated by comparing the tumor volumes after the 7th day.
[0109]
　 The pharmaceuticals of the present invention are intended for the treatment and / or prevention of cancer. The cancer targeted by the drug of the present invention is not particularly limited as long as it is a cancer (cell) expressing the CAPRIN-1 protein.
[0110]
　As used herein, "treatment" means the treatment of cancer based on the aforementioned antitumor effects. Further, as used herein, "prevention" means not only prevention of cancer development but also prevention of cancer metastasis or recurrence.
[0111]
　As used herein, the terms "tumor" and "cancer" mean malignant neoplasms and are used interchangeably.
[0112]
　The cancer targeted in the present invention may be any cancer as long as it expresses the CAPRIN-1 protein on the surface of the cell membrane. Preferably, the above-mentioned basal cell cancer, budget disease, skin cancer, breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, Liver cancer, bile sac cancer, sarcoma, obesity cell tumor, adrenal cortex cancer, Ewing tumor, Hodgkin lymphoma, mesenteric tumor, multiple myeloma, testicle cancer, thyroid cancer, head and neck cancer. More preferably, it is a palpable cancer, a cancer present subcutaneously, a cancer present in the skin, a superficial cancer, a cancer present in the dermis or a cancer present in a nonparenchymal organ. In addition, these cancers may be a primary cancer, a metastatic cancer, a metastatic cancer, or a recurrent cancer.
[0113]
　More specifically, the cancer includes, for example, Bowen's disease, melanoma, adenocarcinoma, extramammary Paget's disease, bacteriolytic sarcoma, Sezary syndrome, skin T / NK cell lymphoma, and T cells having lesions only in the skin. Leukemia / lymphoma, cutaneous B-cell lymphoma (independent group), cutaneous T-cell lympholine carcinoma, complex mammary adenocarcinoma, malignant mixed mammary adenocarcinoma, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, Large cell cancer, neuroepithelial tissue tumors such as adenocarcinoma, glioblastoma, neuroblastoma, ventricular lining tumor, neurocellular tumor, fetal neuroepidermal tumor, nerve sheath tumor, neurofibroma, Adenocarcinoma, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, digestive lymphoma, small to medium cell lymphoma, cecum cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, sigmoid colon cancer, Rectal cancer, ovarian epithelial cancer, embryonic cell tumor, stromal cell tumor, pancreatic duct cancer, invasive pancreatic duct cancer, adenocarcinoma of pancreatic cancer, adenocarcinoma, adenocarcinoma, giant cell tumor, papillary mucinous tumor in the pancreatic duct , Mucinous cystadenocarcinoma, pancreatoma, pancreatic head cell tumor, Frants tumor, serous cystadenocarcinoma, solid papillary cancer, gastrinoma, glucagonoma, insulinoma, multiple endocrine adenocarcinoma 1 (Wermer syndrome), non-functional island Celloma, somatostatinoma, VIP-producing tumor, cervical cancer, uterine body cancer, fibrocarcinoma, bone / articular carcinoma, Ewing's sarcoma, Wilms tumor, hepatoblastoma, soft sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, Soft malignant tumors, adenocarcinomas, and head and neck cancers include hypopharyngeal cancer, mesopharyngeal cancer, tongue cancer, nasopharyngeal cancer, oral cancer, lip cancer, sinus cancer, laryngeal cancer, and the like. Not limited to. Further, tactile cancer originating from the above cancer, cancer existing under the skin, cancer existing in the skin, superficial cancer, cancer existing in the dermis or cancer existing in a non-parenchymal organ are included. In addition, by metastasis or recurrence of the above cancer, it is present in palpable cancer, subcutaneous cancer, intradermal cancer, superficial cancer, dermis cancer or non-parenchymal organ. Cancer is included.
[0114]
　Further, the preferred subject (patient) to be a target is a mammal, for example, a mammal including a primate, a pet animal, a domestic animal, a competition animal, and the like, and a human, a dog, and a cat are particularly preferable.
[0115]
　The pharmaceutical product of the present invention can be formulated by a method known to those skilled in the art. It can be used parenterally in the form of a sterile solution, for example with water or other pharmaceutically acceptable liquid, or an injectable suspension. For example, pharmacologically acceptable carriers or vehicles, specifically sterilized water, saline, vegetable oils, emulsifiers, suspensions, surfactants, stabilizers, fragrances, excipients, binders, etc. It is conceivable to formulate by appropriately combining with and mixing in a unit dose form required for generally accepted pharmaceutical practice. The amount of active ingredient in these formulations ensures that an appropriate dose in the indicated range is available.
[0116]
　Aseptic compositions for injection can be formulated according to routine formulation practices using vehicles such as distilled water for injection. Aqueous solutions for injection include, for example, saline, isotonic solutions containing glucose and other adjuvants, such as D-sorbitol, D-mannitol, D-mannitol, sodium chloride, and suitable solubilizers such as. Alcohols, specifically ethanol, polyalcohols such as propylene glycol, polyethylene glycol and nonionic surfactants such as Polysorbate 80 (TM), HCO-60 may be used in combination. Examples of the oily liquid include sesame oil and soybean oil, and benzyl benzoate and benzyl alcohol may be used in combination as solubilizing agents. It may also be blended with buffers such as phosphate buffers, sodium acetate buffers, soothing agents such as procaine hydrochloride, stabilizers such as benzyl alcohol, phenol and antioxidants. The prepared injection solution is usually filled in a suitable ampoule. Examples of the oily liquid include sesame oil and soybean oil, and benzyl benzoate and benzyl alcohol may be used in combination as solubilizing agents. It may also be blended with buffers such as phosphate buffers, sodium acetate buffers, soothing agents such as procaine hydrochloride, stabilizers such as benzyl alcohol, phenol and antioxidants. The prepared injection solution can usually be filled in a suitable ampoule.
[0117]
　The administration is oral or parenteral, preferably parenteral administration, and specific examples thereof include an injection type, a nasal administration type, a pulmonary administration type, and a transdermal administration type. As an example of the injection type, for example, it can be administered systemically or locally by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, intratumoral injection and the like. Examples of the transdermal administration type are, for example, those called coating agents or external preparations. Examples of external preparations include solid preparations, liquid preparations, spray preparations, ointments, cream preparations, and gel preparations.
[0118]
　In addition, the administration method can be appropriately selected depending on the patient's age, weight, gender, symptoms and the like. The dose of the antibody or the pharmaceutical composition containing the polynucleotide encoding the antibody can be selected, for example, in the range of 0.0001 mg to 1000 mg per 1 kg of body weight at a time. Alternatively, the dose can be selected, for example, in the range of 0.001 to 100,000 mg / body per patient, or 1 mg to 30 mg per kg body weight of the patient, but is not necessarily limited to these numbers. The dose and administration method vary depending on the weight, age, sex, symptoms, etc. of the patient, but those skilled in the art can appropriately select the dose.
[0119]
　The effectiveness of imiquimod as a transdermal anticancer drug has been confirmed, and based on the results, imiquimod is marketed as an anticancer drug under the trade name of Aldara® (Imiquimod) Cream, 5%. As a dosage form for administering to a patient, a transdermal administration type, preferably a cream type preparation is preferable. When imiquimod is administered to a subject, it may be administered according to the dosage and administration described in the package insert of "Aldara (registered trademark) Cream, 5%".
[0120]
　
　The treatment and / or prevention of cancer by a drug for treating and / or preventing cancer of the present invention includes various forms in addition to the administration as the above-mentioned drug. For example, each active ingredient of the pharmaceutical product of the present invention can be administered simultaneously or individually in order. As a specific example, the second active ingredient can be administered within a time interval of up to about 3 weeks, that is, from immediately after the administration of the first active ingredient to about 3 weeks. At that time, the surgical procedure may be performed following the surgical procedure, or the surgical procedure may be performed between the administration of the first drug and the second drug. In addition, the therapeutic and / or prophylactic agents for cancer of the present invention may be administered according to a plurality of administration cycles. For example, when the active ingredients of the cancer treatment and / or preventive agent of the present invention are co-administered, the pharmaceutical composition containing both active ingredients is administered in a cycle of about 2 days to about 3 weeks. After that, it can be repeated as needed according to the judgment of the doctor in charge of the treatment cycle. Similarly, when planning ordered formulations, the duration of administration of each individual drug should be adjusted to span the same duration. The interval between cycles can be varied from 0 to 2 months. The dose of each active ingredient of the therapeutic and / or preventive agent for cancer of the present invention can be set in the same manner as the dose of each active ingredient in the pharmaceutical composition.
[0121]
　
　The drug for treating and / or preventing cancer of the present invention may be in the form of a pharmaceutical kit. A pharmaceutical kit is a package for using the active ingredient in the form of a separate pharmaceutical composition in a method of treating and / or preventing cancer, and the package contains instructions for administering each active ingredient. included. Each active ingredient of the above-mentioned pharmaceutical composition for treating and / or preventing cancer contained in a pharmaceutical kit is a pharmaceutical composition formulated as described above so that each active ingredient can be administered together or separately. It can be a form. In addition, the pharmaceutical kit contains a sufficient amount of the active ingredient in one or more doses so that each active ingredient can be administered according to the above-mentioned administration method.
[0122]
　Based on what is specifically described above, the present invention further provides methods for the treatment and / or prevention of cancer, including administration of the above-mentioned drug of the invention to a subject suspected of having cancer. provide. In addition, in the embodiment, the antibody or fragment thereof and the antitumor agent contained in the above-mentioned drug are administered to the above-mentioned subject simultaneously or separately.
Example
[0123]
　Hereinafter, the present invention will be specifically described based on examples, but the scope of the present invention shall not be limited by these specific examples.
[0124]
　Example 1 Preparation of anti-CAPRIN-1 antibody As the anti-CAPRIN-1 antibody
　having immunological reactivity with the CAPRIIN-1 protein used in the present invention, the one prepared as follows was used.
[0125]
　(Polyclonal antibody)
　1 mg of human CAPRIN-1 recombinant protein prepared according to Example 3 of WO2010 / 016526 was mixed with an equal volume of incomplete Freund's adjuvant (IFA) solution, and this was mixed subcutaneously with rabbits four times every two weeks. Was administered to. Blood was then collected to obtain an antiserum containing a polyclonal antibody. Further, this antiserum was purified using a protein G carrier (manufactured by GE Healthcare Bioscience) and replaced with PBS (-) to obtain a polyclonal antibody against the CAPRIN-1 protein (anti-CAPRIN-1 polyclonal antibody # 1). Obtained.
[0126]
　(
　Monclonal antibody) 100 μg of human CAPRIN-1 recombinant protein prepared according to Example 3 of WO2010 / 016526 was mixed with an equal amount of MPL + TDM adjuvant (manufactured by Sigma), and this was used as an antigen solution per mouse. The antigen solution was intraperitoneally administered to 6-week-old Balb / c mice (manufactured by Nippon SLC Co., Ltd.), and then administered 3 and 24 times every week to complete immunization. Each spleen removed 3 days after the last immunization was sandwiched between two sterilized slide glasses, ground, washed with PBS (-) (manufactured by Nissui Co., Ltd.), and centrifuged at 1500 rpm for 10 minutes to prepare the supernatant. The removal operation was repeated 3 times to obtain spleen cells. The obtained spleen cells and mouse myeloma cells SP2 / 0 (purchased from ATCC) were mixed at a ratio of 10: 1, and 200 μl of RPMI1640 medium containing 10% FBS heated to 37 ° C. and PEG1500 (Beringer). A PEG solution prepared by mixing 800 μl was added and allowed to stand for 5 minutes for cell fusion. After centrifuging at 1700 rpm for 5 minutes and removing the supernatant, cells were suspended in 150 ml of RPMI1640 medium (HAT selective medium) containing 15% FBS containing 2% equivalent of Gibco's HAT solution, and a 96-well plate (Nunk). 100 μl per well of (manufactured by) was sown on 15 plates. 7 days, 37 ° C, 5% CO 2By culturing under the conditions of (1), a hybridoma in which spleen cells and myeloma cells were fused was obtained. Hybridomas were selected based on the binding affinity of the antibody produced by the produced hybridoma to the CAPRIN-1 protein. 1 μg / ml of CAPRIN-1 protein solution was added to 1 well of a 96-well plate, and the mixture was allowed to stand at 4 ° C. for 18 hours. After washing each well with PBS-T three times, 400 μl of 0.5% Bovine Serum Albumin (BSA) solution (manufactured by Sigma) was added per well, and the mixture was allowed to stand at room temperature for 3 hours. After removing the solution, the wells were washed 3 times with 400 μl of PBS-T per well, 100 μl of each culture supernatant of the hybridoma obtained above was added per well, and the mixture was allowed to stand at room temperature for 2 hours. After washing each well with PBS-T three times, 100 μl of HRP-labeled anti-mouse IgG (H + L) antibody (manufactured by Invitrogen) diluted 5000 times with PBS was added per well and allowed to stand at room temperature for 1 hour. .. After washing the wells with PBS-T three times, 100 μl of TMB substrate solution (manufactured by Thermo) was added per well and allowed to stand for 15 to 30 minutes to carry out a color reaction. After color development, 100 μl of 1N sulfuric acid was added per well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorptiometer. As a result, a plurality of hybridomas producing an antibody having a high absorbance value were selected. The selected hybridomas were added to the plates so as to be 0.5 per well of the 96-well plate and cultured. After 1 week, hybridomas forming a single colony were observed in the wells. The cells in these wells were further cultured, and hybridomas were selected based on the binding affinity of the antibody produced by the cloned hybridoma to the CAPRIN-1 protein. 1 μg / ml of CAPRIN-1 protein solution was added to 1 well of a 96-well plate, and the mixture was allowed to stand at 4 ° C. for 18 hours. After washing each well with PBS-T three times, add 400 μl of 0.5% BSA solution per well. The mixture was allowed to stand at room temperature for 3 hours. After removing the solution, the wells were washed 3 times with 400 μl of PBS-T per well, 100 μl of each culture supernatant of the hybridoma obtained above was added per well, and the mixture was allowed to stand at room temperature for 2 hours. After washing each well with PBS-T three times, 100 μl of HRP-labeled anti-mouse IgG (H + L) antibody (manufactured by Invitrogen) diluted 5000-fold with PBS was added per well and allowed to stand at room temperature for 1 hour. After washing the wells with PBS-T three times, 100 μl of a TMB substrate solution (manufactured by Thermo) was added per well and allowed to stand for 15 to 30 minutes for a color reaction. After color development, 100 μl of 1N sulfuric acid was added per well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorptiometer. As a result, a plurality of mouse monoclonal antibodies showing reactivity with the CAPRIN-1 protein were obtained.
[0127]
　Furthermore, by the flow cytometry method, the reactivity to human cancer cells in which the CAPRIN-1 protein was confirmed to be expressed on the cell membrane surface was confirmed. As a negative control, a mouse IgG control antibody showing no reactivity with the cancer cells was used. As a result of confirmation, we obtained several monoclonal antibodies that have stronger fluorescence intensity than the mouse IgG control antibody and strongly react with the cell membrane surface of the cancer cells expressing CAPRIN-1 on the cell membrane surface. It was. Among them, as a monoclonal antibody exhibiting reactivity with the CAPRIN-1 protein, it is a monoclonal antibody against CAPRIN-1 described in WO2013 / 125630, and the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 114. , An antibody containing the amino acid sequence of the light chain variable region represented by SEQ ID NO: 115 was selected.
[0128]
　The CDRs 1 to 3 of the heavy chain variable region of the selected antibody were identified, and the base sequence was designed so that the framework region could express the heavy chain variable region containing the human antibody sequence, and this was determined as the heavy chain of human IgG1. It was inserted into a mammalian expression vector into which a normal region was inserted. Similarly, CDRs 1 to 3 of the light chain variable region were identified, and the base sequence was designed so that the framework region could express the light chain variable region containing the sequence of the human antibody, and this was used as the light chain constant of human IgG1. It was inserted into a mammalian expression vector into which the region was inserted. The above two recombinant expression vectors were introduced into mammalian cells according to a conventional method to obtain a culture supernatant containing a humanized monoclonal antibody # 1 (humanized antibody # 1) against CAPRIN-1.
[0129]
　The obtained culture supernatant containing the humanized anti-CAPRIN-1 monoclonal antibody # 1 was purified using Hitrap Protein A SepharoseFF (manufactured by GE Healthcare) according to a conventional method, replaced with PBS (-), and 0.22 μm. A filter (manufactured by Millipore) was prepared.
[0130]
　The specific reactivity of the anti-CAPRIN-1 antibody with the CAPRIN-1 protein was confirmed by solidifying the CAPRIN-1 protein on a plate and detecting it using the ELISA method.
[0131]
　By examining the reactivity with cancer cells that have not undergone permeation treatment of the cell membrane by the flow cytometry method using the above anti-CAPRIN-1 antibody, as shown in the following examples, a part of CAPRIN-1. Was confirmed to be expressed on the cell membrane surface of cancer cells.
[0132]
　Human cancer cells whose expression of the CAPRIN-1 gene has been confirmed by the flow cytometry method: breast cancer cells (BT-474), colon cancer cells (HT-29), lung cancer cells (QG56, H1650), gastric cancer Cells (NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal cancer Cells (Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), bile sac cancer cells (TGBC14TKB), For any of the cancer cells of fibrosarcoma cells (HT-1080), melanoma cells (G-361), mouse renal cancer cells (Renca) and mouse breast cancer cells (4T1) whose expression of the CAPRIN-1 gene has been confirmed. However, humanized antibody # 1 has stronger fluorescence intensity than human IgG control antibody and rabbit IgG antibody, which are not reactive with cancer cells, which are negative controls, and the above cancer cells expressing CAPRIN-1. It was confirmed that it reacts strongly with the cell membrane surface of.
[0133]
　Example 2 Anti-tumor effect of the combined use of anti-CAPRIN-1 antibody and imiquimod
　Next, the anti-CAPRIN-1 antibody (anti-CAPRIN-1 polyclonal antibody # 1 and humanized antibody # 1) and imiquimod prepared in Example 1 The anti-tumor effect in vivo of cancer-bearing mice was evaluated by administration in combination with.
[0134]
　Specifically, using NOD-SCID mice in which human-derived cancer cells expressing the CAPRIN-1 protein are subcutaneously transplanted, the antitumor effect of the combined use of the anti-CAPRIN-1 antibody and imiquimod in the present invention can be obtained. investigated. 10 per mouse 7 a single human breast cancer cells BT474 were mixed with Matrigel (SIGMA) was subcutaneously transplanted tumor about 60 mm 3 were produced tumor-bearing mice were grown to. BT474 is a cancer cell in which the CAPRIN-1 protein is expressed on the cell membrane surface, and it has been confirmed that the anti-CAPRIN-1 antibody prepared in Example 1 reacts with a part of CAPRIN-1 on the cell membrane surface. .. The anti-CAPLIN-1 antibody prepared in Example 1 was administered to the tail veins of the above-mentioned cancer-bearing mice at 10 mg / kg once a week. Imiquimod was started to be applied to the same mice at the same time as the first administration of anti-CAPRIN-1 antibody. "Versena Cream 5%" containing imiquimod as an active ingredient (Mochida Pharmaceutical Co., Ltd., hereinafter referred to as "imiquimod cream") is applied to the surface of the skin on which cancer cells have been transplanted for 5 consecutive days a week, and then. No administration was performed for 2 days, and the same antibody administration and imiquimod cream as above were applied on the 8th day from the start of antibody administration.
[0135]
　As a control group, the same anti-CAPRIN-1 antibody as above was administered to cancer-bearing mice at the same dose once a week. Further, as a control group, imiquimod cream was applied to another cancer-bearing mouse individual on the surface of the skin into which the cancer cells were transplanted at similar administration intervals. Furthermore, cancer-bearing mice in the untreated group were used as negative controls. After the start of administration, the size of the cancer in the cancer-bearing mouse was measured over time using a caliper, and the tumor volume was calculated according to the formula: (length of the major axis of the cancer) × (length of the minor axis of the cancer). ) Calculated as 2 x 0.5.
[0136]
　As a result of the evaluation, on the 45th day after the administration of the anti-CAPRIN-1 polyclonal antibody # 1 was started, when the tumor volume of the negative control was set to 100%, the anti-CAPRIN-1 polyclonal antibody # 1 prepared in Example 1 and the anti-CAPRIN-1 polyclonal antibody # 1 and It was less than 60% in the cancer-bearing mice to which the imiquimod cream was administered. Among the cancer-bearing mice in the control group, 78% were administered with anti-CAPRIN-1 polyclonal antibody # 1 alone, and 69% were administered with imiquimod cream alone.
[0137]
　On the 48th day after the start of administration of the humanized antibody # 1 which is an anti-CAPRIN-1 monoclonal antibody, when the tumor volume of the negative control was set to 100%, the humanized antibody # 1 prepared in Example 1 and the imiquimod cream were prepared. It was less than 33% in the cancer-bearing mice that received. In the cancer-bearing mice of the comparative control group, 65% was administered only with humanized antibody # 1. It was 69% in the group receiving only imiquimod cream.
[0138]
　As a result of this evaluation, administration of the antibody against CAPRIN-1 and imiquimod in combination has a significantly stronger antitumor effect than the case where the anti-CAPRIN-1 antibody is administered alone and the case where imiquimod is administered alone. Was shown.
[0139]
　Example 3 Comparison of the antitumor effect of the combined use of the anti-CAPRIN-1 antibody and the anti-cancer agent
　Next, the anti-CAPRIN-1 antibody (anti-CAPRIN-1 polyclonal antibody # 1 and humanized antibody # 1) prepared in Example 1 ) And imikimod were administered in combination to evaluate the antitumor effect in vivo of cancer-bearing mice and the antitumor effect when the above antibody was used in combination with an existing anticancer agent different from imikimod.
[0140]
　Specifically, using NOD-SCID mice in which human-derived cancer cells expressing the CAPRIN-1 protein are subcutaneously transplanted, the antitumor effect of the combined use of the anti-CAPRIN-1 antibody and imiquimod in the present invention can be obtained. investigated. 10 per mouse 7 a single human breast cancer cells BT474 were mixed with Matrigel (SIGMA) was subcutaneously transplanted tumor about 90 mm 3 were produced tumor-bearing mice were grown to. BT474 is a cancer cell in which the CAPRIN-1 protein is expressed on the cell membrane surface, and the anti-CAPLIN-1 antibody prepared in Example 1 is confirmed to react with a part of CAPRIN-1 on the cell membrane surface. ..
[0141]
　In the combination treatment group of anti-CAPRIN-1 antibody and imiquimod, 10 mg / kg of each of the anti-CAPRIN-1 antibody (humanized antibody # 1) prepared in Example 1 was added to the tail vein of the above-mentioned cancer-bearing mouse. It was administered once a week. Furthermore, imiquimod was applied to the skin surface of the site where the cancer was present at the same time as the first administration of the anti-CAPRIN-1 antibody to the same mouse. "Versena Cream 5%" (Mochida Pharmaceutical Co., Ltd., hereinafter referred to as "imiquimod cream") contained as an active ingredient of imiquimod is applied to the surface of the skin on which cancer cells are transplanted for 5 consecutive days a week, and then 2 No administration was performed for the day, and on the 8th day after the start of antibody administration, the same antibody administration and imiquimod cream as above were applied.
[0142]
　In the combined treatment group of the anti-CAPRIN-1 antibody and the anti-cancer agent, 10 animals of each of the prepared anti-CAPRIN-1 antibody (humanized antibody # 1) prepared in Example 1 were added to the tail vein of the prepared cancer-bearing mouse at 10 mg / kg. It was administered once a week. Further, the same mice were intraperitoneally administered with the anticancer agents paclitaxel and docetaxel at 7 mg / kg and 8 mg / kg once a week at the same time as the initial administration of the anti-CAPRIN-1 antibody.
[0143]
　As a control group, the same anti-CAPRIN-1 antibody as above was administered to cancer-bearing mice at the same dose once a week. After the start of administration, the size of the cancer in the cancer-bearing mouse was measured over time using a caliper, and the tumor volume was calculated according to the formula: (length of the major axis of the cancer) × (length of the minor axis of the cancer). ) Calculated as 2 x 0.5.
[0144]
　As a result of comparative evaluation, on the 53rd day after the start of administration of humanized antibody # 1, when the tumor volume of the mouse to which only the anti-CAPRIN-1 antibody was administered was 100%, the anti-CAPRIN- prepared in Example 1 was prepared. It was less than 20% in the cancer-bearing mice to which 1 antibody and imiquimod cream were administered. On the other hand, when paclitaxel and docetaxel were used in combination with humanized antibody # 1, the percentage was 47% or more.
[0145]
　Similarly, anti-CAPRIN-1 polyclonal antibody # 1 was subjected to the same comparative evaluation as above. As a result, on the 30th day after the start of administration of anti-CAPRIN-1 polyclonal antibody # 1, the anti-CAPRIN-1 polyclonal antibody # 1 was started. When the tumor volume of the mouse to which only # 1 was administered was 100%, it was less than 46% of the cancer-bearing mouse to which the anti-CAPRIN-1 polyclonal antibody # 1 and imiquimod cream were co-administered. On the other hand, when paclitaxel and docetaxel were used in combination with anti-CAPRIN-1 polyclonal antibody # 1, the ratio was 65% or more.
[0146]
　As a result of this evaluation, the antitumor effect when the antibody against CAPRIN-1 and imiquimod were treated in combination was compared with the antitumor effect when the antibody against CAPRIN-1 and an existing anticancer agent different from imiquimod were treated in combination. It has been shown to have a remarkably strong synergistic antitumor effect.
The scope of the claims
[Claim 1]
　A pharmaceutical product for the treatment and / or prevention of cancer, which comprises an antibody or fragment thereof having immunological reactivity with the CAPRIN-1 protein and imiquimod in combination or separately.
[Claim 2]
　The antibody or fragment is a CAPRIN-1 protein having an amino acid sequence represented by any of SEQ ID NOs: 2 to 30, which is an even number, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. The drug according to claim 1, which is an antibody or a fragment thereof having immunological reactivity.
[Claim 3]
　The pharmaceutical product according to claim 1 or 2, wherein the antibody or fragment thereof has immunological reactivity with the extracellular region of the CAPRIN-1 protein present on the surface of cancer cells.
[Claim 4]
　CAPRIN-in which 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 drug according to any one of claims 1 to 3, which has immunological reactivity with a partial polypeptide of one protein.
[Claim 5]
　The drug according to any one of claims 1 to 4, wherein the antibody is a monoclonal antibody or a polyclonal antibody.
[Claim 6]
　The drug according to any one of claims 1 to 5, wherein the antibody or fragment thereof is any of the following (A) to (M).
(A) Heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 36, 37 and 38 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 40, 41 and 42 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments (B) SEQ ID NOs: 44, 45 and 46 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable regions and are immunologically reactive with the CAPRIN-1 protein. Contains heavy chain variable regions containing, and light chain variable regions containing complementarity determining regions of SEQ ID NOs: 48, 49 and 50 (CDR1, CDR2 and CDR3, respectively), and immunoreactivity with CAPRIN-1 proteins. The
heavy chain variable regions containing the complementarity determining regions of antibody or fragment (C) SEQ ID NOs: 52, 53 and 54 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 56, 57 and 58 (CDR1, respectively).
The complementarity determining regions of antibodies or fragments (D) SEQ ID NOs: 60, 61 and 62 (CDR1, respectively, ) comprising a light chain variable region containing CDR2 and CDR3) and immunoreactive with the CAPRIN-1 protein . It contains heavy chain variable regions containing CDR2 and CDR3) and light chain variable regions containing complementarity determining regions of SEQ ID NOs: 64, 65 and 66 (CDR1, CDR2 and CDR3, respectively), and CAPRIN-1 protein and immunology. Reactive antibodies or fragments
(E) Heavy chain variable regions containing complementarity determining regions 170, 171 and 172 (CDR1, CDR2 and CDR3 respectively) and complementarity determining regions of SEQ ID NOs: 173, 174 and 175 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments thereof (F) SEQ ID NOs: 176, 177 and 178 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable region and have immunoreactivity with the CAPRIN-1 protein. ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 (CDR1, CDR2 and CDR3, respectively) and immunoreactivity with the CAPRIN-1 protein. Antibodies or fragments thereof
(G) with heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 182, 183 and 184 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 185, 186 and 187, respectively.
The complementarity determining regions (H) SEQ ID NOs: 188, 189 and 190 of an antibody or fragment (H) thereof that contains light chain variable regions containing CDR1, CDR2 and CDR3) and is immunologically reactive with the CAPRIN-1 protein. The heavy chain variable regions containing CDR1, CDR2 and CDR3) and the light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively), and the CAPRIN-1 protein. Antibodies or fragments thereof that are immunologically reactive with
(I) Heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 146, 147 and 148 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 149, 150 and 151 (CDR1, CDR2 and CDR3, respectively).
The complementarity determining regions of antibodies or fragments thereof (J) SEQ ID NOs: 272, 273 and 274 (CDR1, CDR2 and CDR3, respectively) that contain the light chain variable region containing the CAPRIN-1 protein and have immunoreactivity with the CAPRIN-1 protein. ) And light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 275, 276 and 277 (CDR1, CDR2 and CDR3, respectively) and immunoreactivity with the CAPRIN-1 protein. Antibodies or fragments thereof
(K) with heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 290, 291 and 292 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 293, 294 and 295, respectively.
The complementarity determining regions (L) of the antibody or fragment (L) SEQ ID NOs: 301, 302 and 303 containing a light chain variable region containing CDR1, CDR2 and CDR3) and immunoreactive with the CAPRIN-1 protein. The heavy chain variable regions containing CDR1, CDR2 and CDR3) and the light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 305, 306 and 307 (CDR1, CDR2 and CDR3, respectively), and the CAPRIN-1 protein. Antibodies or fragments thereof that are immunologically reactive with
(M) Heavy chain variable regions containing complementarity determining regions of SEQ ID NOs: 134, 135 and 136 (CDR1, CDR2 and CDR3, respectively) and complementarity determining regions of SEQ ID NOs: 137, 138 and 139 (CDR1, CDR2 and CDR3, respectively). Antibodies or fragments thereof that contain light chain variable regions containing, and have immunological reactivity with CAPRIN-1 proteins.
[Claim 7]
　The drug according to any one of claims 1 to 6, wherein the antibody or fragment thereof is any one of the following (a) to (ak).
(A) An antibody or a fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 39 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 43. (B) The heavy chain variable region of SEQ ID NO: 47. An antibody or fragment thereof comprising an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 51
(c) The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 55 and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 59
(d) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 63 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 67. An
antibody in which the fragment (e) heavy chain variable region contains the amino acid sequence of SEQ ID NO: 68 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 69 or a fragment
(f) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(g) containing 70 amino acid sequences and the light chain variable region containing the amino acid sequence of SEQ ID NO: 71. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 72 and the light chain variable. An antibody or fragment thereof
(h) in which the region comprises the amino acid sequence of SEQ ID NO: 73, 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. Antibody or fragment thereof
(I) An antibody or fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 76 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 77. (J) The heavy chain variable region of SEQ ID NO: 78. An antibody or fragment thereof
(k) containing an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 79. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 80, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 81
(l) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 82 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 83. An
antibody in which the fragment (m) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 85 or a fragment
(n) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(o) containing the amino acid sequence of 86 and the light chain variable region containing the amino acid sequence of SEQ ID NO: 87. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 88 and the light chain variable. An antibody or fragment thereof
(p) in which the region comprises the amino acid sequence of SEQ ID NO: 89 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. Antibody or fragment thereof
(q) An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 92 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 93.
(R) An antibody or fragment thereof in which the
heavy chain variable region contains the amino acid sequence of SEQ ID NO: 94 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 95. (S) The heavy chain variable region of SEQ ID NO: 96. An antibody or fragment thereof
(t) containing an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 97. The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 98, and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 99.
(U) An antibody or an antibody in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 100 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 101. The fragment
(v) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 103, or the fragment
(w) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(x) containing 104 amino acid sequences and the light chain variable region containing the amino acid sequence of SEQ ID NO: 105 contains the amino acid sequence of SEQ ID NO: 106 and the light chain variable region An antibody or fragment thereof
(y) in which the region comprises the amino acid sequence of SEQ ID NO: 107. 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. Antibody or fragment thereof
(z) An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 110 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 111.
(Aa) An antibody or fragment thereof
(ab) in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 113 (ab) The heavy chain variable region of SEQ ID NO: 114. An antibody or fragment
(ac) thereof comprising an amino acid sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 115 The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 116, and the light chain variable region An antibody comprising the amino acid sequence of SEQ ID NO: 117 or a fragment thereof
(ad) An antibody or an antibody in which 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. An
antibody in which the fragment (ae) heavy chain variable region contains the amino acid sequence of SEQ ID NO: 120 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 121 or a fragment
(af) heavy chain variable region thereof is SEQ ID NO: An antibody or fragment thereof
(ag) containing the amino acid sequence of 122 and having the light chain variable region containing the amino acid sequence of SEQ ID NO: 123 The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 124 and the light chain variable The antibody or fragment
(ah) heavy chain variable region comprising the region comprising the amino acid sequence of SEQ ID NO: 125 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. Antibody or fragment thereof
(ai) An antibody or fragment thereof comprising a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 128 and a light chain variable region containing the amino acid sequence of SEQ ID NO: 129.
(Ai) An antibody in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 131 or a fragment thereof
(aj) The heavy chain variable region of SEQ ID NO: 132 An antibody comprising an amino acid sequence and the light chain variable region contains the amino acid sequence of SEQ ID NO: 133 or a fragment
(ak) thereof The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 300 and the light chain variable region An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 304.
[Claim 8]
　The pharmaceutical product according to any one of claims 1 to 7, wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single chain antibody.
[Claim 9]
　The drug according to any one of claims 1 to 8, wherein the cancer is a cancer that expresses the CAPRIN-1 protein on the cell membrane surface.
[Claim 10]
　The cancers are basal cell cancer, budget disease, skin cancer, breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain tumor, gastric cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, Any of claims 1 to 9, which is liver cancer, bile sac cancer, sarcoma, obesity cell tumor, adrenal cortex cancer, Ewing tumor, Hodgkin lymphoma, mesenteric tumor, multiple myeloma, testicular cancer, thyroid cancer or head and neck cancer. Or the drug described in item 1.
[Claim 11]
　The drug according to any one of claims 1 to 10, wherein the dosage form of imiquimod is a transdermal preparation.
[Claim 12]
　An agent for enhancing the efficacy of a pharmaceutical composition for treating and / or preventing cancer, which comprises an antibody or a fragment thereof having immunological reactivity with the CAPRIN-1 protein as an active ingredient and containing imiquimod as an active ingredient.
[Claim 13]
　An agent for enhancing the efficacy of a pharmaceutical composition for treating and / or preventing cancer containing imiquimod as an active ingredient, which comprises an antibody having immunological reactivity with the CAPRIN-1 protein or a fragment thereof.
[Claim 14]
　A method for the treatment and / or prevention of cancer, characterized in that an antibody or fragment thereof having immunological reactivity with the CAPRIN-1 protein and imiquimod are administered to a subject together or separately.