Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
  • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
  • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
  • A61K2039/507—Comprising a combination of two or more separate antibodies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
  • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
  • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Definitions

• the present invention relates to an antibody against the CAPRIN-1 protein or a fragment thereof, and a drug for treating and / or preventing cancer using an angiogenesis inhibitor and a taxane-based drug.
• Cytoplasmic-activation and proliferation-associated patent 1 (CAPRIN-1) is expressed on the cell membrane surface of many solid cancers, and an antibody against this CAPRIN-1 protein is used for cancer treatment and / or preventive medicine. It is known to be promising (Patent Document 1).
• a treatment method in which a plurality of cancer therapeutic agents are used in combination is used as a standard treatment method.
• a combination of irinotecan, phoric acid, and fluorouracil for breast cancer, a combination of doxorubicin and cyclophosphamide, or a combination of paclitaxel, trastuzumab, and veltuzumab.
• Gastric cancer is generally treated with a plurality of anticancer agents such as carboplatin and fluorouracil.
• a cancer therapeutic agent containing an anti-CAPRIN-1 antibody as an active ingredient also has an excellent cancer therapeutic effect when used in combination with a chemotherapeutic agent (Patent Document 2).
• Patent Document 2 the treatment of cancer with a combination of chemotherapeutic agents is not effective for all applicable cancers, and although it may enhance the therapeutic effect additively, it is synergistically significant. There are few things that enhance the therapeutic effect.
• One specific example of a cancer treatment method in which a plurality of cancer therapeutic agents are used in combination is a combination of an angiogenesis inhibitor (for example, ramucirumab) and a taxane drug (for example, paclitaxel).
• angiogenesis inhibitor for example, ramucirumab
• a taxane drug for example, paclitaxel
• the combination of ramucirumab and paclitaxel is a cancer treatment method that is mainly treated as a second line for patients with gastric cancer who cannot be resected. It has also been reported to be useful for patients with metastatic gastric cancer.
• a combination study of ramucirumab and paclitaxel (RAINBOW study) in patients with gastroesophageal junction or gastric adenocarcinoma, the median overall survival of patients in the paclitaxel monotherapy group was 7.4 months (median progression-free survival was 7.4 months).
• the median overall survival for patients in the ramucirumab and paclitaxel group was 9.6 months (median progression-free survival was 4.4 months), compared to 2.9 months (non-patentable).
• Document 1 As shown in this result, the combination of ramucirumab and paclitaxel is recommended for the treatment of gastric cancer because it slightly improves survival time.
• Non-Patent Document 2 In recent years, for patients with unresectable or recurrent gastric / esophageal junction cancer, treatment with nivolumab, which is one of the immune checkpoint inhibitors, in addition to ramucirumab and paclitaxel has been treated with fluoropyridines and platinum. It has been reported that patients who became resistant after the first-line treatment of the drug were treated as the second-line treatment, but the 6-month progression-free survival rate was only about 46%. (Non-Patent Document 2).
• An object of the present invention is to provide a drug for treating and / or preventing a cancer that specifically expresses the CAPRIN-1 protein on the cell surface.
• the present inventor has combined an antibody against the CAPRIN-1 protein, which has immunological reactivity with cancer cells, or a fragment thereof, with an angiogenesis inhibitor, or an angiogenesis inhibitor and a taxan-based drug.
• angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug.
• they have found that they exert an extremely strong antitumor effect in cancer patients, particularly cancer patients who have a history of cancer treatment with drugs other than the combination therapy, and have completed the present invention.
• the present invention relates to the following embodiments (1) to (18).
• Treatment of cancer which comprises an antibody or fragment thereof having immunological reactivity with the CAPRIN-1 protein and an angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug in combination or separately. And / or a drug for prevention.
• angiogenesis inhibitor is an anti-VEGF inhibitor and / or an anti-vascular endothelial growth factor receptor (VEGFR) inhibitor.
• VAGFR anti-vascular endothelial growth factor receptor
• angiogenesis inhibitor is bevacizumab, ramucirumab and / or axitinib.
• a drug in which the cancer contains an antibody or fragment thereof immunoreactive with the CAPRIN-1 protein and an angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug together or separately.
• CAPRIN in which the antibody or fragment thereof has an amino acid sequence represented by any of SEQ ID NOs: 2 to 30, which is an even number, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence.
• the antibody or fragment thereof has an amino acid sequence represented by any one of SEQ ID NOs: 31 to 35, 296 to 299, 308, and 309, or an amino acid sequence having 80% or more sequence identity with the amino acid sequence.
• An antibody or fragment (F) SEQ ID NOs 176, 177 and 178 that comprises a light chain variable region containing complementarity determining regions (CDR1, CDR2 and CDR3, respectively) and is immunologically reactive with the CAPRIIN-1 protein.
• Complementarity determining regions of 185, 186 and 187 Complementarity determining of the antibodies or fragments (H) SEQ ID NOs 188, 189 and 190 which contain the light chain variable regions containing (CDR1, CDR2 and CDR3 respectively) and have immunological reactivity with the CAPRIIN-1 proteins.
• the heavy chain variable regions containing the regions (CDR1, CDR2 and CDR3, respectively) and the light chain variable regions containing the complementarity determining regions of SEQ ID NOs 191, 192 and 193 (CDR1, CDR2 and CDR3, respectively), and CAPRIIN-.
• An antibody or fragment thereof immunoreactive with a protein (I) Heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs 146, 147 and 148 (CDR1, CDR2 and CDR3, respectively) and SEQ ID NOs 149, 150 and An antibody or fragment (J) SEQ ID NOs 272, 273 that comprises a light chain variable region containing 151 complementarity determining regions (CDR1, CDR2 and CDR3, respectively) and is immunologically reactive with the CAPRIIN-1 protein. And 274 complementarity determining regions (CDR1, CDR2 and CDR3 respectively) containing heavy chain variable regions and SEQ ID NOs 275, 276 and 277 complementarity determining regions (CDR1, CDR2 and CDR3 respectively).
• A An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 39 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 43.
• B The heavy chain variable region of SEQ ID NO: 47. An antibody or fragment thereof comprising an amino acid sequence and having a light chain variable region containing the amino acid sequence of SEQ ID NO: 51.
• C A heavy chain variable region containing the amino acid sequence of SEQ ID NO: 55 and a light chain variable region.
• An antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 59
• An antibody in which the fragment (e) heavy chain variable region contains the amino acid sequence of SEQ ID NO: 68 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 69, or a fragment (f) heavy chain variable region thereof is SEQ ID NO:
• An antibody or fragment thereof (g) containing 70 amino acid sequences and the light chain variable region containing the amino acid sequence of SEQ ID NO: 71.
• the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 72 and the light chain variable.
• An antibody or fragment thereof (i) A antibody or fragment thereof (j) in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 77 is a fragment (j) heavy chain variable region thereof.
• the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 80 and is light.
• An antibody or fragment thereof in which the chain variable region comprises the amino acid sequence of SEQ ID NO: 81 (l)
• the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 82, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 83.
• Antibody or fragment thereof (m) Heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 84, and light chain variable region containing the amino acid sequence of SEQ ID NO: 85.
• An antibody or fragment thereof in which the 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, and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 88.
• Light chain variable region is the amino acid of SEQ ID NO: 89
• An antibody or fragment thereof (q) in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 90 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 91.
• the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 94.
• the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 96, and the light chain variable region is SEQ ID NO: 97.
• Antibody or fragment thereof An antibody or fragment thereof (t) in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 98 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 99 ( u) An antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 100 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 101 (v) The heavy chain variable region is the amino acid of SEQ ID NO: 102. An antibody or fragment thereof comprising a sequence and the light chain variable region containing the amino acid sequence of SEQ ID NO: 103 (w) The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 104, and the light chain variable region is a sequence.
• Antibody or fragment thereof comprising the amino acid sequence of No. 105
• An antibody in which the fragment (y) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 108 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 109 or a fragment (z) heavy chain variable region thereof is SEQ ID NO: 110.
• An antibody comprising the amino acid sequence of SEQ ID NO: 113 or a fragment thereof (ab) An antibody in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 114 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 115.
• an antibody or a fragment (ad) heavy chain thereof in which the fragment (ac) heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 116 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 117 is possible.
• An antibody or fragment (ae) of which the variant 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 contains the amino acid sequence of SEQ ID NO: 120.
• An antibody or fragment thereof (af) in which the light chain variable region contains the amino acid sequence of SEQ ID NO: 121 The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 122, and the light chain variable region is the amino acid of SEQ ID NO: 123.
• an antibody or fragment thereof in which the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 126 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 127.
• the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 128.
• Antibody or fragment thereof (ak) containing the amino acid sequence of SEQ ID NO: 132
• the antibody or fragment thereof comprising the amino acid sequence of SEQ ID NO: 132 and the light chain variable region containing the amino acid sequence of SEQ ID NO: 133 ( al)
• the cancers are gastric cancer, breast cancer, renal cancer, pancreatic cancer, colon cancer, bile duct cancer, melanoma, lung cancer, renal cell cancer, hodgkin lymphoma, head and neck cancer, mesenteric cancer, colon / rectal cancer, esophageal cancer, Gastroesophageal junction cancer, hepatocellular carcinoma, glioblastoma, urinary epithelial cancer, ovarian cancer, bladder cancer, uterine cancer, central nervous system primary lymphoma, testicular primary lymphoma, biliary tract cancer, brain tumor, prostate cancer, leukemia, lymphoma, Liver cancer, sarcoma, fibrosarcoma, obesity cell tumor, adrenal cortex cancer, Ewing tumor, multiple myeloma, testicle cancer, thyroid cancer, basal cell cancer, Paget's disease or skin cancer, (1)-(14) The drug listed in either.
• the combined use of the antibody against the CAPRIN-1 protein or a fragment thereof according to the present invention with an angiogenesis inhibitor or an angiogenesis inhibitor and a taxane-based agent is an antibody against the CAPRIIN-1 protein alone or an existing chemotherapeutic agent (angiogenesis inhibitor). It exerts a stronger antitumor effect than the combined use of a drug and a taxane drug). Furthermore, the combined use of the antibody against the CAPRIN-1 protein or a fragment thereof according to the present invention and an angiogenesis inhibitor or a drug containing an angiogenesis inhibitor and a taxan-based drug is a single combination of the existing anticancer drug therapy and the antibody against the CAPRIN-1 protein. Shows a stronger antitumor effect compared to treatment. Therefore, the combined use of an antibody against the CAPRIN-1 protein or a fragment thereof and an angiogenesis inhibitor, or an angiogenesis inhibitor and a taxane drug is effective for the treatment or prevention of cancer.
• anti-CAPRIN-1 antibody An antibody against the CAPRIN-1 protein used in the present invention or a fragment thereof (hereinafter, referred to as "anti-CAPRIN-1 antibody"), an angiogenesis inhibitor or an angiogenesis inhibitor, and a taxan-based drug (hereinafter, these are collectively referred to).
• anti-CAPRIN-1 antibody an antibody against the CAPRIN-1 protein used in the present invention or a fragment thereof
• an angiogenesis inhibitor or an angiogenesis inhibitor an angiogenesis inhibitor
• taxan-based drug hereinafter, these are collectively referred to.
• the anti-tumor activity when used in combination with an angiogenesis inhibitor or the like) can be evaluated by examining the suppression of tumor growth in a cancer-bearing animal in vivo as described later.
• “combination” or “combination” means that an anti-CAPRIN-1 antibody and an angiogenesis inhibitor or the like are administered to the same living body simultaneously or at predetermined intervals as independent active ingredients. Point to. The interval may be simultaneous administration, 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, 2 weeks, 3 It may be after a week or four weeks.
• an anti-CAPRIN-1 antibody, an angiogenesis inhibitor, or the like exhibits its activity in vivo, either of them may be administered. Further, the anti-CAPRIN-1 antibody may be administered first, or an angiogenesis inhibitor or the like may be administered first.
• the term "comprising together or separately” has the meaning that multiple agents are included in a form that can be administered to a patient simultaneously or separately.
• it may be in the form of a so-called mixed preparation in which a plurality of drugs are mixed, or in the form of a so-called kit preparation (pharmaceutical kit) in which a plurality of drugs are contained as individual preparations.
• kit preparation pharmaceutical kit
• the form also includes the form of a kit formulation, which comprises the plurality of agents in any combination of two or more formulations.
• kit preparation is, for example, a kit preparation containing a preparation (or pharmaceutical composition) containing an anti-CAPRIN-1 antibody and a preparation (or pharmaceutical composition) containing an angiogenesis inhibitor. You may.
• a preparation containing an anti-CAPRIN-1 antibody (or a pharmaceutical composition) a preparation containing an angiogenesis inhibitor (or a pharmaceutical composition), and a preparation containing a taxane-based drug. It may be a kit preparation containing (or a pharmaceutical composition).
• kit preparation As another example of the kit preparation according to the present invention, (i) a preparation (or pharmaceutical composition) containing an anti-CAPLIN-1 antibody and an angiogenesis inhibitor, and / or (ii) an anti-CAPLIN-1 antibody and a taxane system. It may be a kit preparation containing a preparation (or pharmaceutical composition) containing a drug, and (iii) a preparation (or pharmaceutical composition) containing other active ingredients.
• the anti-CAPRIN-1 antibody according to the present invention may be a monoclonal antibody or a polyclonal antibody, preferably a monoclonal antibody, and is any kind of antibody as long as the antibody of the present invention can exhibit antitumor activity.
• the antibody may be a recombinant antibody, a human antibody, a humanized antibody, a chimeric antibody, or a non-human animal antibody.
• the subject to be treated and / or prevented from cancer in the present invention is a mammal such as a human, a pet animal, a domestic animal, or a competition animal, and a preferable subject is a human.
• CAPRIN-1 having an amino acid sequence represented by any of even SEQ ID NOs: SEQ ID NOs: 2 to 30, which is a specific example of an antigen having immunological reactivity with the anti-CAPRIN-1 antibody used in the present invention.
• the amino acid sequences shown by SEQ ID NOs: 6, 8, 10, 12 and 14 are the amino acid sequences of the canine CAPRIN-1 protein
• the amino acid sequences shown by SEQ ID NOs: 2 and 4 are the human CAPRIN-1 proteins.
• the amino acid sequence shown in SEQ ID NO: 16 is the amino acid sequence of bovine CAPRIN-1 protein
• the amino acid sequence shown in SEQ ID NO: 18 is the amino acid sequence of horse CAPRIN-1 protein.
• the amino acid sequences shown by 20, 22, 24, 26 and 28 are the amino acid sequences of the mouse CAPRIN-1 protein
• the amino acid sequences shown by SEQ ID NO: 30 are the amino acid sequences of the chicken CAPRIN-1 protein.
• the anti-CAPRIN-1 antibody used in the present invention is 80% or more, preferably 90% or more, more preferably 95% or more, preferably 90% or more, more preferably 95% or more, with respect to the amino acid sequence represented by any of the even SEQ ID NOs of SEQ ID NOs: 2 to 30. It may be immunologically reactive with a variant of the CAPRIN-1 protein having a sequence identity of% or more, more preferably 99% or more.
• % sequence identity refers to the amino acid (or base) when two sequences are aligned so as to have the maximum similarity with or without a gap. It means the percentage (%) of the same amino acid (or base) to the total number.
• the anti-CAPRIN-1 antibody means an antibody having immunological reactivity with the full length of the CAPRIN-1 protein or a fragment thereof or a fragment thereof (antigen binding fragment).
• immunological reactivity means a property in which an antibody and a CAPRIN-1 protein or a partial polypeptide thereof specifically bind to each other in vivo.
• the anti-CAPRIN-1 antibody used in the present invention may be a monoclonal antibody or a polyclonal antibody.
• a polyclonal antibody (anti-CAPRIN-1 polyclonal antibody) having immunological reactivity with the full length of the CAPRIN-1 protein or a fragment thereof is, for example, a natural CAPRIN-1 protein, a fusion protein with GST, or a partial peptide thereof.
• serum After immunizing mice, human antibody-producing mice, rats, rabbits, chickens, etc., serum is obtained, and the obtained serum is subjected to ammonium sulfate precipitation, protein A, protein G, DEAE ion exchange column, CAPRIN-1 protein and a portion. It can be obtained by purifying with an affinity column or the like to which a peptide is bound.
• the nucleotide sequence and amino acid sequence of CAPRIN-1 and its homologue used for the above immunization can be obtained by accessing, for example, GenBank (NCBI, USA) and using algorithms such as BLAST and FASTA (Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90: 5873-5877, 1993; Altschul et al., Nuclear Acids Res. 25: 3389-3402, 1997). Further, the method for producing the CAPRIN-1 protein can be obtained by referring to WO2014 / 012479, and cells expressing the CAPRIIN-1 protein or the like can also be used.
• Monoclonal antibodies that have immunological reactivity with the full length of the CAPRIN-1 protein or fragments thereof are, for example, those of the breast cancer cells SK-BR-3 and CAPRIN-1 proteins that express CAPRIN-1.
• a mouse is immunized by administering the full length or a fragment thereof to the mouse, the spleen cells and myeloma cells isolated from the mouse are fused, and a clone that produces an anti-CAPRIN-1 monoclonal antibody is selected from the obtained fused cells (hybridoma).
• the antibody produced from the selected hybridoma can be obtained by the same method as the above-mentioned method for purifying the polyclonal antibody.
• Antibodies used in the present invention include human antibodies, humanized antibodies, chimeric antibodies, and non-human animal antibodies.
• the human antibody sensitizes human lymphocytes infected with EB virus with proteins, protein-expressing cells or lysates thereof, and fuses the sensitized lymphocytes with myeloma cells such as U266 cells derived from humans to obtain the fusion.
• Antibodies that are immunologically reactive with the full length of the CAPRIN-1 protein or fragments thereof can be obtained from cells.
• a humanized antibody is a modified antibody that is also called a restored human antibody.
• Humanized antibodies are constructed by transplanting the complementarity determining regions of antibodies derived from immune animals into the complementarity determining regions of human antibodies.
• the gene recombination method as the general method is also a well-known technique. Specifically, for example, several DNA sequences designed to link the complementarity determining regions of mouse antibody and rabbit antibody and the framework regions of human antibody are prepared so as to have an overlapping portion at the terminal portion. Synthesize from the oligonucleotides of the above by the PCR method.
• Antibodies are heteropolyglycoproteins that usually contain at least two heavy chains and two light chains.
• the antibody is composed of two identical light chains and two identical heavy chains.
• the heavy chain has a heavy chain variable region at one end, followed by several stationary regions.
• the light chain has a light chain variable region at one end, followed by several constant regions.
• the variable regions have specific variable regions called complementarity determining regions (CDRs) that impart binding specificity to the antibody.
• CDRs complementarity determining regions
• the relatively conserved portion of the variable region is called the framework region (FR).
• the variable regions of the complete heavy and light chains each contain four FRs linked by three CDRs (CDR1 to CDR3).
• the sequences of the constant region and variable region of human-derived heavy and light chains can be obtained from, for example, NCBI (USA: GenBank, UniGene, etc.).
• the heavy chain constant region of human IgG1 is registered in Registration No. J00228.
• the heavy chain constant region of human IgG2 refer to the sequence of registration number J00230, for the human light chain ⁇ constant region, reference to the sequences of registration numbers V00557, X64135, X64133, etc., and for the human light chain ⁇ constant region, refer to the sequences of registration numbers X64132, X64134, etc. be able to.
• a chimeric antibody is an antibody produced by combining sequences derived from different animals. For example, from the heavy chain variable region and light chain variable region of a mouse antibody and the constant region of a heavy chain variable region and a light chain variable region of a human antibody. Antibodies and the like.
• the chimeric antibody can be prepared by using a known method. For example, a DNA encoding the antibody V region and a DNA encoding the C region of a human antibody are ligated, and this is incorporated into an expression vector and introduced into a host. Obtained by producing.
• Non-human animal antibodies are obtained by injecting the sensitizing antigen intraperitoneally, intradermally or subcutaneously in an animal such as a mouse as a general method of immunizing an animal with a sensitizing antigen according to a known method. ..
• an appropriate amount is mixed with various adjuvants such as CFA (Freund Complete adjuvant) and administered to animals multiple times.
• CFA Cosmetic Acids Factor
• the serum was obtained, and as described above, sulfate precipitation, protein A, protein G, DEAE ion exchange column, CAPRIN-1 protein.
• Non-human animal antibodies can be obtained by purifying with an affinity column or the like to which a partial peptide is bound.
• a monoclonal antibody when obtained from a non-human animal, it can be obtained by collecting immune cells from an immunized animal and subjecting them to cell fusion with myeloma cells. Cell fusion between the immune cells and myeloma cells can be performed according to a known method (see Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46).
• the antibody used in the present invention can also be obtained as a recombinant antibody produced by cloning an antibody gene from a hybridoma, incorporating it into an appropriate vector, introducing it into a host, and producing it using a genetic recombination technique.
• Can Carl, AK Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Public in the United Kingdom by MACMILLAN PUBLISH).
• the anti-CAPRIN-1 antibody used in the present invention may be one in which an amino acid in a variable region (for example, FR) or a constant region is replaced with another amino acid.
• the amino acid substitution is one or more, for example, less than 15, less than 10, 8 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less amino acids, preferably 1 to 9 amino acids.
• the antibody should be an antibody that has the same or better binding property to the antigen and the binding affinity to the antigen than the unsubstituted antibody, and does not cause rejection when applied to humans. ..
• Amino acid substitutions are preferably conservative amino acid substitutions, which are substitutions between amino acids with similar properties such as charge, side chain, polarity and aromaticity.
• Amino acids with similar properties include, for example, basic amino acids (arginine, lysine, histidine), acidic amino acids (aspartic acid, glutamic acid), uncharged polar amino acids (glycine, asparagine, glutamine, serine, threonine, cysteine, tyrosine), non-polar amino acids. It can be classified into sex amino acids (leucine, isoleucine, alanine, valine, proline, phenylalanine, tryptophan, methionine), branched amino acids (threonine, valine, isoleucine), aromatic amino acids (phenylalanine, tyrosine, tryptophan, histidine) and the like.
• basic amino acids arginine, lysine, histidine
• acidic amino acids aspartic acid, glutamic acid
• uncharged polar amino acids glycine, asparagine, glutamine, serine, threonine, cyste
• the anti-CAPRIN-1 antibody used in the present invention can be expected to have a stronger antitumor effect when it has a high binding affinity with the CAPRIN-1 protein on the surface of cancer cells.
• the binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5 ⁇ 10 8 M -1 , at least 10 9 M -1 , and at least 5 ⁇ . 10 9 M -1 , at least 10 10 M -1 , at least 5 x 10 10 M -1 , at least 10 11 M -1 , at least 5 x 10 11 M -1 , at least 10 12 M -1 , or at least 10 13 It is desirable that it is M -1.
• the anti-CAPRIN-1 antibody used in the present invention may be chemically modified, and examples of such antibody modifications include polyethylene glycol (PEG) and anti-neoplastic compounds (eg, anti-neoplastic compounds exemplified below).
• Antibodies bound to various molecules such as tumor agents can be mentioned.
• the substance to be bound is not limited.
• Such antibody modifications can be obtained by chemically modifying the obtained antibody. These methods have already been established in this field.
• the anti-CAPRIN-1 antibody used in the present invention replaces one, two or several amino acids in the heavy chain constant region of the antibody, or N- in an N-glycosidic bond sugar chain that binds to the heavy chain constant region.
• the above may be an amino acid substitution alone, or may be a composition with an antibody to which fucose is bound.
• Antibodies in which one, two, or several amino acids in the heavy chain constant region are substituted are prepared by referring to, for example, WO2004 / 063351, WO2011 / 120135, US Pat. No. 8,388,955, WO2011 / 005481, US Pat. No. 6,737,056, WO2005 / 063351. can do.
• anti-CAPRIN-1 polyclonal antibody anti-CAPRIN-1 monoclonal antibody used in the present invention
• method for producing the antibody the purification method, and the method for producing the CAPRIN-1 protein or a partial polypeptide thereof used for immunization are described in WO2010 / 016526 and WO2011 /.
• anti-CAPRIN-1 antibody in the present invention examples include the aforementioned WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 /.
• amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4 or the amino acid sequence preferably 85% or more, more preferably 90% or more, still more preferably 95% or more, still more preferably 99% or more.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 31 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CDR2 and CDR3 an antibody or fragment thereof that comprises the light chain variable regions and is immunologically reactive with the CAPRIN-1 proteins.
• an antibody or fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 39 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 43, or the heavy chain variable region is SEQ ID NO: 70.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 33 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 32 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 34 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• An antibody or fragment thereof that contains a light chain variable region containing, and is immunologically reactive with the CAPRIN-1 protein, or complementarity determining regions of SEQ ID NOs: 176, 177, and 178 (CDR1, CDR2, and CDR3, respectively).
• CDR1, CDR2, and CDR3, respectively complementarity determining regions of SEQ ID NOs: 176, 177, and 178
• light chain variable regions containing the complementarity determining regions of SEQ ID NOs: 179, 180 and 181 CDR1, CDR2 and CDR3, respectively
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 35 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 296 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 297 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 298 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 299 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• the heavy chain variable regions containing the complementarity determining regions of SEQ ID NOs: 301, 302 and 303 (CDR1, CDR2 and CDR3, respectively) and the complementarity determining regions of SEQ ID NOs: 305, 306 and 307 (CDR1, CDR2 and CDR3, respectively) An antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or 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.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 308 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• an antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69.
• CAPRIN- having an amino acid sequence represented by SEQ ID NO: 309 or an amino acid sequence having 80% or more (preferably 85% or more, more preferably 90% or more, still more preferably 95% or more) sequence identity with the amino acid sequence.
• an antibody or fragment thereof that comprises a light chain variable region and has immunological reactivity with the CAPRIN-1 protein. More preferably, an antibody or a fragment thereof, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 68 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 69.
• the following anti-CAPRIN-1 antibody is also preferably used.
• Angiogenesis inhibitors are agents that suppress the migration and proliferation of vascular endothelial cells and inhibit angiogenesis, preferably anti-vascular endothelial growth factor (VEGF) inhibitors and / or anti-vascular endothelial growth factor receptors (VEGFR). ) It is an inhibitor.
• anti-VEGF inhibitors include bevacizumab, Ranibizumab, Lenvatinib meslylate, dibu-aflubercept (Avivercept), and anti-vascular factor.
• VEFGR Inhibitors include Ramushirumab, Axitinib, Cabozantinib, Regorafenib, Sorafenib, Sorafenib, Sorafenib Cabozantinib can be mentioned.
• the angiogenesis inhibitor may preferably be bevacizumab, ramucirumab, and / or axitinib.
• Taxanes are any compound that inhibits cell division microtubule depolymerization and causes microtubule hyperformation to inhibit cell division and exert an antitumor effect. May be good.
• Taxole one of the taxanes, is a tetracyclic diterpene compound in which the oxetane ring is bound to the C ring of the tricyclic taxane skeleton, and the A ring and C ring are bent so as to face each other with the B ring in between.
• Taxane-based agents include paclitaxel, docetaxel, larotaxel, nab-paclitaxel, and derivatives thereof, which are preferably used in the present invention.
• a drug containing the anti-CAPRIN-1 antibody, an angiogenesis inhibitor and a taxane drug as an active ingredient of the drug of the present invention may be used in combination with an antitumor agent known in the literature and the like.
• the known antitumor agent is not particularly limited, and specific examples thereof include padoxorubicin, daunorubicin, cyclophosphamide, methotrexate, thiotepa, busulfan, improsulfan, piposulfan, benzodopa, carbocon, and meturedopa.
• the combination of the anti-CAPRIN-1 antibody of the present invention and an angiogenesis inhibitor or the like has cytotoxic activity in vivo. Therefore, the antitumor effect of the present invention can be known by examining the cytotoxic activity against cancer. For cytotoxic activity, administer a drug containing an anti-CAPRIN-1 antibody and an angiogenesis inhibitor to a living body having cancer, measure the size of the tumor after administration, and examine the size of the cancer over time. Can be evaluated by.
• the antitumor effect of the present invention can also be evaluated by examining the survival rate. It can also be evaluated by examining the ability to produce cytokines or chemokines.
• the antitumor effect of the combination can be further investigated by examining the prevention of cancer, the prevention of metastasis, or the prevention of recurrence.
• the anti-CAPRIN-1 antibody used in the present invention can be expected to have a stronger antitumor effect when the binding affinity with the CAPRIN-1 protein on the surface of cancer cells is high.
• the binding constant (affinity constant) Ka (kon / koff) is preferably at least 10 7 M -1 , at least 10 8 M -1 , at least 5 ⁇ 10 8 M -1 , at least 10 9 M -1 , and at least 5 ⁇ . 10 9 M -1 , at least 10 10 M -1 , at least 5 x 10 10 M -1 , at least 10 11 M -1 , at least 5 x 10 11 M -1 , at least 10 12 M -1 , or at least 10 13 It is desirable that it is M -1.
• the ability of the anti-CAPRIN-1 antibody used in the present invention to bind to CAPRIN-1 can be identified using, for example, a binding assay using ELISA, Western blotting, immunofluorescence and flow cytometry. can.
• the combination of the anti-CAPRIN-1 antibody of the present invention to an angiogenesis inhibitor or the like in vivo of cancer enhances the antitumor effect as described above, but the enhancement rate is higher than that of the anti-CAPRIN-1 antibody alone.
• the enhancement rate of the antitumor effect of the combination administration of the anti-CAPRIN-1 antibody and the angiogenesis inhibitor according to the present invention with respect to the administration of the anti-CAPRIN-1 antibody alone is such that an effective amount of each is administered to the cancer-bearing mouse under the same conditions.
• the pharmaceutical product of the present invention is intended for the treatment and / or prevention of cancer.
• the cancer targeted by the drug of the present invention is not particularly limited as long as it is a cancer (cell) expressing the CAPRIN-1 protein.
• treatment means the treatment of cancer based on the aforementioned antitumor effect. Further, as used herein, prevention means not only prevention of cancer development but also prevention of cancer metastasis or recurrence.
• tumor and cancer used herein mean malignant neoplasms and are used interchangeably.
• the target cancer patients in the present invention are not particularly limited, but preferably, an antibody having immunoreactivity with the CAPRIN-1 protein or a fragment thereof, an angiogenesis inhibitor, or an angiogenesis inhibitor and a taxan-based drug.
• a cancer patient who has been treated for cancer treatment in accordance with "NCCN Clinical Practice Guidelines in Oncology", "ESMO Clinical Practice Guidelines” or “Cancer Practice Guidelines” can be mentioned. It is preferably a cancer patient having a history of cancer treatment with an angiogenesis inhibitor and / or a taxane.
• cancer by a drug other than cancer treatment by a drug containing an anti-CAPRIN-1 antibody and an angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug together or separately. It is a cancer patient who has not responded to the treatment, and more preferably a cancer patient who has not responded to the cancer treatment with an angiogenesis inhibitor and / or a taxan drug.
• cancer by a drug other than cancer treatment by a drug containing an anti-CAPRIN-1 antibody and an angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug together or separately.
• a drug containing an anti-CAPRIN-1 antibody and an angiogenesis inhibitor or an angiogenesis inhibitor and a taxan-based drug together or separately.
• a cancer patient having a cancer resistant to treatment and more preferably a cancer patient having a cancer resistant to cancer treatment with an angiogenesis inhibitor and / or a taxan drug.
• cancer treatment did not respond and "resisting to cancer treatment” are used synonymously.
• the target cancer in the present invention may be any cancer as long as it expresses the CAPRIN-1 protein on the cell membrane surface.
• these cancers may be primary cancers, metastatic cancers, metastatic or recurrent cancers, postoperative cancers, or unresectable cancers.
• melanoma is often used synonymously with malignant melanoma or malignant melanoma.
• a cancer having resistance to a known treatment method can be mentioned.
• the resistant cancer is not particularly limited as long as it is a cancer derived from a patient with any treatment history, but for example, a cancer derived from a patient who has been treated with 5-FU and becomes resistant after administration. Cancer that has, has metastasized, or has recurred.
• the cancers include, for example, Bowen's disease, melanoma, spinous cell cancer, extramammary Paget's disease, bacteriolytic sarcoma, Sezary syndrome, skin T / NK cell lymphoma, and T cells having lesions only in the skin.
• Leukemia / lymphoma cutaneous B-cell lymphoma (independent group), cutaneous T-cell lympholine cancer, complex mammary adenocarcinoma, malignant mixed mammary gland tumor, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, Large cell cancer, neuroepithelial tissue tumors such as glioma, glioblastoma, neuroblastoma, ventricular lining tumor, neurocellular tumor, fetal neuroemphatic tumor, nerve sheath tumor, neurofibroma, Meningitis, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, digestive lymphoma, small to medium cell lymphoma, cecum cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, sigmoid colon cancer, Rectal cancer, ovarian epithelial cancer, embryonic cell tumor, stromal cell tumor, pancreatic duct cancer
• palpable cancers originating from the above cancers cancers existing under the skin, cancers existing in the skin, superficial cancers, cancers existing in the dermis or cancers existing in non-parenchymal organs, and advanced cancers Included.
• metastasis or recurrence of the above cancer it is present in palpable cancer, subcutaneous cancer, intradermal cancer, superficial cancer, dermis cancer or non-parenchymal organ. Cancer is included.
• the preferred subject (patient) to be a target is a mammal, for example, a mammal including a primate, a pet animal, a domestic animal, a competition animal, etc., and a human, a dog, and a cat are particularly preferable.
• the pharmaceutical product of the present invention can be formulated by a method known to those skilled in the art.
• the pharmaceutical product of the present invention can be used parenterally in the form of, for example, a sterile solution in water or other pharmaceutically acceptable liquid, or an injection of a suspension.
• the active ingredient (at least one of the anti-CAPRIN-1 antibody, the angiogenesis inhibitor and the taxan-based agent of the present invention) is, for example, pharmacologically acceptable for each pharmaceutical product or pharmaceutical composition.
• Carrier, vehicle, or additive specifically, sterilized water, physiological saline, isotonic solution, buffer (buffer solution, etc.), vegetable oil, oily solution, antioxidant, solubilizer, emulsifier, suspension.
• turbinants turbinants, surfactants, stabilizers, fragrances, excipients, binders, etc.
• turbinants surfactants, stabilizers, fragrances, excipients, binders, etc.
• the amount of active ingredient in these formulations is such that an appropriate dose in the indicated range can be obtained.
• the sterile composition for injection can be formulated according to the usual formulation practice using a vehicle such as distilled water for injection.
• Aqueous solutions for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, such as D-sorbitol, D-mannose, D-mannitol, sodium chloride, and suitable solubilizers such as.
• Alcohols, specifically ethanol, polyalcohols such as propylene glycol, polyethylene glycol and nonionic surfactants such as polysorbate 80 (TM), HCO-60 may be used in combination.
• examples of the oily liquid include sesame oil and soybean oil, and benzyl benzoate and benzyl alcohol may be used in combination as solubilizing agents.
• buffers such as phosphate buffers, sodium acetate buffers, soothing agents such as prokine hydrochloride, stabilizers such as benzyl alcohol, phenols and antioxidants.
• the prepared injection solution is usually filled in a suitable ampoule.
• the administration is oral or parenteral, preferably parenteral administration, and specific examples thereof include injection type, nasal administration type, pulmonary administration type, and transdermal administration type.
• injection type for example, it can be administered systemically or locally by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, intratumoral injection and the like.
• transdermal administration type are, for example, those called coating agents or external preparations.
• external preparations include solid preparations, liquid preparations, spray preparations, ointments, cream preparations, and gel preparations.
• the administration method can be appropriately selected according to the patient's age, weight, gender, symptoms, and the like.
• the dose of the pharmaceutical composition containing at least one of the anti-CAPRIN-1 antibody, angiogenesis inhibitor and taxane drug is the amount of each active ingredient, for example, from 0.0001 mg per kg of body weight at a time. It is possible to select in the range of 1000 mg.
• the dosage of each active ingredient can be selected, for example, in the range of 0.001 to 100,000 mg / body per patient, or, for example, 0.1 mg to 300 mg, or 1 mg to 30 mg per kg of patient body weight. It is not always limited to the numerical value of.
• the dose and administration method vary depending on the weight, age, gender, symptom, etc. of the patient, but those skilled in the art can appropriately select the dose.
• the treatment and / or prevention of cancer with a drug for treating and / or preventing cancer of the present invention includes various forms in addition to the administration as the above-mentioned drug.
• each active ingredient of the pharmaceutical product of the present invention can be administered simultaneously, in parallel or individually in order.
• the second active ingredient can be administered within a time interval of up to about 3 weeks, that is, from immediately after the administration of the first active ingredient to about 3 weeks, and the second active ingredient can be administered.
• the third active ingredient can be administered from immediately after administration to about 3 weeks.
• the surgical procedure may be performed following the surgical procedure, or the surgical procedure may be performed between the administration of the first drug and the second drug, or between the administration of the second drug and the third drug.
• the drug for treating and / or preventing cancer of the present invention may be administered according to a plurality of administration cycles.
• the active ingredients of the pharmaceutical products for the treatment and / or prevention of the cancer of the present invention are co-administered
• the active ingredients of the present invention anti-CAPRIN-1 antibody and angiogenesis inhibitor, or angiogenesis inhibitor
• the pharmaceutical composition containing the agent and the taxan-based agent is administered as one cycle for about 2 days to about 3 weeks. After that, it can be repeated as needed according to the judgment of the doctor in charge of the treatment cycle.
• the duration of administration of each individual drug should be adjusted to span the same duration. The interval between cycles can be varied from 0 to 2 months.
• the dose of each active ingredient of the pharmaceutical product for treating and / or preventing the cancer of the present invention can be set in the same manner as the dose of each active ingredient in the above-mentioned pharmaceutical composition.
• the drug for treating and / or preventing cancer of the present invention may be in the form of a pharmaceutical kit.
• a pharmaceutical kit is a package for using an active ingredient in the form of a separate pharmaceutical composition (formulation) in a method for treating and / or preventing cancer, and the package is for administering each active ingredient. Instructions may be included.
• Each active ingredient of the above-mentioned pharmaceutical composition for treating and / or preventing cancer contained in a pharmaceutical kit is a pharmaceutical composition formulated as described above so that each active ingredient can be administered together or separately. It can be a form.
• the pharmaceutical kit contains a sufficient amount of the active ingredient in one or more doses so that each active ingredient can be administered according to the above-mentioned administration method.
• the present invention comprises the above-mentioned drug of the present invention or the anti-CAPRIN-1 antibody of the present invention and an angiogenesis inhibitor, or an angiogenesis inhibitor and a taxan-based drug.
• the present invention further provides a method for treating and / or preventing cancer, which comprises administering the above-mentioned pharmaceutical product or the like of the present invention to a subject (patient) who has or is suspected of having cancer. offer.
• anti-CAPRIN-1 antibody of the present invention in addition to the anti-CAPRIN-1 antibody of the present invention and an angiogenesis inhibitor, or angiogenesis inhibitor and taxane-based drug, other anti-tumor agents (known anti-tumor agents, etc.) are used as subjects (such as known anti-tumor agents). It may be administered to the patient). Further, in the embodiment thereof, for example, the anti-CAPRIN-1 antibody (antibody or fragment thereof) of the present invention, an angiogenesis inhibitor, an angiogenesis inhibitor and a taxane-based agent, and optionally an antitumor agent, which are contained in the above-mentioned pharmaceutical products. Can be administered to the subject (patient) simultaneously or separately.
• Example 1 Preparation of anti-CAPRIN-1 antibody
• the anti-CAPRIN-1 antibody having immunological reactivity with the CAPRIN-1 protein used in the present invention the one prepared as follows was used.
• Each spleen removed 3 days after the last immunization was sandwiched between two sterilized slide glasses, ground, washed with PBS (-) (manufactured by Nissui Co., Ltd.), and centrifuged at 1500 rpm for 10 minutes to prepare the supernatant. The removal operation was repeated 3 times to obtain spleen cells.
• the obtained spleen cells and mouse myeloma cells SP2 / 0 purchasedd from ATCC
• a PEG solution prepared by mixing 800 ⁇ l was added and allowed to stand for 5 minutes for cell fusion.
• the cells were suspended in 150 ml of RPMI1640 medium (HAT selective medium) containing 15% FBS containing 2% equivalent of Gibco's HAT solution, and a 96-well plate (Nunk). 100 ⁇ l per well of (manufactured by) was sown on 15 plates.
• RPMI1640 medium HAT selective medium
• FBS FBS containing 2% equivalent of Gibco's HAT solution
• Nunk 96-well plate
• hybridomas producing an antibody having a high absorbance value were selected.
• the selected hybridomas were added to the plates so as to be 0.5 per well of the 96-well plate and cultured. After 1 week, hybridomas forming a single colony were observed in the wells. The cells in these wells were further cultured, and hybridomas were selected based on the binding affinity of the antibody produced by the cloned hybridoma to the CAPRIN-1 protein. 1 ⁇ g / ml of CAPRIN-1 protein solution was added to 1 well of a 96-well plate, and the mixture was allowed to stand at 4 ° C. for 18 hours.
• the CDRs 1 to 3 of the heavy chain variable region of the selected antibody were identified, the base sequence was designed so that the framework region could express the heavy chain variable region containing the human antibody sequence, and the heavy chain determination of human IgG1 was performed. It was inserted into a mammalian expression vector into which a normal region was inserted. Similarly, CDRs 1 to 3 of the light chain variable region were identified, and the base sequence was designed so that the framework region could express the light chain variable region containing the sequence of the human antibody, and this was used as the light chain constant of human IgG1. It was inserted into a mammalian expression vector into which the region was inserted. The above two recombinant expression vectors were introduced into mammalian cells according to a conventional method to obtain a culture supernatant containing a humanized monoclonal antibody # 1 (humanized antibody # 1) against CAPRIN-1.
• the obtained culture supernatant containing the humanized anti-CAPRIN-1 monoclonal antibody # 1 was purified by a conventional method using Hitrap Protein A Sepharose FF (manufactured by GE Healthcare), replaced with PBS (-), and 0.22 ⁇ m. A filter (manufactured by Millipore) was prepared.
• the specific reactivity of the above-mentioned anti-CAPRIN-1 antibody with the CAPRIN-1 protein was confirmed by detecting the CAPRIN-1 protein on a plate in a solid phase using the ELISA method.
• Human cancer cells whose expression of the CAPRIN-1 gene has been confirmed by the flow cytometry method breast cancer cells (BT-474), colon cancer cells (HT-29), lung cancer cells (QG56, H1650), gastric cancer Cells (NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal cancer Cells (Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), bile sac cancer cells (TGBC14TKB), For any of fibrosarcoma cells (HT-1080), melanoma cells (G-361), mouse renal cancer cells (Renca) and mouse breast cancer cells (4T1) whose expression of the CAPRIN-1 gene has been confirmed.
• fibrosarcoma cells
• humanized antibody # 1 has stronger fluorescence intensity than human IgG control antibody and rabbit IgG antibody, which are not reactive with cancer cells, which are negative controls, and the above-mentioned cancer cells expressing CAPRIN-1. It was confirmed that it reacts strongly with the cell membrane surface of.
• Example 2 Anti-tumor effect by combined use of anti-CAPRIN-1 antibody and angiogenesis inhibitor in a human cancer cell-bearing mouse model Anti-CAPRIN-1 antibody (anti-CAPRIN-1 humanized antibody) prepared in Example 1. The antitumor effect of the combination of # 1) and the angiogenesis inhibitor bevasizumab (anti-VEGF antibody) was evaluated in vivo in cancer-bearing mice.
• BT474 is a cancer cell in which the CAPRIN-1 protein is expressed on the cell membrane surface, and it has been confirmed that the anti-CAPRIN-1 antibody prepared in Example 1 reacts with a part of CAPRIN-1 on the cell membrane surface. ..
• the anti-CAPRIN-1 antibody prepared in Example 1 was administered to the tail veins of the above-mentioned cancer-bearing mice at 10 mg / kg once a week. To the same mice, bevacizumab was started once a week at 1.25 mg / kg at the same time as the first administration of the anti-CAPRIN-1 antibody.
• the same anti-CAPRIN-1 antibody as above was administered to cancer-bearing mice in the same amount once a week.
• bevacizumab was administered to another cancer-bearing mouse individual at the same administration interval and dose.
• cancer-bearing mice in the untreated group were used as negative controls.
• the size of the cancer in the cancer-bearing mouse was measured over time using a caliper, and the tumor volume was calculated according to the formula: (length of the major axis of the cancer) ⁇ (length of the minor axis of the cancer). ) Calculated as 2 ⁇ 0.5.
• the tumor volume of the anti-CAPRIN-1 humanized antibody # 1 administration group prepared in Example 1 which was the comparison group was It was 52% and about 91% in the bevasizumab-treated group.
• the tumor volume was about 14%, and the tumor volume was smaller than that at the start of administration.
• anti-CAPRIN-1 humanized antibody # 1 prepared in Example 1 in combination with an anti-vascular endothelial growth factor receptor (VEGFR) inhibitor.
• VEGFR anti-vascular endothelial growth factor receptor
• the antitumor effect of the combined use of the anti-CAPRIN-1 antibody and axitinib in the present invention was examined using NOG mice in which human-derived cancer cells expressing the CAPRIN-1 protein were subcutaneously transplanted. bottom.
• the per mouse 2 ⁇ 10 7 cells of the human breast cancer cells BT474 were mixed with Matrigel and implanted subcutaneously were produced tumor-bearing mice were grown to tumors of about 100 mm 3.
• the anti-CAPRIN-1 antibody prepared in Example 1 was administered to the tail veins of the above-mentioned cancer-bearing mice at 10 mg / kg once a week.
• Axitinib was administered to the same mice at the same time as the first administration of the anti-CAPRIN-1 antibody, at 5 or 25 mg / kg twice daily for 9 days, and then once daily for 15 days.
• the same anti-CAPRIN-1 antibody as above was administered to cancer-bearing mice in the same amount once a week.
• axitinib was administered to another cancer-bearing mouse individual at the same administration interval and dose.
• cancer-bearing mice in the untreated group were used as negative controls.
• the size of the cancer in the cancer-bearing mouse was measured over time using a caliper, and the tumor volume was calculated according to the formula: (length of the major axis of the cancer) ⁇ (length of the minor axis of the cancer). ) Calculated as 2 ⁇ 0.5.
• the tumor volume of the anti-CAPRIN-1 humanized antibody # 1 administration group prepared in Example 1 which was the comparison group was It was 40%, 39% in the 5 mg / kg axitinib group, and 27% in the 25 mg / kg axitinib group.
• the group to which the humanized antibody # 1 prepared in Example 1 and 5 mg / kg axitinib were co-administered was 18%, and the group to which 25 mg / kg axitinib was co-administered was 8%, and the tumor volume was at the start of administration.
• the group to which the humanized antibody # 1 prepared in Example 1 and 5 mg / kg axitinib were co-administered was 18%, and the group to which 25 mg / kg axitinib was co-administered was 8%, and the tumor volume was at the start of administration.
• the anti-CAPRIN-1 antibody was significantly administered alone and the axitinib was administered alone. It has been shown to have a strong antitumor effect.
• Example 3 Antitumor effect by combined use of anti-CAPRIN-1 antibody with angiogenesis inhibitor and taxan-based drug Existing first-line treatment (Cisplatin, 5-FU (fluorouracil), Levofolinate, trastuzumab (Trastuzumab), Capecitabine, Taxotere, FOLFOX (combination therapy with fluorouracil, phoric acid (levofolinate) and oxaliplatin), FOLFIRI (combination therapy with fluorouracil, phoric acid (levoholinate) and oxaliplatin) ) Or Oxaliplatin), etc., and for 6 patients with gastric cancer who did not respond sufficiently to these treatments, paclitaxel, which is a taxan drug, and anti-VEGFR antibody (ramsylumab), which is an angiogenesis inhibitor.
• paclitaxel which is a taxan drug
• anti-VEGFR antibody ramsylumab

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• 2021
  • 2021-03-11 JP JP2021532006A patent/JP7797875B2/ja active Active
  • 2021-03-11 WO PCT/JP2021/009798 patent/WO2021182573A1/ja not_active Ceased
  • 2021-03-11 KR KR1020227035364A patent/KR20220152318A/ko active Pending
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  • 2021-03-11 US US17/910,538 patent/US20230129035A1/en active Pending
• 2025
  • 2025-03-27 JP JP2025053695A patent/JP2025092589A/ja active Pending

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