Medicine for treating and / or preventing cancer

Abstract

A drug comprising together or separately a combination of an antibody or a fragment thereof having immunoreactivity with a CAPRIN-1 protein and a RET kinase inhibitor is useful for the treatment and / or prevention of cancer.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical for treating and / or preventing cancer using an antibody against a CAPRIN-1 protein or a fragment thereof and a RET kinase inhibitor. Background Art

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

[0003] In recent years, in order to improve the effectiveness of cancer therapeutic drugs clinically, a treatment method combining multiple cancer therapeutic drugs has been used as a standard treatment method. For example, a treatment method combining doxorubicin and cyclophosphamide, or a treatment combining multiple anticancer agents such as paclitaxel, trastuzumab and pertuzumab has become a general method. For cancer therapeutic drugs with anti-CAPRIN-1 antibodies as active ingredients, excellent cancer therapeutic effects have been confirmed by combining with chemotherapeutic agents (Patent Document 2). However, the treatment of cancer using a combination of chemotherapeutic agents is not effective for all cancers used. In addition, although the therapeutic effect is sometimes improved cumulatively, the number of synergistically significantly improving the therapeutic effect is relatively small.

[0004] In addition, in recent years, the development of molecular target therapeutic drugs has been vigorously carried out as cancer therapeutic drugs, and one of the targets that has attracted attention is RET (Rearranged During Transfection) kinase. RET kinase is a receptor tyrosine kinase that binds to the extracellular signal transduction molecules of the GDNF (glial cell line-derived neurotrophic factor) family and is encoded by the RET gene in humans. It has been found that the fusion RET gene is detected in a part of lung cancer and is also involved in the onset of lung cancer (non-patent document 1). The so-called RET fusion gene is known to be a gene located on chromosome 10. The gene is activated by being cut off in the middle and fused with partner genes such as KIF5B and CCDC6 to generate abnormal proteins. It has been confirmed that RET fusion exists not only in lung cancer, but also in multiple cancer types. For lung cancer positive for RET fusion gene, it shows the possibility that molecular target therapeutic drugs that inhibit RET are effective, and the development of new molecular target treatments for RET lung cancer patients has attracted attention (non-patent document 2). In addition, it is reported that even lung cancer that has become resistant to RET inhibitors by acquiring Gatekeeper mutations (gene mutations generated at the binding site of the inhibitor) has a therapeutic effect with the next-generation RET inhibitor selpatinib, and is expected to have promising effects on lung cancer treatment (Non-Patent Document 3). However, the acquisition of resistance to molecular target therapeutic drugs by cancer cells due to secondary mutations that confer resistance on the activation loop that is located away from the binding site of the drug in the RET fusion protein has become a major obstacle to the therapeutic effect (Non-Patent Document 4).

[0005] Prior art literature

[0006] Patent Literature

[0007] Patent document 1: WO2010 / 016526

[0008] Patent Document 2: WO2011 / 096535

[0009] Non-patent literature

[0010] Non-patent literature 1: Nature Medicine, 2012, 18, 375-377.

[0011] Non-patent literature 2: Lancet Respir Med. 2017, 5(1): 42-50.

[0012] Non-patent literature 3: JCO Precis Oncol. 2019, 3: PO.19.00189.

[0013] Non-patent document 4: Nat Commun. 2018, 9(1): 625. Summary of the invention

[0014] Problems to be solved by the invention

[0015] An object of the present invention is to provide a pharmaceutical for treating and / or preventing cancers that specifically express CAPRIN-1 protein on the cell surface.

[0016] Means for solving problems

[0017] As a result of intensive studies, the present inventors have found that the combination of an antibody against CAPRIN-1 protein or a fragment thereof having immunoreactivity with cancer cells and a RET kinase inhibitor exerts a very strong anti-tumor effect, thereby completing the present invention.

[0018] Specifically, the present invention relates to the following embodiments (1) to (14).

[0019] (1) A pharmaceutical for treating and / or preventing cancer, comprising, together or separately, an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein and a RET (Rearranged During Transfection) kinase inhibitor.

[0020] (2) The drug according to (1), wherein the RET kinase inhibitor is selpercatinib.

[0021] (3) The drug according to (1) or (2), wherein the antibody or fragment thereof is immunoreactive with a CAPRIN-1 protein having an amino acid sequence represented by any of even-numbered sequence numbers 2 to 30, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence.

[0022] (4) The pharmaceutical according to any one of (1) to (3), wherein the antibody or fragment thereof is immunoreactive with the extracellular region of the CAPRIN-1 protein present on the surface of cancer cells.

[0023] (5) The pharmaceutical according to any one of (1) to (4), wherein the antibody or fragment thereof is immunoreactive with a partial polypeptide of the CAPRIN-1 protein having an amino acid sequence as shown in any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, or 309, or an amino acid sequence having a sequence identity of 80% or more to the amino acid sequence.

[0024] (6) The pharmaceutical according to any one of (1) to (5), wherein the antibody is a monoclonal antibody or a polyclonal antibody.

[0025] (7) The pharmaceutical according to any one of (1) to (6), wherein the antibody or fragment thereof is any one of the following (A) to (M):

[0026] (A) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 36, a complementarity determining region CDR2 set forth in SEQ ID NO: 37, and a complementarity determining region CDR3 set forth in SEQ ID NO: 38, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 40, a complementarity determining region CDR2 set forth in SEQ ID NO: 41, and a complementarity determining region CDR3 set forth in SEQ ID NO: 42;

[0027] (B) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 44, a complementarity determining region CDR2 set forth in SEQ ID NO: 45, and a complementarity determining region CDR3 set forth in SEQ ID NO: 46, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 48, a complementarity determining region CDR2 set forth in SEQ ID NO: 49, and a complementarity determining region CDR3 set forth in SEQ ID NO: 50;

[0028] (C) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 52, a complementarity determining region CDR2 set forth in SEQ ID NO: 53, and a complementarity determining region CDR3 set forth in SEQ ID NO: 54, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 56, a complementarity determining region CDR2 set forth in SEQ ID NO: 57, and a complementarity determining region CDR3 set forth in SEQ ID NO: 58;

[0029] (D) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 60, a complementarity determining region CDR2 set forth in SEQ ID NO: 61, and a complementarity determining region CDR3 set forth in SEQ ID NO: 62, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 64, a complementarity determining region CDR2 set forth in SEQ ID NO: 65, and a complementarity determining region CDR3 set forth in SEQ ID NO: 66;

[0030] (E) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 170, a complementarity determining region CDR2 set forth in SEQ ID NO: 171, and a complementarity determining region CDR3 set forth in SEQ ID NO: 172, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 173, a complementarity determining region CDR2 set forth in SEQ ID NO: 174, and a complementarity determining region CDR3 set forth in SEQ ID NO: 175;

[0031] (F) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 176, a complementarity determining region CDR2 set forth in SEQ ID NO: 177, and a complementarity determining region CDR3 set forth in SEQ ID NO: 178, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 179, a complementarity determining region CDR2 set forth in SEQ ID NO: 180, and a complementarity determining region CDR3 set forth in SEQ ID NO: 181;

[0032] (G) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 182, a complementarity determining region CDR2 set forth in SEQ ID NO: 183, and a complementarity determining region CDR3 set forth in SEQ ID NO: 184, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 185, a complementarity determining region CDR2 set forth in SEQ ID NO: 186, and a complementarity determining region CDR3 set forth in SEQ ID NO: 187;

[0033] (H) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 188, a complementarity determining region CDR2 set forth in SEQ ID NO: 189, and a complementarity determining region CDR3 set forth in SEQ ID NO: 190, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 191, a complementarity determining region CDR2 set forth in SEQ ID NO: 192, and a complementarity determining region CDR3 set forth in SEQ ID NO: 193;

[0034] (I) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 146, a complementarity determining region CDR2 set forth in SEQ ID NO: 147, and a complementarity determining region CDR3 set forth in SEQ ID NO: 148, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 149, a complementarity determining region CDR2 set forth in SEQ ID NO: 150, and a complementarity determining region CDR3 set forth in SEQ ID NO: 151;

[0035] (J) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 272, a complementarity determining region CDR2 set forth in SEQ ID NO: 273, and a complementarity determining region CDR3 set forth in SEQ ID NO: 274, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 275, a complementarity determining region CDR2 set forth in SEQ ID NO: 276, and a complementarity determining region CDR3 set forth in SEQ ID NO: 277;

[0036] (K) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 290, a complementarity determining region CDR2 set forth in SEQ ID NO: 291, and a complementarity determining region CDR3 set forth in SEQ ID NO: 292, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 293, a complementarity determining region CDR2 set forth in SEQ ID NO: 294, and a complementarity determining region CDR3 set forth in SEQ ID NO: 295;

[0037] (L) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 301, a complementarity determining region CDR2 set forth in SEQ ID NO: 302, and a complementarity determining region CDR3 set forth in SEQ ID NO: 303, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 305, a complementarity determining region CDR2 set forth in SEQ ID NO: 306, and a complementarity determining region CDR3 set forth in SEQ ID NO: 307;

[0038] (M) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 134, a complementarity determining region CDR2 set forth in SEQ ID NO: 135, and a complementarity determining region CDR3 set forth in SEQ ID NO: 136, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 137, a complementarity determining region CDR2 set forth in SEQ ID NO: 138, and a complementarity determining region CDR3 set forth in SEQ ID NO: 139.

[0039] (8) The pharmaceutical according to any one of (1) to (7), wherein the antibody or fragment thereof is any one of the following (a) to (a1),

[0040] (a) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43;

[0041] (b) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:47 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:51;

[0042] (c) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59;

[0043] (d) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 67;

[0044] (e) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69;

[0045] (f) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71;

[0046] (g) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73;

[0047] (h) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75;

[0048] (i) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77;

[0049] (j) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79;

[0050] (k) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81;

[0051] (l) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83;

[0052] (m) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85;

[0053] (n) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87;

[0054] (o) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89;

[0055] (p) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 91;

[0056] (q) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:93;

[0057] (r) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:95;

[0058] (s) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:97;

[0059] (t) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 99;

[0060] (u) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101;

[0061] (v) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103;

[0062] (w) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105;

[0063] (x) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107;

[0064] (y) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109;

[0065] (z) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111;

[0066] (aa) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113;

[0067] (ab) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115;

[0068] (ac) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117;

[0069] (ad) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119;

[0070] (ae) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121;

[0071] (af) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123;

[0072] (ag) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125;

[0073] (ah) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127;

[0074] (ai) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129;

[0075] (aj) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131;

[0076] (ak) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133;

[0077] (a1) An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 304.

[0078] (9) The pharmaceutical according to any one of (1) to (8), wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single-chain antibody.

[0079] (10) The pharmaceutical according to any one of (1) to (9), wherein the cancer is a cancer expressing CAPRIN-1 protein on the cell membrane surface.

[0080] (11) The drug according to any one of (1) to (10), wherein the cancer is colorectal cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, kidney cancer, breast cancer, gastric cancer, bile duct cancer, thyroid cancer, melanoma, renal cell carcinoma, Hodgkin lymphoma, head and neck cancer, mesothelioma, colon / rectal cancer, esophageal cancer, esophagogastric junction cancer, hepatocellular carcinoma, glioblastoma, urothelial carcinoma, bladder cancer, uterine cancer, primary central nervous system lymphoma, primary testicular lymphoma, biliary tract cancer, brain tumor, leukemia, liver cancer, sarcoma, fibrosarcoma, mast cell tumor, adrenocortical carcinoma, Ewing sarcoma, testicular cancer, basal cell carcinoma, lymphoma, multiple myeloma, Paget's disease or skin cancer.

[0081] (12) A drug efficacy enhancer for a pharmaceutical composition for treating and / or preventing cancer, wherein the pharmaceutical composition for treating and / or preventing cancer comprises an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein as an active ingredient, and wherein the drug efficacy enhancer comprises a RET kinase inhibitor as an active ingredient.

[0082] (13) A drug efficacy enhancer for a pharmaceutical composition for treating and / or preventing cancer, wherein the pharmaceutical composition for treating and / or preventing cancer comprises a RET kinase inhibitor as an active ingredient, and wherein the drug efficacy enhancer comprises an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein as an active ingredient.

[0083] (14) A method for treating and / or preventing cancer, characterized in that an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein and a RET kinase inhibitor are administered to a subject together or separately.

[0084] Effects of the Invention

[0085] The combination of the antibody against CAPRIN-1 protein or its fragment and RET kinase inhibitor according to the present invention exerts a stronger anti-tumor effect than the antibody against CAPRIN-1 protein alone or the existing chemotherapeutic agent alone. In particular, although the signal transduction system that is the target of RET kinase inhibitors is not directly related to the immune system, the present invention can achieve the effect of enhancing the phagocytic activity of cancer cells by immune cells. Therefore, the combination of the antibody against CAPRIN-1 protein or its fragment and RET kinase inhibitor is effective for the treatment and prevention of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Figure 1This is a graph showing the phagocytic activity of the combination of anti-CAPRIN-1 antibody and selpatinib on a human colorectal cancer cell line (HCT116) via human monocytes (THP-1). Reference number 1: Combination drug non-addition test area. Reference number 2: Serpatinib combination test area (5μM). The bar graph shows the average value of the data obtained for each sample with N=3. The error bar represents the standard deviation (SD). The result of the Student's t-test showed that the phagocytic activity of the drug combination test area was significantly higher than that of the combination drug non-addition test area (p<0.001; significance level 5%).

[0087] Figure 2 This is a graph showing the phagocytic activity of the combination of anti-CAPRIN-1 antibody and selpatinib on the human colorectal cancer cell line (HCT116) via human monocytes (THP-1). Reference number 1: Combination drug non-addition test area. Reference number 2: Serpatinib combination test area (5μM). Reference number A: Control IgG antibody treatment group; Reference number B: Anti-CAPRIN-1 antibody treatment group. The bar graph shows the average value of the data obtained for each sample with N=3. The error bar represents the standard deviation (SD). The result of Dunnett's test showed that in the anti-CAPRIN-1 antibody treatment group, the phagocytic activity of the drug combination test area was significantly higher than that of the combination drug non-addition test area (p<0.001; significance level 5%).

[0088] Figure 3 It is a graph showing the phagocytic activity of anti-CAPRIN-1 antibody, anti-CD20 antibody and anti-HER-2 antibody alone or in combination with selpercatinib on human colorectal cancer cell line (HCT116) via human monocytes (THP-1). Reference number 3: Combination drug non-addition test area. Reference number 4: Selpercatinib combination test area (5μM). A: Anti-CAPRIN-1 antibody test area (1μM), B: Anti-CD20 antibody test area (1μM), C: Anti-HER-2 antibody test area (1μM). The bar graph shows the average value of data obtained with N=3 for each sample. The error bar represents the standard deviation (SD). As a result of Dunnett's test, in the anti-CAPRIN-1 antibody test area, the phagocytic activity of the drug combination test area was significantly higher than that of the combination drug non-addition test area (p<0.001; significance level 5%). In the anti-CD20 antibody or anti-HER-2 antibody test area, no significant changes were observed in the phagocytic activity of the drug combination test area compared with the drug combination non-addition test area. ns: not significant.

[0089] Figure 4It is a graph showing the phagocytic activity of anti-CAPRIN-1 antibody alone or in combination with selpercatinib or cisplatin on human colorectal cancer cell line (HCT116) via human monocytes (THP-1). Reference number 5: test area without addition of combined drug. Reference number 6: selpercatinib (5μM), reference number 7: test area with cisplatin combination (1μM). The bar graph shows the mean value of data obtained for each sample with N=3. The error bar indicates the standard deviation (SD). Test: Dunnett's test, p<0.001, significance level 5%. ns: not significant. DETAILED DESCRIPTION

[0090] The anti-tumor effect produced by the combination of the antibody or fragment thereof against the CAPRIN-1 protein used in the present invention (hereinafter referred to as "anti-CAPRIN-1 antibody") and a RET kinase inhibitor is preferably evaluated by investigating the phagocytic activity of cancer cells produced by immune cells when cancer cells and immune cells are co-cultured in vitro, as described later. Among them, the immune cells used in the evaluation of the anti-tumor effect in vitro can be any cells as long as they are blood cells with phagocytic activity, preferably human monocytes (THP-1 or U937). If the antibody binds to the cancer cells, it is recognized by the immune cells, and the cancer cells are killed through the phagocytic activity produced by the immune cells, thereby evaluating the anti-tumor effect in vitro, and thus the anti-tumor effect in vivo can be predicted.

[0091] In this specification, "combination" or "combination" means that the anti-CAPRIN-1 antibody and the RET kinase inhibitor are administered or added to the same organism or cell at the same time or at a predetermined interval as independent active ingredients. The interval may be simultaneous administration, or 30 minutes later, or 1 hour later, 3 hours later, 6 hours later, 12 hours later, 1 day later, 2 days later, 3 days later, 5 days later, 7 days later, 2 weeks later, 3 weeks later, or 4 weeks later. When the anti-CAPRIN-1 antibody or the RET kinase inhibitor shows its anti-tumor effect, either one may be administered or added.

[0092] The term "comprising together or separately in combination" used in this specification means that a plurality of drugs are contained in a form that can be administered to a patient simultaneously or separately, and this form may be, for example, a so-called mixed preparation in which a plurality of drugs are mixed, or a so-called kit preparation (pharmaceutical kit) in which a plurality of drugs are contained as separate preparations. This form also includes a kit preparation in which a plurality of drugs are contained in any combination in two or more preparations.

[0093] Such a kit preparation involved in the present invention may be, for example, a kit preparation comprising: a preparation (or pharmaceutical composition) containing an anti-CAPRIN-1 antibody and a preparation (or pharmaceutical composition) containing a RET kinase inhibitor.

[0094] The anti-CAPRIN-1 antibody involved in the present invention can be a monoclonal antibody or a polyclonal antibody, preferably a monoclonal antibody. As long as the antibody of the present invention can exert an anti-tumor effect, it can be any type of antibody, and the antibody is a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, or a non-human animal antibody.

[0095] In addition, the subject to be treated and / or prevented for cancer in the present invention is a mammal such as primates, pets, livestock, and competitive animals, preferably a dog or a cat, and more preferably a human.

[0096] The following describes a pharmaceutical product comprising an anti-CAPRIN-1 antibody and a RET kinase inhibitor as active ingredients and a method for treating and / or preventing cancer according to the present invention.

[0097] <Anti-CAPRIN-1 Antibody>

[0098] As specific examples of antigens immunoreactive with the anti-CAPRIN-1 antibody used in the present invention, among the CAPRIN-1 proteins having the amino acid sequences represented by any of the even-numbered sequence numbers from SEQ ID NOs: 2 to 30, the amino acid sequences represented by SEQ ID NOs: 6, 8, 10, 12, and 14 are amino acid sequences of canine CAPRIN-1 proteins, the amino acid sequences represented by SEQ ID NOs: 2 and 4 are amino acid sequences of human CAPRIN-1 proteins, the amino acid sequence represented by SEQ ID NOs: 16 is the amino acid sequence of bovine CAPRIN-1 protein, the amino acid sequence represented by SEQ ID NOs: 18 is the amino acid sequence of horse CAPRIN-1 protein, the amino acid sequences represented by SEQ ID NOs: 20, 22, 24, 26, and 28 are the amino acid sequences of mouse CAPRIN-1 proteins, and the amino acid sequence represented by SEQ ID NOs: 30 is the amino acid sequence of chicken CAPRIN-1 protein.

[0099] In addition, the anti-CAPRIN-1 antibody used in the present invention may be an antibody that is immunoreactive with a variant of the CAPRIN-1 protein having a sequence identity of 80% or more, preferably 90% or more, more preferably 95% or more, and even more preferably 99% or more to the amino acid sequence represented by any of the even-numbered sequence numbers 2 to 30. Here, "% sequence identity" means the percentage (%) of identical amino acids (or bases) relative to the total number of amino acids (or bases) when two sequences are aligned (arranged) so as to maximize similarity (or identity) with or without introducing a gap.

[0100] The anti-CAPRIN-1 antibody in the present invention refers to an antibody or fragment thereof (antigen-binding fragment) that is immunoreactive with the full length or fragments of the CAPRIN-1 protein. Here, "immunoreactivity" refers to the property that the antibody specifically binds to the CAPRIN-1 protein or a partial polypeptide thereof in vivo.

[0101] The anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.

[0102] Polyclonal antibodies immunoreactive with the full length or fragments of the CAPRIN-1 protein (anti-CAPRIN-1 polyclonal antibodies) can be obtained by, for example, immunizing mice, mice producing human antibodies, rats, rabbits, chickens, etc. with natural CAPRIN-1 protein, or a fusion protein with GST or the like, or a partial peptide thereof, and then purifying the obtained serum by, for example, ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, affinity column coupled to the CAPRIN-1 protein or partial peptide, etc.

[0103] The base sequence and amino acid sequence of CAPRIN-1 and its homologues used in the above immunization can be obtained, for example, by registering in GenBank (NCBI, USA) and using algorithms such as BLAST and FASTA (Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90: 5873-5877, 1993; Altschul et al., Nucleic Acids Res. 25: 3389-3402, 1997). In addition, the method for producing the CAPRIN-1 protein can be obtained by referring to WO2014 / 012479, and cells expressing the CAPRIN-1 protein can also be used.

[0104] Monoclonal antibodies immunoreactive with the full length or fragments of CAPRIN-1 protein (anti-CAPRIN-1 monoclonal antibodies) can be obtained, for example, by administering breast cancer cells SK-BR-3 expressing CAPRIN-1, the full length or fragments of CAPRIN-1 protein, etc. to mice for immunization, and selecting clones producing anti-CAPRIN-1 monoclonal antibodies from the fusion cells (hybridomas) obtained by separating spleen cells from the mice and myeloma cells. Antibodies produced by the selected hybridomas can be obtained by the same method as the above-mentioned method for purifying polyclonal antibodies.

[0105] The antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies, non-human animal antibodies, and single-chain antibodies.

[0106] Human antibodies can be obtained by sensitizing human lymphocytes infected with Epstein-Barr virus with proteins, protein-expressing cells or their lysates, fusing the sensitized lymphocytes with myeloma cells such as human-derived U266 cells, and obtaining antibodies immunoreactive with the full-length CAPRIN-1 protein or fragments thereof from the resulting fused cells.

[0107] Humanized antibodies are also called reshaped human antibodies. Humanized antibodies are constructed by transplanting the complementary determining regions of antibodies derived from immune animals into the complementary determining regions of human antibodies. Genetic recombination methods as their general methods are also well-known technologies. Specifically, by PCR, several oligonucleotides are made in a way that the ends have overlapping parts, and a DNA sequence designed by connecting the complementary determining regions of, for example, mouse antibodies and rabbit antibodies to the framework regions of human antibodies is synthesized. The resulting DNA is connected to the DNA encoding the constant region of human antibodies, and then integrated into an expression vector, which is introduced into a host to produce it, thereby obtaining humanized antibodies (refer to European Patent Application Publication No. EP239400, International Publication No. WO96 / 02576). The framework region of human antibodies connected by the complementary determining regions can select the framework region that makes the complementary determining regions form a good antigen binding site. If necessary, amino acids in the framework region of the variable region of the antibody may be replaced so that the complementarity determining region of the reconstructed human antibody forms an appropriate antigen binding site (Sato K. et al., Cancer Research 1993, 53: 851-856). In addition, the framework region may be replaced with that derived from various human antibodies (see WO99 / 51743).

[0108] Antibodies are usually heteropolymeric glycoproteins containing at least 2 heavy chains and 2 light chains. Antibodies are composed of 2 identical light chains and 2 identical heavy chains. The heavy chain has a heavy chain variable region at one end, to which are attached several constant regions. The light chain has a light chain variable region at one end, to which are attached several constant regions. The variable region has specific variable regions called complementarity determining regions (CDRs) that confer binding specificity to the antibody. The relatively conserved portions of the variable region are called framework regions (FRs). The variable regions of complete heavy and light chains contain 4 FRs connected by 3 CDRs (CDR1 to CDR3) respectively.

[0109] In addition, the sequences of the constant regions and variable regions of the heavy and light chains of human origin can be obtained from, for example, NCBI (United States: GenBank, UniGene, etc.). For example, the heavy chain constant region of human IgG1 can refer to the sequence of accession number J00228, the heavy chain constant region of human IgG2 can refer to the sequence of accession number J00230, the human light chain κ constant region can refer to the sequences of accession numbers V00557, X64135, X64133, etc., and the human light chain λ constant region can refer to the sequences of accession numbers X64132, X64134, etc.

[0110] Chimeric antibodies are antibodies produced by combining sequences derived from different animals, for example, antibodies comprising the heavy chain variable region and light chain variable region of a mouse antibody and the heavy chain constant region and light chain constant region of a human antibody, etc. The production of chimeric antibodies can be carried out using known methods, for example, by connecting DNA encoding the antibody V region with DNA encoding the human antibody C region, integrating it into an expression vector and introducing it into a host, thereby producing it.

[0111] Non-human animal antibodies can be obtained by immunizing non-human animals with sensitizing antigens according to known methods. As a general method, sensitizing antigens can be injected into the peritoneal cavity, intradermal or subcutaneous of animals such as mice. When injecting sensitizing antigens, they can be mixed with various adjuvants represented by CFA (complete Freund's adjuvant) in appropriate amounts and administered to animals multiple times. After immunizing animals, serum is obtained after confirming that the serum contains anti-CAPRIN-1 antibodies. As mentioned above, it is purified by ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, affinity column coupled to CAPRIN-1 protein or partial peptide, etc., to obtain non-human animal antibodies. In addition, when obtaining monoclonal antibodies from non-human animals, immune cells can be collected from immunized animals and obtained by cell fusion with myeloma cells. The cell fusion of the immune cells and myeloma cells can be carried out according to known methods (refer to Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).

[0112] The antibodies used in the present invention can also be obtained as recombinant antibodies by cloning antibody genes from hybridomas, integrating them into appropriate vectors, introducing them into hosts, and producing them using gene recombination technology (see Carl, A.K. Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990).

[0113] The anti-CAPRIN-1 antibody used in the present invention may be an antibody in which amino acids in the variable region (e.g., FR) and the constant region are replaced with other amino acids. The amino acid replacement is 1 or more, for example, less than 15, less than 10, less than 8, less than 6, less than 5, less than 4, less than 3, or less than 2 amino acids, preferably 1 to 9 amino acids. The replaced antibody should be an antibody that has the same or higher binding affinity to the antigen as the unsubstituted antibody, and does not cause rejection when used in humans. The amino acid replacement is preferably a conservative amino acid replacement, which is a replacement between amino acids with similar properties such as charge, side chain, polarity, and aromaticity. Amino acids with similar properties can be classified into, 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 (leucine, isoleucine, alanine, valine, proline, phenylalanine, tryptophan, methionine), branched-chain amino acids (threonine, valine, isoleucine), aromatic amino acids (phenylalanine, tyrosine, tryptophan, histidine), etc.

[0114] The higher the binding affinity of the anti-CAPRIN-1 antibody used in the present invention to the CAPRIN-1 protein on the surface of cancer cells, the stronger the anti-tumor effect can be expected. The binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5×10 8 M -1 , at least 10 9 M -1 , at least 5×10 9 M -1 , at least 10 10 M -1 , at least 5×10 10 M -1 , at least 10 11 M -1 , at least 5×10 11 M -1 , at least 10 12 M -1 , or at least 10 13 M -1 .

[0115] The anti-CAPRIN-1 antibody used in the present invention may be chemically modified. Examples of such modified antibodies include antibodies bound to various molecules such as polyethylene glycol (PEG) and antitumor compounds (e.g., the antitumor agents exemplified below). In the modified antibody of the present invention, the substance to be bound is not limited. In order to obtain such modified antibodies, the obtained antibodies may be chemically modified. These methods have been established in the art.

[0116] The anti-CAPRIN-1 antibody used in the present invention can enhance the binding ability of the anti-CAPRIN-1 antibody to effector cells by replacing one, two or more amino acids in the heavy chain constant region of the antibody, or removing fucose bound to N-acetylglucosamine in the N-glycosidic bond type sugar chain bound to the heavy chain constant region. The above may be a single amino acid replacement, or a combination with an antibody bound to fucose.

[0117] Antibodies in which one, two or more amino acids in the heavy chain constant region are substituted can be prepared with reference to, for example, WO2004 / 063351, WO2011 / 120135, US Pat. No. 8,388,955, WO2011 / 005481, US Pat. No. 6,737,056, and WO2005 / 063351.

[0118] Antibodies from which fucose attached to N-acetylglucosamine in the N-glycosidic bond type sugar chain in the heavy chain constant region has been removed, or cells producing the antibodies, can be prepared with reference to U.S. Patent No. 6,602,684, European Patent No. 1,914,244, and U.S. Patent No. 7,579,170. Antibodies from which fucose bound to N-acetylglucosamine in the N-glycosidic bond type sugar chain bound to the heavy chain constant region has been removed, and compositions of antibodies to which fucose has been bound, or cells producing the compositions, can be prepared with reference to, for example, U.S. Patent No. 8,642,292.

[0119] The anti-CAPRIN-1 polyclonal antibody, anti-CAPRIN-1 monoclonal antibody, antibody preparation method, purification method, and preparation method of CAPRIN-1 protein or its partial polypeptide for immunization used in the present invention can refer to WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO 2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212.

[0120] Specific examples of the anti-CAPRIN-1 antibody in the present invention include the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212, but the following antibodies are mentioned as preferred anti-CAPRIN-1 antibodies.

[0121] An antibody or a fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as shown in SEQ ID NO: 2 or SEQ ID NO: 4, or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, even more preferably 95% or more, and even more preferably 99% or more) to the amino acid sequence.

[0122] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 31 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 36, 37, and 38, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 40, 41, and 42, or a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 140, 141, and 142, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3) of SEQ ID NOs: 143, 144, and 145. 45 (CDR1, CDR2 and CDR3, respectively), and an antibody or fragment thereof that is immunoreactive with the CAPRIN-1 protein; or an antibody or fragment thereof that contains a heavy chain variable region comprising the complementary determining regions of SEQ ID NOs: 164, 165 and 166 (CDR1, CDR2 and CDR3, respectively), and a light chain variable region comprising the complementary determining regions of SEQ ID NOs: 167, 168 and 169 (CDR1, CDR2 and CDR3, respectively), and is immunoreactive with the CAPRIN-1 protein. More preferably, an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 39 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 43; or an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 70 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 71; or an antibody or fragment thereof that contains a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 78 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 79.

[0123] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 33 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 60, 61, and 62, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 64, 65, and 66, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 63 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 67 in the light chain variable region.

[0124] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 32 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 52, 53, and 54, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 56, 57, and 58. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 55 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 59 in the light chain variable region.

[0125] An antibody or a fragment thereof immunoreactive with a partial polypeptide of the CAPRIN-1 protein having the amino acid sequence shown in SEQ ID NO: 34 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 170, 171 and 172 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 173, 174 and 175 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein; or a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 176, 177 and 178 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein. More preferably, it is an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 80 and the light chain variable region comprises the amino acid sequence of sequence number 81; or an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 82 and the light chain variable region comprises the amino acid sequence of sequence number 83.

[0126] An antibody or a fragment thereof immunoreactive with a partial polypeptide of the CAPRIN-1 protein having the amino acid sequence shown in SEQ ID NO: 35 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 182, 183 and 184 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 185, 186 and 187 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein; or a heavy chain variable region comprising the complementarity determining regions of SEQ ID NOs: 188, 189 and 190 (CDR1, CDR2 and CDR3, respectively) and a light chain variable region comprising the complementarity determining regions of SEQ ID NOs: 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively) and is immunoreactive with the CAPRIN-1 protein. More preferably, it is an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 84 and the light chain variable region comprises the amino acid sequence of sequence number 85; or an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of sequence number 86 and the light chain variable region comprises the amino acid sequence of sequence number 87.

[0127] An antibody or fragment thereof comprising a heavy chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 44, 45 and 46, and a light chain variable region comprising the complementarity determining regions (CDR1, CDR2 and CDR3, respectively) of SEQ ID NOs: 48, 49 and 50, and having immunoreactivity with the CAPRIN-1 protein. Preferably, the antibody or fragment thereof is one in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 47, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 51.

[0128] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 296 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 146, 147, and 148, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 149, 150, and 151. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 72 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 73 in the light chain variable region.

[0129] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 297 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 272, 273, and 274, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 275, 276, and 277. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 114 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 115 in the light chain variable region.

[0130] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 298 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 290, 291, and 292, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 293, 294, and 295, and immunoreactive with a CAPRIN-1 protein. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 120 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 121 in the light chain variable region.

[0131] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 299 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 301, 302, and 303, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 305, 306, and 307. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 300 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 304 in the light chain variable region.

[0132] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 308 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 134, 135, and 136, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 137, 138, and 139. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 68 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 69 in the light chain variable region.

[0133] An antibody or fragment thereof immunoreactive with a partial polypeptide of a CAPRIN-1 protein having an amino acid sequence as set forth in SEQ ID NO: 309 or an amino acid sequence having a sequence identity of 80% or more (preferably 85% or more, more preferably 90% or more, and even more preferably 95% or more) to the amino acid sequence. Preferably, the antibody or fragment thereof immunoreactive with a CAPRIN-1 protein comprises a heavy chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 134, 135, and 136, and a light chain variable region comprising the complementary determining regions (CDR1, CDR2, and CDR3, respectively) of SEQ ID NOs: 137, 138, and 139. More preferably, the antibody or fragment thereof comprises an amino acid sequence as set forth in SEQ ID NO: 68 in the heavy chain variable region and an amino acid sequence as set forth in SEQ ID NO: 69 in the light chain variable region.

[0134] In addition, the following anti-CAPRIN-1 antibodies are also preferably used.

[0135] 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.

[0136] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71.

[0137] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73.

[0138] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75.

[0139] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77.

[0140] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79.

[0141] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81.

[0142] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83.

[0143] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85.

[0144] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87.

[0145] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89.

[0146] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 91.

[0147] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 93.

[0148] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 95.

[0149] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 97.

[0150] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 99.

[0151] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101.

[0152] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103.

[0153] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105.

[0154] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107.

[0155] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109.

[0156] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111.

[0157] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113.

[0158] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115.

[0159] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117.

[0160] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119.

[0161] 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.

[0162] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123.

[0163] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125.

[0164] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127.

[0165] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129.

[0166] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131.

[0167] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133.

[0168] An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 304.

[0169] In the examples described below, the above-mentioned polyclonal antibodies or monoclonal antibodies directed against the full length of the CAPRIN-1 protein or a portion of the polypeptide expressed on the cell membrane surface of cancer cells were confirmed to show reactivity to the cell membrane surface of various human cancer cells. In addition, results showing efficacy were obtained in human cancer patients, and a significant anti-tumor effect was obtained in which the tumor completely disappeared in some cancer sites.

[0170] <RET kinase inhibitors>

[0171] The RET kinase inhibitor is a drug that has the effect of inhibiting the activity of RET (Rearranged During Transfection) kinase as described in detail below.

[0172] RET kinase is a receptor tyrosine kinase that binds to extracellular signal transduction molecules of the GDNF (glial cell line-derived neurotrophic factor) family and is encoded by the RET gene in humans. RET kinase is a transmembrane glycoprotein receptor composed of three main domains: an extracellular ligand binding domain, a transmembrane domain, and an intracellular tyrosine kinase domain. RET is a receptor for GDNF family ligands (GFL). In order to activate RET, GFL first needs to form a complex with a co-receptor fixed to the membrane by a GPI anchor. The co-receptor itself is classified into the GFRα (GDNF receptor-α) protein family. Various members of the GFRα family (GFRα1, GFRα2, GFRα3, GFRα4) each show specific binding activity to a specific GFL. Once the GFL-GFRα complex is formed, the complex binds two RET molecules and initiates reverse autophosphorylation of specific tyrosine residues in the tyrosine kinase domain of each RET molecule. The activated RET kinase activates signal transduction pathways such as the PI3K-Akt pathway and the MAPK pathway, controlling cell proliferation and the like.

[0173] It is known that RET kinase inhibitors inhibit signal transduction pathways by inhibiting dimer formation of RET kinase, neutralizing ligands, inhibiting ligand binding, or internalizing RET kinase, thereby exerting cancer treatment effects. However, in the present invention, the efficacy of cancer immunotherapy itself, specifically the phagocytic activity of cancer cells using immune cells as shown in the examples of the present application specification, can be enhanced. Since the signal transduction pathway activated by RET kinase has no direct correlation with the immune system, the enhancement of the efficacy of cancer immunotherapy itself produced by the present invention is a new cognition that cannot be anticipated by the current technical level of those skilled in the art.

[0174] Specific examples of RET kinase inhibitors include sorafenib (BAY 43-9006 / NSC-724772 / Nexavar), sunitinib (SU11248 / Sutent), vandetanib (ZD6474 / CH 331 / Caprelsa), regorafenib (BAY 73-4506 / Stivarga), lenvatinib (E7080 / ER-203492-00 / Kisplyx / Lenvima), selpercatinib (LOXO-292 / LOXO292 / LY3527723 / Retevmo / WO2018071447A1), and Compound Example 163), Agerafenib (RXDX-105), Amuvatinib (MP-470), Cabozantinib, Danusertib (PHA-739358), Enbezotinib, Erdafitinib (JNJ-42756493), Fedratinib (TG101348), Ilorasertib, Pralsetinib (BLU-667), trans-Pralsetinib (US20170121312A1, Compound Example 129), Regorafenib monohydrate ( Monohydrate), Vepafestinib (WO2019039439), Zeteletinib, Tafetinib (SIM-010603), Zeteletinibhemiadipate, AD80, AST-487 (NVP-AST487), BAW2881 (NVP-BAW2881), BBT594, CS-2660 (JNJ-38158471), GSK3179106, JNJ-38158471, ML786 dihydrochloride, ON123300, PF 477736, Pz-1, RET V804M-IN-1, RET-IN-1 (WO2018071447A1, Compound Example 552), RET-IN-10 (WO2021135938A1, compound18), RET-IN-11, RET-IN-12, RET-IN-13, RET-IN-14, RET-IN-15 (WO2021115457A1, compound 51), RET-IN-16, RET-IN-17 (WO2016038552A1, compound1), RET-IN-18 (WO2022017524A1, compound 1), RET-IN-3, RET-IN-4, RET-IN-5 (WO2021213476A1, compound 18), RET-IN-6 (CN113461670A, compound 321), RET-IN-7, RET-IN-8 (WO2021093720A1, compound I-1), RET-IN-9 (WO2021115457A1, compound 29), RPI-1, SPP-86, TG101209, TPX-0046, WF-47-JS03, WHI-P180 and their pharmaceutically acceptable (known) salts or (known) derivatives.

[0175] Among these RET kinase inhibitors, sorafenib, sunitinib, vandetanib, regorafenib, lenvatinib, selpatinib, agerafenib, amovatinib, cabozantinib, danusetinib, enbezotinib, erdafitinib, fitratinib, pralsetinib, ilolasetinib, vepafestinib, zeteletinib, tafitinib or pharmaceutically acceptable (known) salts or (known) derivatives thereof are preferred, sorafenib, sunitinib, vandetanib, regorafenib, lenvatinib, selpatinib or pharmaceutically acceptable (known) derivatives thereof are more preferred, and selpatinib or pharmaceutically acceptable (known) salts or (known) derivatives thereof are further preferred.

[0176] Selpercatinib (alias: LOXO-292, ARRY-192) selectively inhibits the activity of c-RET. The CAS number is 2152628-33-4, and the IUPAC name is 6-(2-hydroxy-2-methylpropoxy)-4-[6-[6-[(6-methoxypyridin-3-yl)methyl]-3,6-diazabicyclo[3.1.1]heptane-3-yl]pyridin-3-yl]pyrazolo[1,5-a]pyridine-3-carbonitrile. The molecular formula is C 29 H 31 N7O3, molecular weight is 525.6.

[0177] <Other medicines>

[0178] As the active ingredient of the drug of the present invention, in addition to the anti-CAPRIN-1 antibody and RET kinase inhibitor, it can also contain antitumor agents known in the literature, etc., within the range that does not hinder the effect of the drug of the present invention. Known antitumor agents are not particularly limited, and specific examples include 5-fluorouracil, irinotecan, oxaliplatin, carboplatin, cisplatin, nedaplatin, gemcitabine, paclitaxel, albumin paclitaxel, imiquimod, immune checkpoint inhibitors, doxorubicin, daunorubicin, cyclophosphamide, methotrexate, thiotepa, busulfan, improsulfan, piposulfan, benzodopa, carboquinone, meturedopa, uredopa, altretamine, triethylene melamine, triethylene phosphoramide, triethylene thiophosphoramide, trimethylolomelamine, blatacin, blatacinone, camptothecin, bryostatin, callystatin, cryptophycin1, cryptophycin8, dolastatin, duocarmycin, eleutherobin, pancratistatin, sarcodictyin, spongistatin, chlornaphamustine, chlornaphamustine zine), cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uramustine, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, calicheamicin, dynemycin icin), clodronic acid, esperamicin, aclarubicin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin, carmine, carzinophilin, chromomycin, dactinomycin, detorbicin, 6-diazo-5-oxo-L-norleucine, adriamycin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin C,Mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptozocin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, denopterin, pteropterin, trimetrex ate), fludarabine, 6-mercaptopurine, thiopurine, thioguanine, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calusterone, drostanolone propionate, cyclothiocarb, melastane, testolactone, aminoglutethimide, mitotane, trilostane, folinic acid acid), aceglucuronide, aldehyde phosphamide glycoside, aminolevulinic acid, eniluracil, amsacrine, bestrabucil, bisantrene, defofamine, demecolcine, diaziquone, elfornithine, elliptinium, epothilone, etoglucid, serrata polysaccharide, lonidamine, maytansine, ansamitocine, mitoguazone, mitoxantrone, mopidanmol, nitraerine, pentostatin, phenamet, pirarubicin, losoxantrone, podophyllinic acid acid), 2-ethylhydrazide, benzylhydrazine, razoxane, rhizoxin, sizolan, spirogermanium, tenuazonic acid, triaziquone, roridine A, anguidine, polyurethane, vindesine,Dacarbazine, mannomustine, dibromomannitol, dibromodulanol, pipobroman, gacytosine, docetaxel, gemcitabine, 6-thioguanine, mercaptopurine, vinblastine, etoposide, vincristine, vinorelbine, novantrone, teniposide, edatrexate, daunomycin, aminopterin, xeloda, ibandronate, difluoromethylornithine (DMFO), topoisomerase inhibitors, retinoic acid, folinic acid, and pharmaceutically acceptable (known) salts or (known) derivatives thereof.

[0179] <Antitumor Effect of the Present Invention>

[0180] The anti-tumor effect produced by the combination of the anti-CAPRIN-1 antibody of the present invention and the RET kinase inhibitor can be evaluated in vivo or in vitro. The anti-tumor effect in vivo can be evaluated by administering the anti-CAPRIN-1 antibody and the RET kinase inhibitor to an organism having cancer, measuring the size of the tumor after administration, and investigating the size of the cancer over time. In addition, the anti-tumor effect in vivo can also be evaluated by investigating the survival rate of the organism. In addition, it can also be evaluated by investigating the production capacity of cytokines or chemokines. Furthermore, it can also be evaluated by investigating the prevention of cancer, the prevention of metastasis, or the prevention of recurrence.

[0181] The anti-tumor effect in vitro can be evaluated by investigating the cytotoxic activity and phagocytic activity of immune cells to cancer cells when cancer cells are co-cultured with immune cells. Therefore, the anti-tumor effect produced by the combination of anti-CAPRIN-1 antibodies and RET kinase inhibitors can be evaluated by adding anti-CAPRIN-1 antibodies and RET kinase inhibitors by combination in the co-culture system of cancer cells and immune cells, investigating the cytotoxic activity or phagocytic activity of immune cells to cancer cells. The immune cells used here can be any cells as long as they are blood cells with cytotoxic activity and phagocytic activity, preferably human NK cells in the case of evaluating cytotoxic activity, and human monocytes (THP-1 or U937) in the case of evaluating phagocytic activity. If the antibody is combined with cancer cells, it is recognized by immune cells, and cancer cells are killed via the cytotoxic activity and phagocytic activity produced by immune cells, thus by evaluating the anti-tumor effect in vitro, so as to predict the anti-tumor effect in vivo.

[0182] The ability of the anti-CAPRIN-1 antibody used in the present invention to bind to CAPRIN-1 can be identified by a binding assay using, for example, ELISA, Western blotting, immunofluorescence, flow cytometry, and the like.

[0183] The combination of the anti-CAPRIN-1 antibody of the present invention and the RET kinase inhibitor has an enhanced in vitro antitumor effect compared to the anti-CAPRIN-1 antibody alone, and the enhancement rate is preferably 1.5 times or more, more preferably 2 times or more, and even more preferably 3 times or more.

[0184] <Medicines for the treatment and / or prevention of cancer>

[0185] The pharmaceutical of the present invention is intended for the treatment and / or prevention of cancer. The cancer that is the target of the pharmaceutical of the present invention is not particularly limited as long as it is a cancer (cell) that expresses the CAPRIN-1 protein, particularly a cancer (cell) that expresses the CAPRIN-1 protein on the cell membrane surface.

[0186] The term "treatment" as used herein refers to the treatment of cancer based on the aforementioned anti-tumor effect. In addition, the term "prevention" as used herein refers not only to the prevention of the occurrence of cancer but also to the prevention of the metastasis or recurrence of cancer.

[0187] As used herein, the terms "tumor" and "cancer" refer to malignant neoplasms and are used interchangeably.

[0188] As the cancer that becomes the object in the present invention, any cancer can be used as long as it is a cancer that expresses CAPRIN-1 protein on the cell membrane surface. Preferably, it is colorectal cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, kidney cancer, breast cancer, gastric cancer, bile duct cancer, thyroid cancer, melanoma, renal cell carcinoma, Hodgkin's lymphoma, head and neck cancer, mesothelioma, colon / rectal cancer, esophageal cancer, gastroesophageal junction cancer, hepatocellular carcinoma, glioblastoma, urothelial carcinoma, bladder cancer, uterine cancer, primary central nervous system lymphoma, primary testicular lymphoma, biliary tract cancer, brain tumor, leukemia, liver cancer, sarcoma, fibrosarcoma, mastocytoma, adrenocortical carcinoma, Ewing's sarcoma, testicular cancer, basal cell carcinoma, lymphoma, multiple myeloma, Paget's disease or skin cancer. In addition, the above-mentioned cancers can be primary cancer, metastatic cancer, metastatic cancer or recurrent cancer, postoperative cancer, or unresectable cancer. Furthermore, melanoma is often used synonymously with malignant melanoma or malignant melanoma.

[0189] The cancer more specifically includes, for example, cutaneous T / NK cell lymphoma, peripheral blood T cell lymphoma, multiple myeloma, Bowen's disease, squamous cell carcinoma, extramammary Paget's disease, mycosis fungoides, Sezary syndrome, T cell leukemia / lymphoma with skin lesions only, cutaneous B cell lymphoma (indolent group), cutaneous T cell lymphoma breast cancer, combined breast cancer, mixed malignant tumors of the breast, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, and neuroepithelial tumors. Glioblastoma, glioblastoma, neuroblastoma, ependymoma, neuronal tumor, fetal neuroectodermal tumor, schwannoma, neurofibroma, meningioma, chronic lymphocytic leukemia, lymphoma, digestive tract lymphoma, digestive organ lymphoma, small to medium cell lymphoma, cecal cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, sigmoid colon cancer, rectal cancer, ovarian epithelial cancer, germ cell tumor, stromal cell tumor, pancreatic duct cancer, invasive pancreatic duct cancer, pancreatic adenocarcinoma, acinar cell carcinoma, squamous cell carcinoma, giant cell tumor, intraductal pancreatic cancer Head mucinous tumor, mucinous cystadenocarcinoma, pancreatoblastoma, pancreatic head cell tumor, Frantz tumor, serous cystadenocarcinoma, solid papillary carcinoma, gastrinoma, glucagonoma, insulinoma, multiple endocrine neoplasia syndrome 1 (Wermer syndrome), non-functional islet cell tumor, somatostatinoma, VIP-producing tumor, cervical cancer, uterine body cancer, fibrosarcoma, bone and joint sarcoma, Ewing sarcoma, Wilms' tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia , chronic leukemia, spinal cord tumors, soft tissue malignant tumors, teratocarcinoma tumors, head and neck cancer including hypopharyngeal cancer, middle pharyngeal cancer, tongue cancer, upper pharyngeal cancer, oral cancer, lip cancer, sinus cancer, laryngeal cancer, renal pelvis ureter cancer, bladder cancer, urethral cancer, testicular tumor, malignant pleural mesothelioma, malignant bone tumor, uterine body cancer, childhood malignant solid tumors (rhabdomyosarcoma, neuroblastoma, hepatoblastoma, neuroblastoma, Wilms tumor, retinoblastoma, central nervous system embryonic cell tumor, Ewing sarcoma family tumor), etc., but not limited to these. In addition, it includes the above-mentioned cancer as the primary cancer that can be palpated, cancer present under the skin, cancer present in the skin, superficial cancer, cancer present in the dermis or cancer present in non-solid organs, and progressive cancer. In addition, it includes the above-mentioned cancer as the primary cancer that can be palpated through metastasis or recurrence, cancer present under the skin, cancer present in the skin, superficial cancer, cancer present in the dermis or cancer present in non-solid organs.

[0190] In addition, preferred subjects (patients) are mammals, such as primates, pets, livestock, and competitive animals, and humans, dogs, and cats are particularly preferred.

[0191] The drug of the present invention can be formulated by methods known to those skilled in the art. The drug of the present invention can be used parenterally in the form of a sterile solution or suspension injection in water or a pharmaceutically acceptable liquid other than water. In the drug of the present invention, each preparation or pharmaceutical composition can also be appropriately combined with its active ingredient (at least one of the anti-CAPRIN-1 antibody and RET kinase inhibitor) and, for example, a pharmacologically acceptable carrier, medium, or additive, specifically sterile water, physiological saline, isotonic solution, buffer (buffer, etc.), vegetable oil, oily liquid, antioxidant, solubilizer, emulsifier, suspending agent, surfactant, stabilizer, fragrance, excipient, binder, etc., preferably, it can be formulated by mixing them in a unit dosage method generally considered to be required for pharmaceutical implementation. The amount of active ingredient in these preparations is such an amount that an appropriate dosage within the indicated range can be obtained.

[0192] The sterile composition for injection can use a medium such as distilled water for injection, and is prescribed according to the usual preparation implementation. As the aqueous solution for injection, for example, physiological saline, isotonic solution containing glucose and / or other adjuvants, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride can be cited, and it can also be used in conjunction with suitable dissolution aids, such as alcohol, specifically ethanol, polyols such as propylene glycol, polyethylene glycol, nonionic surfactants such as polysorbate 80 (TM), HCO-60. As oily liquid, sesame oil and soybean oil can be cited, and it can also be used in conjunction with benzyl benzoate and benzyl alcohol as dissolution aids. In addition, it can also be mixed with buffers such as phosphate buffer, sodium acetate buffer, analgesics such as procaine hydrochloride, stabilizers such as benzyl alcohol, phenol, and antioxidants. The prepared injection is usually filled in a suitable ampoule.

[0193] Administration is oral or parenteral, preferably parenteral, and specifically, injection dosage forms, nasal administration dosage forms, pulmonary administration dosage forms, transdermal administration dosage forms, etc. can be cited. As an example of an injection dosage form, it can be administered systemically or locally by, for example, intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, intratumor injection, etc. As an example of a transdermal administration type, for example, it is a dosage form called a coating agent or an external medicine. As an external medicine, solid preparations, liquids, sprays, ointments, creams, or gels can be cited.

[0194] In addition, an appropriate administration method can be selected according to the patient's age, weight, sex, symptoms, etc. The administration amount of the pharmaceutical composition containing at least one of the anti-CAPRIN-1 antibody and RET kinase inhibitor is recorded in terms of the amount of each active ingredient, for example, it can be selected from the range of 0.0001 mg to 1000 mg per 1 kg of body weight at a time. Alternatively, for example, the administration amount of the active ingredient can be selected in the range of 0.001 to 100000 mg / individual per patient, or for example, 0.1 to 300 mg per 1 kg of body weight of the patient, or 1 mg to 30 mg, but it is not necessarily limited to these values. The administration amount and administration method vary according to the patient's weight, age, sex, symptoms, etc., and can be appropriately selected by those skilled in the art.

[0195] <How to give>

[0196] The treatment and / or prevention of cancer using the drug for the treatment and / or prevention of cancer of the present invention includes various forms in addition to the administration as the aforementioned drug. For example, the active ingredients of the drug of the present invention can be administered simultaneously or one by one in sequence. As a specific example, the second active ingredient can be administered within a time interval of about 3 weeks or less, that is, from the time just after the first active ingredient is administered to about 3 weeks. At this time, it can be implemented after the surgical treatment, or it can be implemented between the administration of the first agent and the second agent. In addition, the drug for the treatment and / or prevention of cancer of the present invention can be administered according to multiple administration cycles. For example, in the case of simultaneous administration of the active ingredients of the drug for the treatment and / or prevention of cancer of the present invention, the pharmaceutical composition containing the active ingredients of both parties is administered in a cycle of about 2 days to about 3 weeks. Then, according to the judgment of the responsible physician, the treatment cycle can be repeated as needed. Similarly, when planning a prescription in sequence, the administration period of each drug can be adjusted to the same period. The interval between cycles varies between 0 and 2 months. The dosage of each active ingredient in the pharmaceutical for treating and / or preventing cancer of the present invention can be set in the same manner as the dosage of each active ingredient in the above-mentioned pharmaceutical composition.

[0197] <Pharmaceutical Kit>

[0198] The drug for the treatment and / or prevention of cancer of the present invention may be in the form of a pharmaceutical kit. A pharmaceutical kit refers to a package for using the active ingredients in the form of a single pharmaceutical composition (preparation) in a method for treating and / or preventing cancer, and the package may include instructions for administering each active ingredient. Each active ingredient of the above-mentioned pharmaceutical composition for the treatment and / or prevention of cancer contained in the pharmaceutical kit may be in the form of a pharmaceutical composition formulated as described above in a manner that enables each active ingredient to be administered together or separately. In addition, the pharmaceutical kit contains an active ingredient in a sufficient amount for one or more doses in order to enable each active ingredient to be administered according to the above-mentioned administration method.

[0199] <Treatment and / or prevention methods>

[0200] Based on the contents specifically described above, the present invention provides a method for treating and / or preventing cancer comprising administering the above-mentioned pharmaceutical of the present invention, or the anti-CAPRIN-1 antibody of the present invention and a RET kinase inhibitor to a subject (patient). For example, the present invention further provides a method for treating and / or preventing cancer comprising administering the above-mentioned pharmaceutical of the present invention, etc. to a subject (patient) suspected of having cancer. In addition, in its embodiment, for example, the anti-CAPRIN-1 antibody (antibody or fragment thereof) of the present invention, the RET kinase inhibitor, and any antitumor agent contained in the above-mentioned pharmaceutical can be administered to the above-mentioned subject (patient) simultaneously or separately.

[0201] Example

[0202] Hereinafter, the present invention will be specifically described based on Examples, but the scope of the present invention is not limited to these specific examples.

[0203] (Example 1) Preparation of anti-CAPRIN-1 antibody

[0204] 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 Shigma Co.) and used as an antigen solution for each mouse. After the antigen solution was administered into the peritoneal cavity of 6-week-old Balb / c mice (manufactured by SLC Co., Ltd. of Japan), it was further administered 3 times and 24 times per week to complete the immunization. The spleens of each group removed 3 days after the last immunization were ground by clamping with two sterilized slides, and the operation of washing with PBS (-) (manufactured by Nissui Co., Ltd.), centrifuging at 1500 rpm for 10 minutes and removing the supernatant was repeated 3 times to obtain spleen cells. The obtained spleen cells were mixed with mouse myeloma cells SP2 / 0 (purchased from ATCC) at a ratio of 10:1, and a PEG solution prepared by mixing 200 μL of RPMI1640 medium containing 10% FBS and 800 μL of PEG1500 (made by Bellinga) warmed to 37°C was added thereto, and the mixture was allowed to stand for 5 minutes to perform cell fusion. Centrifugation was performed at 1700 rpm for 5 minutes, the supernatant was removed, and then the cells were suspended in 150 ml of RPMI1640 medium containing 15% FBS (HAT selection medium) to which 2% equivalent of HAT solution made by Gibco was added, and 100 μL of each well of a 96-well plate (made by Nunk) was inoculated in 15 plates. The hybridoma formed by the fusion of spleen cells and myeloma cells was obtained by culturing under the conditions of 7 days, 37°C, and 5% CO2. The hybridomas were screened using the binding affinity of the antibodies produced by the prepared hybridomas to the CAPRIN-1 protein as an index. 100 μL of a 1 μg / ml CAPRIN-1 protein solution was added to each well of a 96-well plate and allowed to stand at 4°C for 18 hours. After washing each well three times with PBS-T, 400 μL of a 0.5% bovine serum albumin (BSA) solution (manufactured by Shigma) was added to each well and allowed to stand at room temperature for 3 hours. The solution was removed, and each well was washed three times with 400 μL of PBS-T. Then, 100 μL of each culture supernatant of the hybridoma obtained above was added to each well and allowed to stand at room temperature for 2 hours. After washing each well three times with PBS-T, 100 μL of an HRP-labeled anti-mouse IgG (H+L) antibody (manufactured by Invitrogen) diluted 5000 times with PBS was added to each well and allowed to stand at room temperature for 1 hour. After washing the wells 3 times with PBS-T, 100 μl of TMB substrate solution (Thermo) was added to each well and allowed to stand for 15 to 30 minutes to perform a color reaction. After color development, 100 μl of 1N sulfuric acid was added to each well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorbance meter. As a result, multiple hybridomas that produced antibodies with high absorbance values ​​were screened out. The screened hybridomas were added to the plate at 0.5 per well of a 96-well plate for culture. After 1 week, hybridomas that formed single colonies in the wells were observed.The cells in these wells were further cultured, and hybridomas were screened using the binding affinity of the antibodies produced by the cloned hybridomas to the CAPRIN-1 protein as an index. 100 μL of a 1 μg / ml CAPRIN-1 protein solution was added to each well of a 96-well plate and allowed to stand at 4°C for 18 hours. After washing each well three times with PBS-T, 400 μL of a 0.5% BSA solution was added to each well and allowed to stand at room temperature for 3 hours. The solution was removed, and each well was washed three times with 400 μL of PBS-T, and 100 μL of each culture supernatant of the hybridoma obtained above was added to each well and allowed to stand at room temperature for 2 hours. After washing each well three times with PBS-T, 100 μL of an HRP-labeled anti-mouse IgG (H+L) antibody (manufactured by Invitrogen) diluted 5000 times with PBS was added to each well and allowed to stand at room temperature for 1 hour. After washing the wells three times with PBS-T, 100 μL of TMB substrate solution (manufactured by Thermo) was added to each well and allowed to stand for 15 to 30 minutes to perform a color reaction. After color development, 100 μL of 1N sulfuric acid was added to each well to stop the reaction, and the absorbance values ​​at 450 nm and 595 nm were measured using an absorbance meter. As a result, a plurality of mouse monoclonal antibodies reactive with the CAPRIN-1 protein were obtained.

[0205] Furthermore, the reactivity to human cancer cells, which are cells confirmed to express CAPRIN-1 protein on the cell membrane surface, was confirmed by flow cytometry. A mouse IgG control antibody that does not show reactivity with the cancer cells was used as a negative control. As a result of the confirmation, several monoclonal antibodies were obtained that had a stronger fluorescence intensity than the mouse IgG control antibody and strongly reacted with the cell membrane surface of the above cancer cells expressing CAPRIN-1 on the cell membrane surface. From these, as monoclonal antibodies that show reactivity with CAPRIN-1 protein, antibodies containing the amino acid sequence of the heavy chain variable region shown in sequence number 114 and the amino acid sequence of the light chain variable region shown in sequence number 115, which are monoclonal antibodies against CAPRIN-1 described in WO2013 / 125630, were screened.

[0206] The CDR1 to 3 of the heavy chain variable region of the above-screened antibody was identified, and the base sequence was designed in a manner that the heavy chain variable region containing the sequence of a human antibody in the framework region could be expressed, and it was inserted into a mammalian expression vector into which the heavy chain constant region of human IgG1 had been inserted. Similarly, the CDR1 to 3 of the light chain variable region was identified, and the base sequence was designed in a manner that the light chain variable region containing the sequence of a human antibody in the framework region could be expressed, and it was inserted into a mammalian expression vector into which the light chain constant region of human IgG1 had been inserted. The above two recombinant expression vectors were introduced into mammalian cells according to conventional methods to obtain a culture supernatant containing humanized monoclonal antibody #1 (humanized antibody #1) against CAPRIN-1. The amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain of humanized monoclonal antibody #1 are shown in sequence numbers 272, 273, and 274, respectively. The amino acid sequences of CDR1, CDR2, and CDR3 of the light chain of humanized monoclonal antibody #1 are shown in SEQ ID NOs: 275, 276, and 277, respectively.

[0207] The obtained culture supernatant containing humanized anti-CAPRIN-1 monoclonal antibody #1 was purified using Hitrap Protein A Sepharose FF (GE Healthcare) according to a conventional method, replaced with PBS (-), and filtered through a 0.22 μm filter (Millipore).

[0208] The specific reactivity of the anti-CAPRIN-1 antibody to the CAPRIN-1 protein was confirmed by immobilizing the CAPRIN-1 protein on a plate and detecting it using the ELISA method.

[0209] Furthermore, the anti-CAPRIN-1 antibody was used to examine its reactivity with cancer cells that had not been treated to permeabilize the cell membrane by flow cytometry, thereby confirming that a portion of the CAPRIN-1 protein was expressed on the cell membrane surface of cancer cells as shown in the following Examples.

[0210] The results of flow cytometry showed that the expression of CAPRIN-1 gene in breast cancer cells (BT-474), colorectal cancer cells (HT-29, HCT116), lung cancer cells (QG56, H1650, A549), gastric cancer cells (NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1, DU145), 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), and esophageal cancer cells (E-10). Humanized antibody #1 showed stronger fluorescence intensity against any of the cancer cells, including ductal carcinoma cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder carcinoma cells (TGBC14TKB), fibrosarcoma cells (HT-1080), melanoma cells (G-361, A375), mouse renal carcinoma cells (Renca) in which the expression of the CAPRIN-1 gene was confirmed, and mouse breast carcinoma cells (4T1), than the human IgG control antibody and rabbit IgG antibody that showed no reactivity against the cancer cells as negative controls, and it was confirmed that it strongly reacted with the cell membrane surface of the above cancer cells expressing CAPRIN-1.

[0211] In addition, for WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 0 The anti-CAPRIN-1 antibodies described in 18891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176, and WO2015 / 020212 were also confirmed to react strongly with the cancer cell membrane surface.

[0212] (Example 2A) In vitro anti-tumor effect of anti-CAPRIN-1 antibody combined with serpatinib

[0213] The anti-tumor effect of the combination of an anti-CAPRIN-1 antibody and selpatinib, a RET kinase inhibitor, was evaluated in vitro. Serpatinib purchased from Celec (Cat. No.: S8781) was used.

[0214] Specifically, in the RET kinase inhibitor combination test, human cancer cells that had been previously treated with each combination drug were co-cultured with human monocytes (THP-1) in the presence of anti-CAPRIN-1 antibodies, and the phagocytic activity of cancer cells by THP-1 via the antibody was evaluated.

[0215] As human-derived cancer cells, HCT116, a colon cancer cell line, was used. Human-derived cancer cells were cultured in the presence of serpatinib (5 μM) for 2 days in a selpatinib combination test area on a 6-well plate. In addition, a combination drug non-addition test area in which only anti-CAPRIN-1 antibodies were added was set as a control, and cancer cells were cultured for 2 days without adding any combination drugs.

[0216] The cultured cancer cell lines were stripped with "TrypLE Express" (Thermo), and Calcein-AM was added at a final concentration of 0.04 μg / mL and incubated at 37°C for 10 minutes to stain the cancer cells. 3 Cancer cells were dispensed into a 96-well plate, and anti-CAPRIN-1 antibody and 1.25×10 5 THP-1 cells were cultured at 37°C and 5% CO2 for 1 hour. Then, the cells were washed with PBS (phosphate buffer) containing 1% FBS (fetal bovine serum), and the human monocytes were stained by reacting with APC (allophycocyanin) labeled anti-human CD45 antibody at a final concentration of 0.25μg / mL. After further staining the dead cells with propidium iodide (PI) at a final concentration of 0.1μg / mL, the fluorescence of each cell was measured by flow cytometry. PI-positive dead cells were excluded during the analysis. THP-1, which is a human monocyte, recognizes the antibody bound to the cancer cell and phagocytoses the cancer cell. At this time, if the cancer cell is stained with Calcein-AM, the monocytes that have taken it up by phagocytosis also become Calcein-AM positive. Therefore, the phagocytic activity in this evaluation system is calculated by the ratio (%) of APC positive / Calcein-AM positive in all Calcein-AM positive groups.

[0217] The results of the evaluation using the anti-CAPRIN-1 antibody prepared in Example 1 (anti-CAPRIN-1 humanized antibody #1) showed that the phagocytic activity of cancer cells in HCT116 was less than 39% in the non-addition test area of ​​the combination drug, while the phagocytic activity of cancer cells in the serpatinib combination test area was more than 76%. The results of the Student's t-test showed that the phagocytic activity in the serpatinib combination test area was significantly higher than that in the non-addition test area of ​​the combination drug (p < 0.001; significance level 5%) ( Figure 1 ). Therefore, the anti-tumor effect of the anti-CAPRIN-1 antibody on the above-mentioned human cancer cells was significantly enhanced by about 1.9 times by the combination with selpatinib.

[0218] In addition, in WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534 , WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO In the combination of the anti-CAPRIN-1 antibodies described in 2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212 and selpatinib, the same enhancement of cancer cell phagocytic activity as in the combination of the anti-CAPRIN-1 humanized antibody #1 and selpatinib against the above-mentioned human cancer cells was observed.

[0219] (Example 2B) In vitro anti-tumor effect of anti-CAPRIN-1 antibody combined with serpatinib

[0220] The anti-tumor effect of the combination of an anti-CAPRIN-1 antibody and selpatinib, a RET kinase inhibitor, was evaluated in vitro. Serpatinib purchased from Celec (Cat. No.: S8781) was used.

[0221] Specifically, in the RET kinase inhibitor combination test, human cancer cells that had been previously treated with each combination drug were co-cultured with human monocytes (THP-1) in the presence of anti-CAPRIN-1 antibodies, and the phagocytic activity of cancer cells by THP-1 via the antibody was evaluated. Human IgG antibody (Shigma Cat. No.: 4506) was used as a control antibody.

[0222] As human-derived cancer cells, HCT116, a colon cancer cell line, was used. Human-derived cancer cells were cultured in the presence of serpatinib (5 μM) for 2 days in a selpatinib combination test area on a 6-well plate. In addition, a combination drug non-addition test area in which only anti-CAPRIN-1 antibodies were added was set as a control, and cancer cells were cultured for 2 days without adding any combination drugs.

[0223] The cultured cancer cell lines were stripped with "TrypLE Express" (Thermo), and Calcein-AM was added at a final concentration of 0.04 μg / mL and incubated at 37°C for 10 minutes to stain the cancer cells. 3 Cancer cells were dispensed into a 96-well plate, and anti-CAPRIN-1 antibody (or control IgG antibody at the same concentration) at a final concentration of 1 μg / mL and 1.25×10 5 THP-1 cells were cultured at 37°C and 5% CO2 for 1 hour. Then, the cells were washed with PBS (phosphate buffer) containing 1% FBS (fetal bovine serum), and the human monocytes were stained by reacting with APC (allophycocyanin) labeled anti-human CD45 antibody at a final concentration of 0.25μg / mL. After further staining the dead cells with propidium iodide (PI) at a final concentration of 0.1μg / mL, the fluorescence of each cell was measured by flow cytometry. PI-positive dead cells were excluded during the analysis. THP-1, which is a human monocyte, recognizes the antibody bound to the cancer cell and phagocytoses the cancer cell. At this time, if the cancer cell is stained with Calcein-AM, the monocytes that have taken it up by phagocytosis also become Calcein-AM positive. Therefore, the phagocytic activity in this evaluation system is calculated by the ratio (%) of APC positive / Calcein-AM positive in all Calcein-AM positive groups.

[0224] As a result of the evaluation using the anti-CAPRIN-1 antibody prepared in Example 1 (anti-CAPRIN-1 humanized antibody #1), the phagocytic activity of cancer cells in HCT116 was observed to be less than 39% in the test area without adding the combination drug, while the phagocytic activity of cancer cells in the test area with serpatinib was observed to be more than 76%. The Dunnett's test showed that the phagocytic activity in the test area with serpatinib was significantly higher than that in the test area without adding the combination drug (p < 0.001; significance level 5%) ( Figure 2 ). Therefore, the anti-tumor effect of the anti-CAPRIN-1 antibody on the above-mentioned human cancer cells was significantly enhanced by about 1.9 times by the combination with selpatinib.

[0225] (Example 3) In vitro antitumor effects of antibodies alone or in combination with serpatinib

[0226] The anti-tumor effects of anti-CAPRIN-1 antibody, anti-CD20 antibody and anti-HER-2 antibody alone or in combination with selpatinib were evaluated in vitro. Serpatinib was purchased from Celec (Cat. No.: S8781). Anti-CD20 antibody ("rituximab") and anti-HER-2 antibody ("Herceptin") were purchased from Chugai Pharmaceutical Co., Ltd.

[0227] Specifically, in the selpatinib combination test, human cancer cells that had been previously treated with selpatinib were co-cultured with human monocytes (THP-1) in the presence of 1 μM anti-CAPRIN-1 antibody, anti-CD20 antibody, or anti-HER-2 antibody, and the phagocytic activity of cancer cells produced by THP-1 via the antibody was evaluated.

[0228] As human-derived cancer cells, HCT116, a colon cancer cell line, was used. On a 6-well plate, human-derived cancer cells were cultured for 2 days in the presence of combined serpatinib in the serpatinib combined test area. Specifically, serpatinib (5 μM) was added to HCT116. In addition, as a control, a non-added test area for the combined drug in which only anti-CAPRIN-1 antibody, anti-CD20 antibody, or anti-HER-2 antibody was added was set up, and cancer cells were cultured for 2 days without adding serpatinib in advance.

[0229] The cultured cancer cell lines were stripped with "TrypLE Express" (Thermo), and Calcein-AM was added at a final concentration of 0.04 μg / mL and incubated at 37°C for 10 minutes to stain the cancer cells. 4 Cancer cells were dispensed into a 96-well plate, and anti-CAPRIN-1 antibody, anti-CD20 antibody, or anti-HER-2 antibody at a final concentration of 1 μg / mL and 1×10 5THP-1 cells were cultured at 37°C and 5% CO2 for 1 hour. Then, the cells were washed with PBS (phosphate buffer) containing 1% FBS (fetal bovine serum), and the human monocytes were stained by reacting with APC (allophycocyanin) labeled anti-human CD45 antibody at a final concentration of 0.25μg / mL. After further staining the dead cells with propidium iodide (PI) at a final concentration of 0.1μg / mL, the fluorescence of each cell was measured by flow cytometry. PI-positive dead cells were excluded during the analysis. THP-1, which is a human monocyte, recognizes the antibody bound to the cancer cell and phagocytoses the cancer cell. At this time, if the cancer cell is stained with Calcein-AM, the monocytes that have taken it up by phagocytosis also become Calcein-AM positive. Therefore, the phagocytic activity in this evaluation system is calculated by the ratio (%) of APC positive / Calcein-AM positive in all Calcein-AM positive groups.

[0230] As a result of the evaluation using the anti-CAPRIN-1 antibody prepared in Example 1 (anti-CAPRIN-1 humanized antibody #1), less than 21% of cancer cell phagocytosis activity was observed in the HCT116 in the non-addition test area of ​​the combination drug, whereas more than 50% of cancer cell phagocytosis activity was observed in the serpatinib combination test area. As a result of the Dunnett's test, the phagocytosis activity in the serpatinib combination test area was significantly higher than that in the non-addition test area of ​​the combination drug (all p < 0.001; significance level 5%) ( Figure 3 ).

[0231] On the other hand, the results of the evaluation using anti-CD20 antibodies or anti-HER-2 antibodies showed that, for anti-CD20 antibodies, less than 7% of cancer cell phagocytosis activity was observed in HCT116 in the non-addition test area of ​​the combination drug, while less than 18% of cancer cell phagocytosis activity was observed in the selpatinib combination test area, and no significant enhancement of the drug efficacy due to the combination was observed. For anti-HER-2 antibodies, less than 10% of cancer cell phagocytosis activity was observed in HCT116 in the non-addition test area of ​​the combination drug, while less than 15% of cancer cell phagocytosis activity was observed in the selpatinib combination test area, and no significant enhancement of the drug efficacy due to the combination was observed ( Figure 3 ).

[0232] The above results suggest that the enhancement of the anti-tumor effect produced by the combination with selpatinib is an effect specific to the anti-CAPRIN-1 antibody.

[0233] In addition, in WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, In the combination of the anti-CAPRIN-1 antibodies described in WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212 and selpatinib, enhancement of phagocytic activity similar to the combination of the anti-CAPRIN-1 humanized antibody #1 and selpatinib against the above-mentioned human cancer cells was also observed.

[0234] (Example 4) In vitro antitumor effect of the combination of anti-CAPRIN-1 antibody and selpatinib or cisplatin

[0235] The anti-tumor effect of the combination of anti-CAPRIN-1 antibody and selpatinib or cisplatin was evaluated in vitro. Serpatinib was purchased from Celec (Cat. No.: S8781) and cisplatin was purchased from Physica.

[0236] Specifically, in each inhibitor combination test area, human cancer cells previously treated with each combination drug were co-cultured with human monocytes (THP-1) in the presence of anti-CAPRIN-1 antibodies, and the phagocytic activity of cancer cells by THP-1 via the antibody was evaluated.

[0237] As human-derived cancer cells, HCT116, a colon cancer cell line, was used. Human-derived cancer cells were cultured for 2 days in the presence of various combination drugs in various drug combination test areas on a 6-well plate. Specifically, selpatinib was added to HCT116 at a concentration of 5 μM and cisplatin was added at a concentration of 1 μM. In addition, a combination drug non-addition test area was set up as a control, in which only anti-CAPRIN-1 antibodies were added, and cancer cells were cultured for 2 days without adding each combination drug.

[0238] The cultured cancer cell lines were stripped with "TrypLE Express" (Thermo), and Calcein-AM was added at a final concentration of 0.04 μg / mL and incubated at 37°C for 10 minutes to stain the cancer cells. 4Cancer cells were dispensed into a 96-well plate, and anti-CAPRIN-1 antibody and 1×10 5 THP-1 cells were cultured at 37°C and 5% CO2 for 1 hour. Then, the cells were washed with PBS (phosphate buffer) containing 1% FBS (fetal bovine serum), and the human monocytes were stained by reacting with APC (allophycocyanin) labeled anti-human CD45 antibody at a final concentration of 0.25μg / mL. After further staining the dead cells with propidium iodide (PI) at a final concentration of 0.1μg / mL, the fluorescence of each cell was measured by flow cytometry. PI-positive dead cells were excluded during the analysis. THP-1, which is a human monocyte, recognizes the antibody bound to the cancer cell and phagocytoses the cancer cell. At this time, if the cancer cell is stained with Calcein-AM, the monocytes that have taken it up by phagocytosis also become Calcein-AM positive. Therefore, the phagocytic activity in this evaluation system is calculated by the ratio (%) of APC positive / Calcein-AM positive in all Calcein-AM positive groups.

[0239] The result of evaluating the anti-CAPRIN-1 antibody (anti-CAPRIN-1 humanized antibody #1) prepared in Example 1 as an anti-CAPRIN-1 antibody was that less than 21% of cancer cell phagocytic activity was observed in HCT116 in the combination drug non-addition test area, while more than 50% of cancer cell phagocytic activity was observed in the selpatinib combination test area. On the other hand, less than 23% of cancer cell phagocytic activity was observed in the cisplatin combination test area. The result of Dunnett's test was that the phagocytic activity in the selpatinib combination test area was significantly higher than that in the combination drug non-addition test area (p < 0.001; significance level 5%), while no significant change was found in the phagocytic activity in the cisplatin combination test area compared with the combination drug non-addition test area ( Figure 4 ).

[0240] The above results suggest that the enhancement of the anti-tumor effect produced by the combination of drugs is a RET kinase inhibitor-specific effect.

[0241] In addition, in WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, The anti-CAPRIN-1 antibodies described in WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125640, WO2013 / 147169, WO2013 / 147176 and WO2015 / 020212 were also observed to show changes in phagocytic activity similar to the combination of the anti-CAPRIN-1 humanized antibody #1 against human cancer cells and various drugs.

[0242] All journals, patents and patent applications cited in this specification are incorporated herein by reference.

Claims

1. A pharmaceutical for treating and / or preventing cancer, comprising, together or separately, an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein, and a RET (Rearranged During Transfection) kinase inhibitor.

2. The drug according to claim 1, wherein the RET kinase inhibitor is Selpercatinib.

3. The pharmaceutical according to claim 1 or 2, wherein the antibody or fragment thereof is immunoreactive with a CAPRIN-1 protein having an amino acid sequence represented by any of even-numbered sequence numbers 2 to 30, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence. 4 . The pharmaceutical according to claim 1 , wherein the antibody or fragment thereof is immunoreactive with the extracellular region of the CAPRIN-1 protein present on the surface of cancer cells.

5. The pharmaceutical according to any one of claims 1 to 4, wherein the antibody or fragment thereof is immunoreactive with a partial polypeptide of the CAPRIN-1 protein having an amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, and 309, or an amino acid sequence having a sequence identity of 80% or more to the amino acid sequence. 6 . The pharmaceutical according to claim 1 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.

7. The pharmaceutical according to any one of claims 1 to 6, wherein the antibody or fragment thereof is any one of the following (A) to (M), (A) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 36, a complementarity determining region CDR2 set forth in SEQ ID NO: 37, and a complementarity determining region CDR3 set forth in SEQ ID NO: 38, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 40, a complementarity determining region CDR2 set forth in SEQ ID NO: 41, and a complementarity determining region CDR3 set forth in SEQ ID NO: 42; (B) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 44, a complementarity determining region CDR2 set forth in SEQ ID NO: 45, and a complementarity determining region CDR3 set forth in SEQ ID NO: 46, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 48, a complementarity determining region CDR2 set forth in SEQ ID NO: 49, and a complementarity determining region CDR3 set forth in SEQ ID NO: 50; (C) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 52, a complementarity determining region CDR2 set forth in SEQ ID NO: 53, and a complementarity determining region CDR3 set forth in SEQ ID NO: 54, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 56, a complementarity determining region CDR2 set forth in SEQ ID NO: 57, and a complementarity determining region CDR3 set forth in SEQ ID NO: 58; (D) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 60, a complementarity determining region CDR2 set forth in SEQ ID NO: 61, and a complementarity determining region CDR3 set forth in SEQ ID NO: 62, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 64, a complementarity determining region CDR2 set forth in SEQ ID NO: 65, and a complementarity determining region CDR3 set forth in SEQ ID NO: 66; (E) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 170, a complementarity determining region CDR2 set forth in SEQ ID NO: 171, and a complementarity determining region CDR3 set forth in SEQ ID NO: 172, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 173, a complementarity determining region CDR2 set forth in SEQ ID NO: 174, and a complementarity determining region CDR3 set forth in SEQ ID NO: 175; (F) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 176, a complementarity determining region CDR2 set forth in SEQ ID NO: 177, and a complementarity determining region CDR3 set forth in SEQ ID NO: 178, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 179, a complementarity determining region CDR2 set forth in SEQ ID NO: 180, and a complementarity determining region CDR3 set forth in SEQ ID NO: 181; (G) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 182, a complementarity determining region CDR2 set forth in SEQ ID NO: 183, and a complementarity determining region CDR3 set forth in SEQ ID NO: 184, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 185, a complementarity determining region CDR2 set forth in SEQ ID NO: 186, and a complementarity determining region CDR3 set forth in SEQ ID NO: 187; (H) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 188, a complementarity determining region CDR2 set forth in SEQ ID NO: 189, and a complementarity determining region CDR3 set forth in SEQ ID NO: 190, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 191, a complementarity determining region CDR2 set forth in SEQ ID NO: 192, and a complementarity determining region CDR3 set forth in SEQ ID NO: 193; (I) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 146, a complementarity determining region CDR2 set forth in SEQ ID NO: 147, and a complementarity determining region CDR3 set forth in SEQ ID NO: 148, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 149, a complementarity determining region CDR2 set forth in SEQ ID NO: 150, and a complementarity determining region CDR3 set forth in SEQ ID NO: 151; (J) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 272, a complementarity determining region CDR2 set forth in SEQ ID NO: 273, and a complementarity determining region CDR3 set forth in SEQ ID NO: 274, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 275, a complementarity determining region CDR2 set forth in SEQ ID NO: 276, and a complementarity determining region CDR3 set forth in SEQ ID NO: 277; (K) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 290, a complementarity determining region CDR2 set forth in SEQ ID NO: 291, and a complementarity determining region CDR3 set forth in SEQ ID NO: 292, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 293, a complementarity determining region CDR2 set forth in SEQ ID NO: 294, and a complementarity determining region CDR3 set forth in SEQ ID NO: 295; (L) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 301, a complementarity determining region CDR2 set forth in SEQ ID NO: 302, and a complementarity determining region CDR3 set forth in SEQ ID NO: 303, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 305, a complementarity determining region CDR2 set forth in SEQ ID NO: 306, and a complementarity determining region CDR3 set forth in SEQ ID NO: 307; (M) an antibody or a fragment thereof that is immunoreactive with a CAPRIN-1 protein and comprises a heavy chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 134, a complementarity determining region CDR2 set forth in SEQ ID NO: 135, and a complementarity determining region CDR3 set forth in SEQ ID NO: 136, and a light chain variable region comprising a complementarity determining region CDR1 set forth in SEQ ID NO: 137, a complementarity determining region CDR2 set forth in SEQ ID NO: 138, and a complementarity determining region CDR3 set forth in SEQ ID NO:

139.

8. The pharmaceutical according to any one of claims 1 to 7, wherein the antibody or fragment thereof is any one of the following (a) to (a1), (a) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 39 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43; (b) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:47 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:51; (c) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; (d) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 63 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 67; (e) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69; (f) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 70 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 71; (g) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 72 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 73; (h) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 74 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75; (i) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 77; (j) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 78 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 79; (k) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 80 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 81; (l) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 82 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 83; (m) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 84 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 85; (n) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 86 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 87; (o) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 89; (p) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 91; (q) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:92 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:93; (r) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:94 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:95; (s) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:96 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:97; (t) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 98 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 99; (u) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 101; (v) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 102 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 103; (w) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 104 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105; (x) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 106 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 107; (y) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 109; (z) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 110 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 111; (aa) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 112 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 113; (ab) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115; (ac) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 117; (ad) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 118 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 119; (ae) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 120 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 121; (af) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 122 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 123; (ag) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 124 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 125; (ah) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 127; (ai) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 129; (aj) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 130 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 131; (ak) an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 132 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133; (a1) An antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 300 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:

304. 9 . The pharmaceutical according to claim 1 , wherein the antibody is a human antibody, a humanized antibody, a chimeric antibody or a single-chain antibody. 10 . The pharmaceutical according to claim 1 , wherein the cancer is a cancer expressing CAPRIN-1 protein on the cell membrane surface.

11. The drug according to any one of claims 1 to 10, wherein the cancer is colorectal cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, kidney cancer, breast cancer, gastric cancer, bile duct cancer, thyroid cancer, melanoma, renal cell carcinoma, Hodgkin's lymphoma, head and neck cancer, mesothelioma, colon / rectal cancer, esophageal cancer, esophagogastric junction cancer, hepatocellular carcinoma, glioblastoma, urothelial carcinoma, bladder cancer, uterine cancer, primary central nervous system lymphoma, primary testicular lymphoma, biliary tract cancer, brain tumor, leukemia, liver cancer, sarcoma, fibrosarcoma, mast cell tumor, adrenocortical carcinoma, Ewing's sarcoma, testicular cancer, basal cell carcinoma, lymphoma, multiple myeloma, Paget's disease or skin cancer.

12. A drug efficacy enhancer for a pharmaceutical composition for treating and / or preventing cancer, the pharmaceutical composition for treating and / or preventing cancer comprising an antibody or a fragment thereof having immunoreactivity with a CAPRIN-1 protein as an active ingredient, wherein the drug efficacy enhancer comprises a RET kinase inhibitor as an active ingredient.

13. A drug efficacy enhancer for a pharmaceutical composition for treating and / or preventing cancer, wherein the pharmaceutical composition for treating and / or preventing cancer comprises a RET kinase inhibitor as an active ingredient, and wherein the drug efficacy enhancer comprises an antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein as an active ingredient.

14. A method for treating and / or preventing cancer, characterized in that: An antibody or a fragment thereof immunoreactive with a CAPRIN-1 protein and a RET kinase inhibitor are administered to a subject together or separately.

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