WO2012105219A1 - Antibody therapy effect-enhancing drug - Google Patents

Antibody therapy effect-enhancing drug Download PDF

Info

Publication number
WO2012105219A1
WO2012105219A1 PCT/JP2012/000579 JP2012000579W WO2012105219A1 WO 2012105219 A1 WO2012105219 A1 WO 2012105219A1 JP 2012000579 W JP2012000579 W JP 2012000579W WO 2012105219 A1 WO2012105219 A1 WO 2012105219A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
tcp
cells
tricalcium phosphate
antibody therapy
Prior art date
Application number
PCT/JP2012/000579
Other languages
French (fr)
Japanese (ja)
Inventor
丸山　宏二
満 高橋
雅裕 中川
靖人 秋山
秀衛 石井
錦雁 程
Original Assignee
オリンパス株式会社
静岡県
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by オリンパス株式会社, 静岡県 filed Critical オリンパス株式会社
Priority to JP2012555739A priority Critical patent/JP5990752B2/en
Publication of WO2012105219A1 publication Critical patent/WO2012105219A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/42—Phosphorus; Compounds thereof
    • 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
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
    • 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
    • C07K16/28—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
    • C07K16/3007—Carcino-embryonic Antigens
    • 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
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • 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

Definitions

  • the present invention relates to an antibody therapy effect enhancer (antibody therapy adjuvant) or antibody therapy kit, more specifically, ⁇ -tricalcium phosphate (TCP) used as an active ingredient after transplanted into the body of a subject.
  • an antibody therapy kit comprising an antibody pharmaceutical composition comprising a composition containing ⁇ -tricalcium phosphate and an antibody pharmaceutical composition.
  • the conventional treatment with chemotherapy has the problem that the abnormal cells affected by the disease and the normal cells are equally affected, thereby causing strong side effects. Therefore, development of molecular target drugs that exert effects only on abnormal cells has been underway.
  • antibody therapy using antibody drugs does not affect normal cells as much as possible by using an antibody against an antigen that is not expressed in normal cells or is expressed in very small amounts. A highly selective therapeutic effect is expected for abnormal cells with few side effects.
  • Cancer cells for example, promote cell proliferation, which can be said to be a biological characteristic of cancer, suppression of apoptosis, angiogenesis, metastasis, etc. due to deletion or abnormality of genes that play an important role in normal cell growth, differentiation, and cell death Indicates.
  • Genes related to these have been identified as oncogenes and tumor suppressor genes.
  • cancer can be treated by suppressing the functions of oncogenes.
  • administration of antibodies targeting growth factors and growth factor receptors that cause cancer has little effect on normal cells, suppresses the growth signal of cancer cells and suppresses growth or It is believed that it can be stopped.
  • a monoclonal antibody against epidermal growth factor receptor type 2 (HER2; Human Epidermal Growth Factor Receptor Type2; also known as c-erbB-2), whose overexpression is observed in various cancer cells, is an anti-neoplastic agent Herceptin (HERCEPTIN (registered trademark; manufactured by Roche), a monoclonal antibody against epidermal growth factor receptor (EGFR), which is also observed to be overexpressed in various cancer cells, is an anti-neoplastic agent ERBITUX (ERBITUX). , Registered trademark; manufactured by Merck Serono Co., Ltd.).
  • Patent Documents 1 and 2 calcium phosphate compounds containing ⁇ -tricalcium phosphate have been actively studied as biomaterials such as artificial bones and artificial roots due to their excellent biocompatibility.
  • the present inventors have coated a sheet with ⁇ -tricalcium phosphate having a porosity of 75% and a particle size of 25 to 75 ⁇ m or 75 to 105 ⁇ m, and a cancer cell inhibitor that activates macrophage activity (Patent Document 3, non-patent document) Reference 1) is being developed.
  • Patent Document 3, non-patent document) Reference 1 the effect of combining ⁇ -tricalcium phosphate and an antibody drug has not been known at all.
  • JP 2010-233914 A Japanese Patent Laid-Open No. 7-116175 JP 2010-126434 A JP 2006-129839 A Japanese Patent Laid-Open No. 2004-255110 JP 2008-163015 A
  • An object of the present invention is to provide an inexpensive and safe antibody therapy adjuvant therapeutic agent that can enhance the effect of an antibody drug.
  • the inventors have disclosed T cells, B cells, NK cells, etc. that are activated, induced, and accumulated by ⁇ -tricalcium phosphate dense bodies (here, those having a porosity of 50% or less) transplanted near the lesion site.
  • Lymphocytes and dendritic cells have a highly organized three-dimensional structure that allows lymphocytes to interact with antigens and antigen-presenting cells to initiate antigen-specific immune responses (adaptive immunity)
  • the present inventors have found that a cancer treatment effect by a medicine can be enhanced and completed the present invention.
  • the present invention relates to [1] an antibody therapy kit comprising a composition containing ⁇ -tricalcium phosphate as an active ingredient transplanted and used in a subject's body, and an antibody pharmaceutical composition; [2] The antibody therapy kit according to [1], characterized by being subcutaneous, and the antibody according to [1] or [2], wherein [3] ⁇ -tricalcium phosphate is transplanted in the vicinity of a lesion A therapeutic kit, or [4] an antibody therapy kit according to [3], wherein the vicinity of the lesion is a region at a distance of 0.1 to 15 cm from the lesion; The antibody therapy kit according to any one of [1] to [4], wherein [6] the cancer-specific antibody is an anti-CEA antibody, an anti-HER2 antibody, and an anti-EGFR antibody, It is one type or two or more types of antibodies selected [5] The antibody therapy kit according to [5].
  • [7] The antibody therapy kit according to any one of [1] to [6], wherein [7] ⁇ -tricalcium phosphate is a dense body having a porosity of 50% or less; ⁇ -tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells, and macrophages, [9] the antibody therapy kit according to [7],
  • the ⁇ -tricalcium phosphate is a tablet or a columnar body, and the antibody therapy kit according to any one of [1] to [8], or [10] ⁇ -tricalcium phosphate has a particle size
  • the present invention also provides [11] an agent for enhancing the effectiveness of antibody therapy using an antibody pharmaceutical composition comprising ⁇ -tricalcium phosphate transplanted and used in a subject's body as an active ingredient, [11] or [12], wherein the antibody therapy effect-enhancing agent described in [11] or [13] ⁇ -tricalcium phosphate is transplanted in the vicinity of a lesion Antibody therapeutic effect enhancer, [14] The antibody therapeutic effect enhancer according to [13], wherein the vicinity of the lesion is a region at a distance of 0.1 to 15 cm from the lesion, [15] The antibody therapeutic effect enhancer according to any one of [11] to [14], wherein the antibody drug is a cancer-specific antibody drug, or [16] the cancer-specific antibody is an anti-CEA antibody , One selected from anti-HER2 antibody and anti-EGFR antibody Alternatively, the antibody therapy effect-enhancing agent of [15], wherein the antibody therapy effect-enhancing agent is characterized by two or more antibodies.
  • [17] The antibody therapy effect-enhancing agent according to any one of [11] to [16], wherein [17] ⁇ -tricalcium phosphate is a dense body having a porosity of 50% or less.
  • [18] ⁇ -tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells and macrophages, and enhances the effect of antibody therapy according to [17]
  • [19] ⁇ -tricalcium phosphate is a tablet or a columnar body, or the antibody therapy effect-enhancing agent according to any one of [11] to [18], or [20] ⁇ -
  • an antibody therapeutic effect enhancer comprising ⁇ -tricalcium phosphate used as an active ingredient transplanted in the body of a subject, and to reduce the effect of an antibody drug.
  • Diseases can be treated with safe and safe antibody therapy adjuvants.
  • the results of observing the binding of immune cells and ⁇ -TCP granules (size 25-75 ⁇ m) with a scanning electron microscope are shown.
  • the results of observation of a dense ⁇ -TCP (dense tablet: ⁇ 5 ⁇ 2 mm) with a scanning electron microscope are shown. Particles with a diameter of 2 ⁇ m are used as markers.
  • the result of observing a ⁇ -TCP dense body (dense rod: ⁇ 0.9 ⁇ 5 mm) with a scanning electron microscope is shown.
  • ⁇ -TCP dense bodies (tablets) were transplanted subcutaneously into normal mice, and after 2 weeks, tissue samples were stained with HE and observed with a microscope.
  • a ⁇ -TCP dense body (granule) size of 0.05 to 105 ⁇ m was transplanted subcutaneously into normal mice, and after 2 weeks, the tissue sample was stained with HE and observed with a microscope.
  • a tissue sample of a normal mouse transplanted subcutaneously with a disk-like ⁇ -TCP compact tablet ⁇ 5 ⁇ 2 mm was stained with NK cells using an anti-CD49b antibody and observed with a microscope. It is a figure which shows that a NK cell is induced
  • a tissue sample of a normal mouse transplanted subcutaneously with a disk-like ⁇ -TCP dense tablet ⁇ 5 ⁇ 2 mm was subjected to immunohistochemical staining of dendritic cells using an anti-CD11c antibody and observed with a microscope.
  • a tissue sample of a normal mouse transplanted subcutaneously with a disk-like ⁇ -TCP dense tablet ⁇ 5 ⁇ 2 mm was immunohistologically stained for lymphatic endothelial cells using an anti-D2-40 antibody and observed with a microscope. It is a figure which shows that lymphatic vessel formation is induced
  • a disk-shaped ⁇ -TCP dense tablet or a plastic piece of the same size is transplanted subcutaneously into nude mice, and simultaneously, intraperitoneal administration of anti-CEA antibody or IgG is started, and the antibody or IgG is administered twice a week for a total of 8 times.
  • anti-CEA antibody (a-CEA) or IgG was administered twice a week for a total of 8 times peritoneal cavity.
  • the results of measuring the tumor volume twice a week thereafter are shown.
  • the tumor volume of the group of anti-CEA antibody and ⁇ -TCP is the plastic strip and IgG control group (group 1), or the group of plastic strip and anti-CEA antibody (group 3). Each group) is significantly smaller by Tukey's test.
  • CTRL indicates control IgG administration
  • a-CEA indicates anti-CEA antibody. The day when the disc-like ⁇ -TCP dense tablet or plastic piece of the same size was transplanted subcutaneously into nude mice was defined as Day 0, and anti-EGFR antibody (Erbitax) or IgG was administered intraperitoneally three times from Day 0. The result of measuring the tumor volume is shown.
  • the antibody therapy kit of the present invention comprises a composition comprising ⁇ -tricalcium phosphate (hereinafter referred to as “ ⁇ -TCP”) as an active ingredient, which is used after being implanted in the body, in other words, for implantation in the body.
  • ⁇ -TCP ⁇ -tricalcium phosphate
  • the antibody therapeutic composition is not particularly limited as long as it comprises an antibody pharmaceutical composition, and the antibody therapy effect-enhancing agent using the antibody pharmaceutical composition of the present invention is a composition containing ⁇ -TCP as an active ingredient.
  • antibody therapy in which an antibody pharmaceutical composition is administered after transplanting a composition containing ⁇ -tricalcium phosphate as an active ingredient into the body of a subject, or an antibody pharmaceutical composition.
  • ⁇ -TCP is a single crystal form of a composition represented by Ca 3 (PO 4 ) 2 and is a compound that is stable at room temperature and has biocompatibility.
  • ⁇ -TCP commercially available ⁇ -TCP powder or ⁇ -TCP block, for example, a ⁇ -TCP block used as an artificial bone, or the like may be used, or it may be produced by a known method (Japanese Patent Application Laid-Open No. 2004-26648). Alternatively, it can be produced from ⁇ -TCP powder (JP 2006-89311 A).
  • the ⁇ -TCP composition of the present invention may be substantially composed of a ⁇ -TCP composition, or may be produced by mixing a surfactant, water, or the like.
  • the ⁇ -TCP powder is densified by tableting, or defoamed by sintering to be densified, and has a high strength, high density, and low porosity.
  • a ⁇ -TCP dense body, particularly a ⁇ -TCP dense body having a porosity of 50% or less is preferable.
  • the ⁇ -TCP dense body means ⁇ -TCP having a porosity of 50% or less.
  • powder, particles, and granules all indicate granular forms, and there is no substantial distinction.
  • the particle size of the polycrystalline body constituting such a ⁇ -TCP dense body is preferably 0.001 to 50 ⁇ m, more preferably 0.01 to 20 ⁇ m, still more preferably 0.75 to 10.0 ⁇ m.
  • the grain size of the crystal is preferably 1.80 ⁇ m.
  • the gap between the polycrystalline particles is preferably 0.001 to 10 ⁇ m, more preferably 0.005 to 5 ⁇ m, and 0.01 to 1.0 ⁇ m.
  • the average polycrystalline particle gap is preferably 0. .05 to 0.2 ⁇ m, more preferably 0.1 ⁇ m.
  • the pores of the ⁇ -TCP dense body are determined by the size and number of the gaps between the polycrystalline particles.
  • the porosity of ⁇ -TCP is measured from the density by applying a pressure to inject mercury into the pores and determining the specific surface area and pore distribution from the pressure and the amount of mercury injected, and a calibration curve. You can also.
  • the porosity P can also be measured using, for example, AccuPick 1330 series (manufactured by Shimadzu Corporation).
  • the porosity of the ⁇ -TCP block measured by the mercury method is preferably 50 to 90%, more preferably 60 to 85%, still more preferably 70 to 80%.
  • ⁇ -TCP powder having a particle size of 75 to 105 ⁇ m can also be prepared by pulverizing ⁇ -TCP block, and the porosity of each powder measured by mercury method is preferably 50 to 90%, more preferably Is 55 to 85%, more preferably 57 to 80%, and a preferred average porosity is 60 to 70%.
  • ⁇ -TCP micron granules having a particle size of 25 ⁇ m or less can be prepared by pulverizing ⁇ -TCP blocks into particles, and the porosity measured by the mercury method is preferably 50% or less, more preferably It is 40% or less, more preferably 30% or less, and a preferable average porosity is 3.5 to 4.5%.
  • the ⁇ -TCP dense body include ⁇ -TCP powder, particles, granules, tablets, and columnar bodies having a porosity of 50% or less, and these may be composed only of ⁇ -TCP.
  • ⁇ -TCP may be an active ingredient and other ingredients may be included.
  • the porosity of ⁇ -TCP of 50% or less is preferably 40% or less, more preferably 30% or less, and particularly preferably 20% or less.
  • the porosity calculated from the density of the ⁇ -TCP dense body obtained by densifying ⁇ -TCP powder by tableting is preferably 50% or less, more preferably 40% or less, still more preferably 30% or less, especially 20%.
  • the ⁇ -TCP dense body defoamed by sintering has a porosity calculated from the density of preferably 50% or less, more preferably 40% or less, still more preferably 30% or less, especially 20%. The following can be illustrated suitably.
  • Densification by sintering is preferably more than 600 ° C. to 2000 ° C., more preferably 750 to 1500 ° C., further preferably 900 to 1300 ° C., and preferably 1 to 100 hours, more preferably 10 to 80 hours, A preferred example is sintering for 20 to 50 hours.
  • the sintering may be performed in a plurality of times at a plurality of temperatures.
  • a ⁇ -TCP dense body of a tablet can be produced by the following method. A slurry is prepared by mixing calcium hydrogen phosphate, calcium carbonate and water in an appropriate ratio, and the prepared slurry is reacted while being ground and then dried.
  • the solid obtained by drying is pulverized and calcined to obtain ⁇ -TCP powder.
  • the obtained ⁇ -TCP powder is compressed into a columnar body.
  • the compressed columnar body is sintered at 600 ° C. to 1300 ° C. for 20 hours.
  • particles, and granules having a porosity of 50% or less for example, calcium hydrogen phosphate and calcium carbonate powder are used in a molar ratio of 1: 1 to 3, preferably 1: Weigh and mix to 1.5 to 2.5, add pure water to the calcium hydrogen phosphate-calcium carbonate mixture to prepare a slurry, and wet-polish the prepared slurry with a ball mill for about 24 hours.
  • the reaction by the mechanochemical method is promoted by pulverization treatment, the slurry subjected to the pulverization treatment is dried at 70 to 90 ° C., preferably 75 to 85 ° C., and the solid matter obtained by drying is pulverized.
  • the obtained ⁇ -TCP powder is calcined at 700 to 800 ° C. for several hours to produce a ⁇ -TCP calcined powder, and the obtained ⁇ -TCP calcined powder is sieved to obtain each particle.
  • a fraction of diameter Can be, by measuring the porosity of each fractionation, can be obtained porosity of 50% or less of the fractions. Specifically, first, the sieve passes through a 105 ⁇ m sieve, and the sieve passes through the 75 ⁇ m sieve to leave the remaining ⁇ -TCP as a fraction (A), and then the sieve passes through a 75 ⁇ m sieve.
  • fraction (B) ⁇ -TCP remaining through a sieve having a sieve size of 25 ⁇ m
  • fraction (C) ⁇ -TCP that has passed through a 25 ⁇ m sieve
  • the ⁇ -TCP calcined powder is prepared as described above, and the obtained ⁇ -TCP calcined powder is molded into tablets or columns by compression molding (tabletting).
  • the sintered ⁇ -TCP calcined tablet or columnar body is sintered at 450 to 650 ° C., preferably 600 ° C. for 2.5 to 3.5 hours, preferably 3 hours, and then 850 to 950 ° C., preferably 900 ° C.
  • the ⁇ -TCP calcined powder is sintered to obtain a powder as a sintered body of ⁇ -TCP, which is then compression-molded (tablet). TCP tablets and columns may be produced.
  • the ⁇ -TCP columnar body may be produced by punching a ⁇ -TCP tablet into a columnar shape.
  • the ⁇ -TCP composition of the present invention includes conventional pharmaceutically acceptable carriers, binders, stabilizers, excipients, diluents, pH buffering agents, disintegrants, solubilizing agents. May contain various preparation compounding ingredients such as an agent, a solubilizing agent, and an isotonic agent, and may be in the form of a gel, paste, suspension of fine particles, or a solid preparation. Examples of the shape include granules, powders, particles, disks (short cylinders), rods (long cylinders), and the like.
  • the particle diameter of the solid ⁇ -TCP composition is, for example, in the case of ⁇ -TCP powder obtained by processing a ⁇ -TCP block into particles, the particle diameter is preferably 50 to 120 ⁇ m, more preferably 60 to It is 110 ⁇ m, more preferably 75 to 105 ⁇ m. In the case of ⁇ -TCP micron granules obtained by processing ⁇ -TCP blocks into particles, the particle size is preferably 50 ⁇ m or less, more preferably 40 ⁇ m or less, and even more preferably 30 ⁇ m or less.
  • ⁇ -TCP micron granules having a particle size of 0.05 to 5 ⁇ m and ⁇ -TCP micron granules having a particle size of 0.05 to 25 ⁇ m can be preferably exemplified.
  • particles having a particle size of 0.05 to 25 ⁇ m refer to particles that have passed through a 25 ⁇ m sieve.
  • the size of the ⁇ -TCP composition of a disk-shaped or columnar or rod-shaped tablet is preferably 0.01 to 30 mm in diameter, more preferably 0.1 to 20 mm, and still more preferably 1 to 10 mm. Is preferably 0.1 to 40 mm, more preferably 0.5 to 30 mm, and still more preferably 1 to 20 mm.
  • a tablet having a diameter of 5 mm and a length of 2 mm, or a long tablet having a diameter of 0.9 mm is preferable.
  • a columnar bar having a thickness of 10 mm can be suitably exemplified.
  • the body in the present invention is not particularly limited as long as it is a site other than bones and teeth, and can include subcutaneous, intradermal, intramuscular, intraperitoneal, intrathoracic, and intracerebral, preferably subcutaneously, more preferably
  • the vicinity of the lesion to be treated can be cited. When used by transplanting in the vicinity of a lesion, it can be used by transplanting into a region at a distance of preferably 1 ⁇ m to 30 cm, more preferably 100 ⁇ m to 20 cm, still more preferably 1 mm to 15 cm from the lesion.
  • the lesion to be treated according to the present invention is not particularly limited as long as it is a lesion other than a bone or a tooth, and examples thereof include cancer, allergic immunity, infectious diseases, etc.
  • cancer can be preferably mentioned.
  • the administration method of ⁇ -TCP of the present invention is not particularly limited as long as it is a method of transplanting directly into the body by a surgical procedure, and the ⁇ -TCP of the present invention is transplanted at the incision site at the time of surgery for other purposes. You can also Further, from the viewpoint of reducing the burden on the living body, a method of transplanting the ⁇ -TCP composition of the present invention into the body from a minimum opening by using a normal syringe or a syringe for inserting a solid material can be suitably exemplified. it can.
  • the ⁇ -TCP composition of the present invention includes a suspension containing ⁇ -TCP granules, a syringe filled with ⁇ -TCP or a solid ⁇ -TCP prepared in a gel or paste form, such as a rod-like ( Examples include a device (introduction device) filled with a ⁇ -TCP dense body having a long cylindrical shape and a set of a rod-like (long cylindrical shape) ⁇ -TCP dense body and its device (introduction device).
  • the dose can be appropriately selected according to the type of disease, the size of the lesion, the weight of the patient, etc., preferably 0.01 mg to 100 g, more preferably 0.1 mg to 10 g, and still more preferably 1 mg to 1 g. Can be illustrated.
  • the antibody pharmaceutical composition in the present invention is not particularly limited as long as it is a composition containing an antibody pharmaceutical, and preferably shows an antigen specifically expressed in cancer cells or low expression in normal cells. It is a composition containing an antibody against an antigen that exhibits high expression in cells, and it can be prepared as an antibody, or a commercially available product can be obtained as an antibody or antibody pharmaceutical composition.
  • anti-CEA antibody anti-HER2 antibody
  • anti-HER2 antibody Trastuzumab: Trastuzumab, trade name Herceptin (registered trademark) (Chugai Pharmaceutical Co., Ltd.)
  • anti-EGFR antibody Arbitux: ERBITUX
  • anti-CD20 antibody rituximab: RITUXIMAB and ibritumomab
  • Anti-CD52 antibody (aletuzumab; ALEMTUZUMAB, also known as CAMPATH)
  • anti-17-1A human tumor-associated epithelial cell adhesion factor) antibody
  • anti-VEGF vascular endothelial growth factor
  • bevacizumab BEVACIZUMAB, product)
  • Avastin registered trademark
  • Genentech anti-TNF- ⁇ antibody
  • anti-TNF- ⁇ antibody adalimumab: adalimumab, trade name Humira (registered trademark) (Eisai Co., Ltd.
  • Such antibody pharmaceutical compositions include pharmaceutically acceptable ordinary carriers, binders, stabilizers, excipients, diluents, pH buffers, disintegrants, solubilizers, solubilizers, isotonic agents, and the like. It may contain various compounding ingredients and can be administered orally or parenterally. For example, it is orally administered in dosage forms such as powders, granules, tablets, capsules, syrups and suspensions. Or can be administered parenterally by injection in the form of a solution, emulsion, suspension, etc., or can be administered intranasally in the form of a spray, A parenteral administration by drip can be preferably exemplified.
  • the antibody therapy kit of the present invention or the effect enhancer of the antibody therapy of the present invention comprises: a) transplanting ⁇ -tricalcium phosphate into the body of a subject; and b) administering an antibody pharmaceutical composition to the subject. It can also be used for antibody therapy with an antibody drug provided.
  • the antibody pharmaceutical composition is administered to the subject at the same time before or after ⁇ -TCP is transplanted, and the dosage and frequency of administration are the strength of the activity of the antibody pharmaceutical, the type of disease, the weight of the patient, the dosage form, etc. For example, preferably 0.01 to 100 mg / kg (body weight) per day, more preferably 0.05 to 70 mg / kg (body weight), still more preferably 0.1 to 50 mg / kg. (Weight) can be mentioned.
  • administration of the antibody pharmaceutical composition to the subject is preferably started at the same time or after transplantation of ⁇ -TCP of the present invention, and is administered at a fixed period, preferably every 2 weeks, for a total of 8 times.
  • the antibody drug in the present invention may be used in combination with administration of other drugs and supplements other than the antibody drug, or may be used in combination with surgical therapy such as surgery.
  • the effect enhancer of the antibody therapy using the antibody pharmaceutical of the present invention is a treatment in which the antibody therapy only by administration of the antibody pharmaceutical composition and the control or administration of the ⁇ -TCP composition and the antibody pharmaceutical composition of the present invention are used in combination.
  • the combined use of the ⁇ -TCP composition has the effect of obtaining a higher therapeutic effect.
  • the tumor marker value is decreased or when the tumor size is compared as an index
  • the tumor size is not reduced or increased when ⁇ -TCP is used in addition to the antibody therapy, compared to the control and antibody therapy alone.
  • the present invention has an effect that a mouse transplanted with ⁇ -TCP can also be used for verification of the effect of a new antibody pharmaceutical composition and for screening of an antibody pharmaceutical composition having a weak drug effect at the stage of a candidate substance.
  • nude mice are often used for verification of the antitumor effect, but in nude mice, the only effector cells that may have an antitumor effect are NK cells, whereas in the experimental system of the present invention, normal Since it is possible to examine the involvement of not only NK cells but also T cells and B cells using a simple mouse, a test closer to human clinical (cancer patients) can be performed.
  • the ⁇ -TCP block can be purchased as an artificial bone, but can also be produced by mixing and sintering raw material ⁇ -TCP and water, a surfactant and a foaming agent.
  • Such ⁇ -TCP block had a porosity of 75% and a porosity of 60%.
  • the ⁇ -TCP block having a porosity of 75% is crushed and sieved to obtain a ⁇ -TCP powder having a particle size of 75 to 105 ⁇ m (porosity: 60 to 70%). Obtained.
  • ⁇ -TCP powder is obtained, and the powder is compressed and formed into a ⁇ 5 ⁇ 2 mm tablet and sintered at 600-1500 ° C. for 30 hours, so that a disk-like ⁇ -TCP dense body is obtained. Tablets were prepared (porosity: 0.1-20%).
  • ⁇ -TCP raw material powder 6.0 ml of surfactant, and 10 ml of water are poured into a centrifuge tube, defoamed by being treated at 250 rpm for 4 minutes, poured into a mold, and naturally dried. After releasing the mold, sintering was performed at 1300 ° C. to prepare a columnar ⁇ -TCP dense rod having a diameter of 0.9 mm ⁇ 10 mm (porosity: 10 to 30%). The porosity of each ⁇ -TCP produced was measured by the mercury method as a value relative to the sample volume when mercury was pressed into pores having a diameter of about 180 ⁇ m corresponding to the initial pressure.
  • FIG. 1 shows the results of observation of the binding of the prepared ⁇ -TCP granules (size 25 to 75 ⁇ m) and immune cells with a scanning electron microscope. Further, the results of observation of the produced ⁇ -TCP dense tablet and the dense rod with a scanning electron microscope are shown in FIGS.
  • FIG. 4 shows the results of tissue samples collected from normal mice transplanted subcutaneously with ⁇ -TCP dense bodies, stained with HE, and observed with a microscope. It was found that lymphocytes accumulate and have a structure similar to that of natural lymph nodes. Further, FIG.
  • ⁇ -TCP granules (porosity 0 to 70%) of size 0.05 to 105 ⁇ m, stained with HE, and observed with a microscope.
  • tissue sample of a mouse transplanted subcutaneously with ⁇ 5 ⁇ 2 mm ⁇ -TCP dense body (porosity: 18%) was used as an anti-CD45R antibody, anti-CD3 antibody, anti-CD49b antibody, anti-CD11c antibody, anti-F4 / 80R antibody, Alternatively, using an anti-D2-40 antibody, B cells, T cells, NK cells, dendritic cells, macrophages, and lymphatic vessels were immunohistologically stained and observed under a microscope. The results are shown in FIGS.
  • ⁇ -TCP densified by sintering implanted subcutaneously can induce lymphocytes (B cells, T cells, NK cells), dendritic cells, and macrophages and can be used as an artificial lymph node. confirmed.
  • Nude mice are often used for verification of antitumor effects. However, in nude mice, the only effector cells that may have an antitumor effect are NK cells, whereas mice transplanted with ⁇ -TCP of the present invention can also investigate the involvement of T cells and B cells. It was confirmed that the test can be conducted in a state closer to human clinical (cancer patients).
  • a tablet of disk-shaped ⁇ -TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor.
  • a plastic carrier material hereinafter referred to as “plastic piece” of the same size that does not contain a heterologous protein that may have immunogenicity is implanted subcutaneously and controlled. It was. The day on which ⁇ -TCP or plastic pieces were implanted was set to Day 0, and the antibody was administered intraperitoneally a total of 8 times, twice a week from Day 0.
  • the anti-CEA antibody was administered at a dose of 50 ⁇ g per dose, and mouse IgG 50 ⁇ g was administered as a control in groups 1 and 2 where no anti-CEA antibody was administered.
  • the tumor size was measured when the antibody or IgG was administered twice a week for about one month, and the effects of ⁇ -TCP and the antibody on the growth of the tumor were observed.
  • the outline of the experiment is shown in FIG. 12, and the result is shown in FIG.
  • the control, anti-CEA antibody, and ⁇ -TCP alone (groups 1 to 3) alone did not show the effect of suppressing tumor growth, but the group 4 combined with ⁇ -TCP and anti-CEA antibody had the effect of suppressing tumor growth. Observed.
  • Tumor size formed in nude mice 7 days after transplanting 5 ⁇ 10 6 human lung cancer cells A549 (purchased from ATCC) into the skin of 6-week-old male nude mice (BALB / c-nu / nu) was measured to calculate the tumor volume.
  • Group 2 Erbitax; anti-EGFR antibody (mouse anti-human EGFR antibody)
  • Group 3 ⁇ -TCP transplantation group
  • a disk-shaped tablet of ⁇ -TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor.
  • a plastic carrier material hereinafter referred to as a “plastic piece” of the same size, which does not contain a heterologous protein that may be immunogenic, is subcutaneously transplanted and controlled. It was. The day when ⁇ -TCP or a plastic piece was implanted was set to Day 0, and the antibody was administered intraperitoneally three times, Day 0, Day 3, and Day 7.
  • Anti-EGFR antibody was administered at a dose of 100 ⁇ g, and mouse IgG 100 ⁇ g was administered as a control in groups 1 and 2 where no anti-EGFR antibody was administered. And after setting an experimental group, the tumor diameter in Day10 was measured and the tumor volume was computed from the above-mentioned calculation formula. The experimental results are shown in FIG. The control, anti-EGFR antibody, and ⁇ -TCP alone (groups 1 to 3) alone did not show the effect of suppressing tumor growth, but the group 4 combined with ⁇ -TCP and anti-EGFR antibody had the effect of suppressing tumor growth. Observed.
  • a disk-shaped tablet of ⁇ -TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor.
  • the day on which ⁇ -TCP was transplanted was set as Day 0, and the antibody was administered intraperitoneally three times in total, Day 3, Day 9, and Day 17.
  • the dose per anti-HER2 antibody was 10 mg / kg ⁇ body weight.
  • the tumor diameter in Day14, Day21, and Day24 was measured, and the tumor volume was computed from the above-mentioned formula. The experimental results are shown in FIG.
  • the present invention can be suitably used in medical fields such as antibody therapy, adjuvant therapy of antibody therapy, and tumor therapy.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Cell Biology (AREA)
  • Inorganic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Dermatology (AREA)
  • Neurosurgery (AREA)
  • Virology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Preparation (AREA)
  • Materials For Medical Uses (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

In recent years, antibody drugs have garnered interest as a cancer treatment, but while in some cases the activity of the antibodies was too weak to achieve sufficient treatment results, in other cases, high dosage amounts led to too high a cost burden on patients. For that reason, increasing immune activity has become an important issue for antibody therapy drugs, and there is need for an antibody treatment adjuvant capable of enhancing the effect of antibody drugs and thereby suppressing the amount thereof used. If lymphocytes can be made to migrate and collect in and around a tumor, the effectiveness of an antibody drug may be further improved. The purpose of the present invention is to provide a safe and low-cost antibody treatment adjuvant therapy drug capable of enhancing the effect of antibody drugs. Implantation of a dense body of β-tricalcium phosphate (TCP) into the body functions to allow activity, migration and accumulation of immune cells. The inventors discovered that the therapeutic effect of antibody drugs can be enhanced by utilizing this function, which led to the perfection of the present invention.

Description

抗体療法の効果増強剤Antibody therapy effect enhancer
　本発明は、抗体療法の効果増強剤（抗体療法補助療法剤）や抗体療法キット、より詳しくは、対象の体内に移植されて用いられるβ－リン酸三カルシウム（ＴＣＰ：Tricalcium phosphate）を有効成分とする、抗体医薬組成物を用いる抗体療法の効果増強剤や、β－リン酸三カルシウムを含む組成物と抗体医薬組成物とからなる抗体療法キットに関する。 The present invention relates to an antibody therapy effect enhancer (antibody therapy adjuvant) or antibody therapy kit, more specifically, β-tricalcium phosphate (TCP) used as an active ingredient after transplanted into the body of a subject. And an antibody therapy kit comprising an antibody pharmaceutical composition comprising a composition containing β-tricalcium phosphate and an antibody pharmaceutical composition.
　従来の化学療法による治療は、疾患にかかった異常な細胞と正常な細胞とに同等に影響を与え、それにより時に強い副作用が起こることが問題であった。そこで、異常な細胞のみに効果を発揮する、分子標的薬剤の開発が進められている。分子標的薬剤の中でも抗体医薬を用いた抗体療法は、正常細胞に発現していない、または発現していても非常に発現量が少ない抗原に対する抗体を用いることで、極力正常細胞に影響を与えず副作用の少ない、異常な細胞に選択性が高い治療効果が期待されている。 The conventional treatment with chemotherapy has the problem that the abnormal cells affected by the disease and the normal cells are equally affected, thereby causing strong side effects. Therefore, development of molecular target drugs that exert effects only on abnormal cells has been underway. Among the molecular target drugs, antibody therapy using antibody drugs does not affect normal cells as much as possible by using an antibody against an antigen that is not expressed in normal cells or is expressed in very small amounts. A highly selective therapeutic effect is expected for abnormal cells with few side effects.
　例えばがん細胞は、正常細胞の増殖、分化、細胞死に重要な役割を果たす遺伝子の欠失、異常などにより、癌の生物学的特性ともいえる細胞増殖の促進、アポトーシス抑制、血管新生、転移などを示す。これらに関わる遺伝子ががん遺伝子やがん抑制遺伝子として同定されてきており、例えば、がん遺伝子の機能を抑制することにより、がんを治療することができると考えられている。すなわち、がんの原因となっている増殖因子や増殖因子受容体を標的とした抗体を投与することにより、正常細胞に及ぼす影響が少なく、がん細胞の増殖シグナルを抑制し、増殖を抑制または停止させることができると考えられている。 Cancer cells, for example, promote cell proliferation, which can be said to be a biological characteristic of cancer, suppression of apoptosis, angiogenesis, metastasis, etc. due to deletion or abnormality of genes that play an important role in normal cell growth, differentiation, and cell death Indicates. Genes related to these have been identified as oncogenes and tumor suppressor genes. For example, it is considered that cancer can be treated by suppressing the functions of oncogenes. In other words, administration of antibodies targeting growth factors and growth factor receptors that cause cancer has little effect on normal cells, suppresses the growth signal of cancer cells and suppresses growth or It is believed that it can be stopped.
　そして、様々ながん細胞において過剰発現が観察されている上皮増殖因子受容体２型（ＨＥＲ２；Human Epidermal Growth Factor Receptor Type2；別名ｃ－ｅｒｂＢ－２）に対するモノクローナル抗体は、抗悪性腫瘍剤ハーセプチン（HERCEPTIN、登録商標；ロシュ社製）として、同様に様々な癌細胞において過剰発現が観察されている上皮細胞増殖因子受容体（ＥＧＦＲ；Epidermal Growthfactor Receptor）に対するモノクローナル抗体は、抗悪性腫瘍剤アービタックス（ERBITUX、登録商標；メルクセローノ株式会社製）として臨床において既に使用されている。 A monoclonal antibody against epidermal growth factor receptor type 2 (HER2; Human Epidermal Growth Factor Receptor Type2; also known as c-erbB-2), whose overexpression is observed in various cancer cells, is an anti-neoplastic agent Herceptin ( HERCEPTIN (registered trademark; manufactured by Roche), a monoclonal antibody against epidermal growth factor receptor (EGFR), which is also observed to be overexpressed in various cancer cells, is an anti-neoplastic agent ERBITUX (ERBITUX). , Registered trademark; manufactured by Merck Serono Co., Ltd.).
　しかしながら、抗体の活性が弱いために治療効果が十分得られないケースや、投与量が多くなることで患者のコスト負担が高くなるケース、あるいは、抗体医薬は高価であるため、抗体医薬の使用量が制限されるケースなどがある。そのため、免疫活性の増強が抗体治療薬の重要な課題になっており、抗体医薬の効果を増強したり、それにより使用量を抑えることができる、抗体療法の補助剤が求められている。 However, there are cases where the therapeutic effect cannot be sufficiently obtained due to weak antibody activity, cases where the burden of the patient becomes high due to increased dose, or the amount of antibody drug used is high because the antibody drug is expensive. There are cases where this is restricted. Therefore, enhancement of immune activity has become an important issue for antibody therapeutics, and there is a demand for adjuvants for antibody therapy that can enhance the effects of antibody drugs and thereby reduce the amount used.
　他方、β－リン酸三カルシウムを含むリン酸カルシウム系化合物は、その優れた生体親和性により人工骨、人工歯根などの生体材料として応用研究が盛んに行われている（特許文献１，２）。本発明者らは気孔率７５％、粒径２５～７５μｍ又は７５～１０５μｍのβ－リン酸三カルシウムをシートにコーティングした、マクロファージの活動を活発化させる癌細胞抑制剤（特許文献３、非特許文献１）を開発している。しかしながら、β－リン酸三カルシウムと抗体医薬を併用する効果については全く知られていなかった。 On the other hand, calcium phosphate compounds containing β-tricalcium phosphate have been actively studied as biomaterials such as artificial bones and artificial roots due to their excellent biocompatibility (Patent Documents 1 and 2). The present inventors have coated a sheet with β-tricalcium phosphate having a porosity of 75% and a particle size of 25 to 75 μm or 75 to 105 μm, and a cancer cell inhibitor that activates macrophage activity (Patent Document 3, non-patent document) Reference 1) is being developed. However, the effect of combining β-tricalcium phosphate and an antibody drug has not been known at all.
　その他、これまでに人工的にリンパ節を構築する試みは幾つかあったが、自然のリンパ節と非常に類似した構造を持ち、しかも実際に生体内で強力な免疫機能を発揮する免疫装置の開発の成功は全くなかった。例えば、特許文献４～６には、コラーゲンスポンジを用いた支持体に支持細胞（ストローマ細胞）を播き、かかる支持体を腎臓の被膜の下に埋め込むことで、人工リンパ節となり、抗体産生、がん細胞の排除などの免疫反応を強力に起こすことが開示されている。しかしこれらはいずれも細胞を使用しており、試験管の細胞培養や、人工材料を用いた構築などの工程が必要であった。 In addition, there have been several attempts to artificially construct lymph nodes, but an immune system that has a structure very similar to that of natural lymph nodes and actually exerts a strong immune function in vivo. There was no success in development. For example, in Patent Documents 4 to 6, a support cell (stromal cell) is seeded on a support using a collagen sponge, and the support is embedded under the kidney capsule, thereby forming an artificial lymph node and producing an antibody. It is disclosed to cause an immune response such as elimination of cancer cells. However, these all use cells, and steps such as cell culture in test tubes and construction using artificial materials are necessary.
特開２０１０－２３３９１４号公報JP 2010-233914 A 特開平７－１１６１７５号公報Japanese Patent Laid-Open No. 7-116175 特開２０１０－１２６４３４号公報JP 2010-126434 A 特開２００６－１２９８３９号公報JP 2006-129839 A 特開２００４－２５５１１０号公報Japanese Patent Laid-Open No. 2004-255110 特開２００８－１６３０１５号公報JP 2008-163015 A
　近年、がんの治療方法のひとつとして、抗体医薬が注目を集めている。しかしながら、抗体の活性が弱いために治療効果が十分得られないケースや、投与量が多くなることで患者のコスト負担が高くなるケース、あるいは、抗体医薬は高価であるため、抗体医薬の使用量が制限されるケースなどがある。そのため、免疫活性の増強が抗体治療薬の重要な課題になっており、自然免疫系をターゲットし、抗体医薬の効果を増強したり、それにより使用量を抑えることができる、抗体療法の補助剤が求められている。本発明の課題は、抗体医薬の効果を増強することができる安価で安全な抗体療法補助療法剤を提供することにある。 In recent years, antibody drugs have attracted attention as one of cancer treatment methods. However, there are cases where the therapeutic effect cannot be sufficiently obtained due to weak antibody activity, cases where the burden of the patient becomes high due to increased dose, or the amount of antibody drug used is high because the antibody drug is expensive. There are cases where this is restricted. Therefore, enhancement of immune activity has become an important issue for antibody therapeutics, and it is an adjuvant for antibody therapy that can target the innate immune system and enhance the effects of antibody drugs, thereby reducing the amount used. Is required. An object of the present invention is to provide an inexpensive and safe antibody therapy adjuvant therapeutic agent that can enhance the effect of an antibody drug.
　発明者らは、病変部位近傍に移植されたβ－リン酸三カルシウム緻密体（ここでは気孔率５０％以下のものを指す）により活性・誘導・集積されたＴ細胞、Ｂ細胞、ＮＫ細胞などのリンパ球や樹状細胞が高度に組織化された三次元構造をとることにより、リンパ球が抗原及び抗原提示細胞と相互作用して抗原特異的な免疫反応（適応免疫）を開始し、抗体医薬によるがん治療効果を増強できることを見出し、本発明を完成するに至った。 The inventors have disclosed T cells, B cells, NK cells, etc. that are activated, induced, and accumulated by β-tricalcium phosphate dense bodies (here, those having a porosity of 50% or less) transplanted near the lesion site. Lymphocytes and dendritic cells have a highly organized three-dimensional structure that allows lymphocytes to interact with antigens and antigen-presenting cells to initiate antigen-specific immune responses (adaptive immunity) The present inventors have found that a cancer treatment effect by a medicine can be enhanced and completed the present invention.
　すなわち、本発明は［１］対象の体内に移植されて用いられるβ－リン酸三カルシウムを有効成分とする組成物と、抗体医薬組成物とを備えた抗体療法キットや、［２］体内が、皮下であることを特徴とする［１］記載の抗体療法キットや、［３］β－リン酸三カルシウムが病変近傍に移植されることを特徴とする［１］又は［２］記載の抗体療法キットや、［４］病変近傍が、病変から０．１～１５ｃｍの距離にある領域であることを特徴とする［３］記載の抗体療法キットや、［５］抗体医薬が、がん特異的抗体医薬であることを特徴とする［１］～［４］のいずれか記載の抗体療法キットや、［６］がん特異的抗体が、抗ＣＥＡ抗体、抗ＨＥＲ２抗体、及び抗ＥＧＦＲ抗体から選ばれる１種又は２種以上の抗体であることを特徴とする［５］記載の抗体療法キットに関する。 That is, the present invention relates to [1] an antibody therapy kit comprising a composition containing β-tricalcium phosphate as an active ingredient transplanted and used in a subject's body, and an antibody pharmaceutical composition; [2] The antibody therapy kit according to [1], characterized by being subcutaneous, and the antibody according to [1] or [2], wherein [3] β-tricalcium phosphate is transplanted in the vicinity of a lesion A therapeutic kit, or [4] an antibody therapy kit according to [3], wherein the vicinity of the lesion is a region at a distance of 0.1 to 15 cm from the lesion; The antibody therapy kit according to any one of [1] to [4], wherein [6] the cancer-specific antibody is an anti-CEA antibody, an anti-HER2 antibody, and an anti-EGFR antibody, It is one type or two or more types of antibodies selected [5] The antibody therapy kit according to [5].
　また、本発明は［７］β－リン酸三カルシウムが、気孔率５０％以下の緻密体であることを特徴とする［１］～［６］のいずれか記載の抗体療法キットや、［８］β－リン酸三カルシウムが、Ｔ細胞、Ｂ細胞、ＮＫ細胞、樹状細胞、及びマクロファージを活性化、誘導、又は集積することを特徴とする［７］記載の抗体療法キットや、［９］β－リン酸三カルシウムが、錠剤又は柱状体であることを特徴とする［１］～［８］のいずれか記載の抗体療法キットや、［１０］β－リン酸三カルシウムが、粒径０．０５～２５μｍの粒子又は顆粒であることを特徴とする［１］～［８］のいずれか記載の抗体療法キットに関する。 [7] The antibody therapy kit according to any one of [1] to [6], wherein [7] β-tricalcium phosphate is a dense body having a porosity of 50% or less; Β-tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells, and macrophages, [9] the antibody therapy kit according to [7], The β-tricalcium phosphate is a tablet or a columnar body, and the antibody therapy kit according to any one of [1] to [8], or [10] β-tricalcium phosphate has a particle size The antibody therapy kit according to any one of [1] to [8], wherein the antibody therapy kit is 0.05 to 25 μm particles or granules.
　また、本発明は［１１］対象の体内に移植されて用いられるβ－リン酸三カルシウムを有効成分とする、抗体医薬組成物を用いる抗体療法の効果増強剤や、［１２］体内が、皮下であることを特徴とする［１１］記載の抗体療法の効果増強剤や、［１３］β－リン酸三カルシウムが病変近傍に移植されることを特徴とする［１１］又は［１２］記載の抗体療法の効果増強剤や、［１４］病変近傍が、病変から０．１～１５ｃｍの距離にある領域であることを特徴とする［１３］記載の抗体療法の効果増強剤や、［１５］抗体医薬が、がん特異的抗体医薬であることを特徴とする［１１］～［１４］のいずれか記載の抗体療法の効果増強剤や、［１６］がん特異的抗体が、抗ＣＥＡ抗体、抗ＨＥＲ２抗体、及び抗ＥＧＦＲ抗体から選ばれる１種又は２種以上の抗体であることを特徴とする［１５］記載の抗体療法の効果増強剤に関する。 The present invention also provides [11] an agent for enhancing the effectiveness of antibody therapy using an antibody pharmaceutical composition comprising β-tricalcium phosphate transplanted and used in a subject's body as an active ingredient, [11] or [12], wherein the antibody therapy effect-enhancing agent described in [11] or [13] β-tricalcium phosphate is transplanted in the vicinity of a lesion Antibody therapeutic effect enhancer, [14] The antibody therapeutic effect enhancer according to [13], wherein the vicinity of the lesion is a region at a distance of 0.1 to 15 cm from the lesion, [15] The antibody therapeutic effect enhancer according to any one of [11] to [14], wherein the antibody drug is a cancer-specific antibody drug, or [16] the cancer-specific antibody is an anti-CEA antibody , One selected from anti-HER2 antibody and anti-EGFR antibody Alternatively, the antibody therapy effect-enhancing agent of [15], wherein the antibody therapy effect-enhancing agent is characterized by two or more antibodies.
　また、本発明は［１７］β－リン酸三カルシウムが、気孔率５０％以下の緻密体であることを特徴とする［１１］～［１６］のいずれか記載の抗体療法の効果増強剤や、［１８］β－リン酸三カルシウムが、Ｔ細胞、Ｂ細胞、ＮＫ細胞、樹状細胞及びマクロファージを活性化、誘導、又は集積することを特徴とする［１７］記載の抗体療法の効果増強剤や、［１９］β－リン酸三カルシウムが、錠剤又は柱状体であることを特徴とする［１１］～［１８］のいずれか記載の抗体療法の効果増強剤や、［２０］β－リン酸三カルシウムが、粒径０．０５～２５μｍの粒子または顆粒であることを特徴とする［１１］～［１８］のいずれか記載の抗体療法の効果増強剤に関する。 [17] The antibody therapy effect-enhancing agent according to any one of [11] to [16], wherein [17] β-tricalcium phosphate is a dense body having a porosity of 50% or less. , [18] β-tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells and macrophages, and enhances the effect of antibody therapy according to [17] Or [19] β-tricalcium phosphate is a tablet or a columnar body, or the antibody therapy effect-enhancing agent according to any one of [11] to [18], or [20] β- The antibody therapeutic effect enhancer according to any one of [11] to [18], wherein the tricalcium phosphate is a particle or granule having a particle size of 0.05 to 25 μm.
　本発明によれば、対象の体内に移植されて用いられるβ－リン酸三カルシウムを有効成分とする抗体療法の効果増強剤を提供することができ、抗体医薬の効果を増強することができる安価で安全な抗体療法補助療法剤により疾患を治療することができる。 According to the present invention, it is possible to provide an antibody therapeutic effect enhancer comprising β-tricalcium phosphate used as an active ingredient transplanted in the body of a subject, and to reduce the effect of an antibody drug. Diseases can be treated with safe and safe antibody therapy adjuvants.
免疫細胞とβ－ＴＣＰ顆粒（サイズ２５～７５μｍ）の結合を走査型電子顕微鏡にて観察した結果を示す。The results of observing the binding of immune cells and β-TCP granules (size 25-75 μm) with a scanning electron microscope are shown. β－ＴＣＰ緻密体（緻密体錠剤：φ５×２ｍｍ）を、走査型電子顕微鏡にて観察した結果を示す。直径２μｍの粒子は、マーカーとして用いられている。The results of observation of a dense β-TCP (dense tablet: φ5 × 2 mm) with a scanning electron microscope are shown. Particles with a diameter of 2 μm are used as markers. β－ＴＣＰ緻密体（緻密体棒剤：φ０．９×５ｍｍ）を、走査型電子顕微鏡にて観察した結果を示す。The result of observing a β-TCP dense body (dense rod: φ0.9 × 5 mm) with a scanning electron microscope is shown. β－ＴＣＰ緻密体（錠剤）を正常マウスの皮下に移植して２週間経過後、組織サンプルをＨＥ染色し、顕微鏡にて観察した。リンパ球が集積し、天然のリンパ節と似た構造となっていることを示す図である。β-TCP dense bodies (tablets) were transplanted subcutaneously into normal mice, and after 2 weeks, tissue samples were stained with HE and observed with a microscope. It is a figure which shows that a lymphocyte accumulates and it has a structure similar to a natural lymph node. β－ＴＣＰ緻密体（顆粒）サイズ０．０５～１０５μｍを正常マウスの皮下に移植して２週間経過後、組織サンプルをＨＥ染色し、顕微鏡にて観察した。サイズ７５μｍ以上のβ－ＴＣＰ顆粒はマクロファージを誘導し（Ｎ＝１３）、７５μｍ以下の顆粒はリンパ球を集め（Ｎ＝１１）、サイズ０．０５～２５μｍの微小顆粒はリンパ球集塊を形成すること（図中矢印）を示す図であるA β-TCP dense body (granule) size of 0.05 to 105 μm was transplanted subcutaneously into normal mice, and after 2 weeks, the tissue sample was stained with HE and observed with a microscope. Β-TCP granules of size 75 μm or more induce macrophages (N = 13), granules of 75 μm or less collect lymphocytes (N = 11), and microgranules of size 0.05 to 25 μm form lymphocyte aggregates It is a figure showing what to do (arrow in the figure) 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗ＣＤ４５Ｒ抗体を用いてＢ細胞を免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体によりＢ細胞が誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-like β-TCP dense tablet φ5 × 2 mm was immunohistologically stained for B cells with an anti-CD45R antibody and observed with a microscope. It is a figure which shows that a B cell is induced | guided | derived by (beta) -TCP dense body (left: weak extension x4, right: strong extension x40 arrow part). 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗ＣＤ３抗体を用いてＴ細胞を免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体によりＴ細胞が誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-shaped β-TCP dense tablet φ5 × 2 mm was immunohistochemically stained with T cells using an anti-CD3 antibody and observed with a microscope. It is a figure which shows that a T cell is induced | guided | derived by (beta) -TCP dense body (left: weak expansion * 4, right: strong expansion * 40 square arrow part). 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗ＣＤ４９ｂ抗体を用いてＮＫ細胞を免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体によりＮＫ細胞が誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-like β-TCP compact tablet φ5 × 2 mm was stained with NK cells using an anti-CD49b antibody and observed with a microscope. It is a figure which shows that a NK cell is induced | guided | derived by the beta-TCP dense body (left: weak expansion * 4, right: strong expansion * 40 square arrow part). 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗ＣＤ１１ｃ抗体を用いて樹状細胞を免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体により樹状細胞が誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-like β-TCP dense tablet φ5 × 2 mm was subjected to immunohistochemical staining of dendritic cells using an anti-CD11c antibody and observed with a microscope. It is a figure which shows that a dendritic cell is induced | guided | derived by (beta) -TCP dense body (left: weak expansion * 4, right: strong expansion * 40 square arrow part). 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗Ｆ４／８０Ｒ抗体を用いてマクロファージを免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体によりマクロファージが誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-shaped β-TCP dense tablet φ5 × 2 mm was subjected to immunohistochemical staining of macrophages using an anti-F4 / 80R antibody and observed with a microscope. It is a figure which shows that a macrophage is induced | guided | derived by (beta) -TCP dense body (left: weak expansion * 4, right: strong expansion * 40 square arrow part). 円板状のβ－ＴＣＰ緻密体錠剤φ５×２ｍｍを皮下に移植した正常マウスの組織サンプルを、抗Ｄ２－４０抗体を用いてリンパ管内皮細胞を免疫組織染色し、顕微鏡にて観察した。β－ＴＣＰ緻密体によりリンパ管形成が誘導されることを示す図である（左：弱拡 ×４、右：強拡 ×４０ 矢印部）。A tissue sample of a normal mouse transplanted subcutaneously with a disk-like β-TCP dense tablet φ5 × 2 mm was immunohistologically stained for lymphatic endothelial cells using an anti-D2-40 antibody and observed with a microscope. It is a figure which shows that lymphatic vessel formation is induced | guided | derived by the beta-TCP dense body (left: weak expansion * 4, right: strong expansion * 40 square arrow part). 円板状のβ－ＴＣＰ緻密体錠剤又は同サイズのプラスティック片をヌードマウスの皮下へ移植し、それと同時に抗ＣＥＡ抗体又はＩｇＧの腹腔内投与を開始し、週２回、計８回抗体又はＩｇＧを投与し、以降週２回腫瘍体積を測定した実験の概略を示す図である。A disk-shaped β-TCP dense tablet or a plastic piece of the same size is transplanted subcutaneously into nude mice, and simultaneously, intraperitoneal administration of anti-CEA antibody or IgG is started, and the antibody or IgG is administered twice a week for a total of 8 times. Is a diagram showing an outline of an experiment in which a tumor volume was measured twice a week thereafter. 円板状のβ－ＴＣＰ緻密体錠剤又は同サイズのプラスティック片をヌードマウスの皮下に移植した日をＤａｙ０とし、Ｄａｙ０から抗ＣＥＡ抗体（ａ－ＣＥＡ）又はＩｇＧを週２回、計８回腹腔内に投与し、以降週２回腫瘍体積を測定した結果を示す。＊は、Ｄａｙ１８において抗ＣＥＡ抗体及びβ－ＴＣＰの群（第４群）の腫瘍体積が、プラスティック片及びＩｇＧのコントロール群（第１群）、又は、プラスティック片及び抗ＣＥＡ抗体の群（第３群）それぞれに対し、Tukey's testにより有意に小さいことを表す。また、＊＊は、Ｄａｙ２１において抗ＣＥＡ抗体及びβ－ＴＣＰの群（第４群）の腫瘍体積が、プラスティック片及びＩｇＧのコントロール群（第１群）に対し、Tukey's testにより有意に小さいことを表す。ＣＴＲＬはコントロールのＩｇＧ投与を、ａ－ＣＥＡは抗ＣＥＡ抗体を示す。The day when the disc-shaped β-TCP dense tablet or plastic piece of the same size was transplanted subcutaneously into nude mice was defined as Day 0. From Day 0, anti-CEA antibody (a-CEA) or IgG was administered twice a week for a total of 8 times peritoneal cavity. The results of measuring the tumor volume twice a week thereafter are shown. * In Day 18, the tumor volume of the group of anti-CEA antibody and β-TCP (group 4) is the plastic strip and IgG control group (group 1), or the group of plastic strip and anti-CEA antibody (group 3). Each group) is significantly smaller by Tukey's test. In addition, ** indicates that the tumor volume of the anti-CEA antibody and β-TCP group (Group 4) in Day 21 is significantly smaller than that of the plastic strip and IgG control group (Group 1) by Tukey's test. To express. CTRL indicates control IgG administration, and a-CEA indicates anti-CEA antibody. 円板状のβ－ＴＣＰ緻密体錠剤又は同サイズのプラスティック片をヌードマウスの皮下に移植した日をＤａｙ０とし、Ｄａｙ０から抗ＥＧＦＲ抗体（Erbitax）又はＩｇＧを、計３回腹腔内に投与し、腫瘍体積を測定した結果を示す。＊は、Ｄａｙ１０において抗ＥＧＦＲ抗体及びβ－ＴＣＰの群（第４群）の腫瘍体積が、プラスティック片及びＩｇＧのコントロール群（第１群）対し、Tukey's testにより有意に小さいことを表す。The day when the disc-like β-TCP dense tablet or plastic piece of the same size was transplanted subcutaneously into nude mice was defined as Day 0, and anti-EGFR antibody (Erbitax) or IgG was administered intraperitoneally three times from Day 0. The result of measuring the tumor volume is shown. * Indicates that the tumor volume of the anti-EGFR antibody and β-TCP group (Group 4) at Day 10 is significantly smaller by Tukey's test compared to the plastic strip and IgG control group (Group 1). 円板状のβ－ＴＣＰ緻密体錠剤ヌードマウスの皮下に移植した日をＤａｙ０とし、抗ＨＥＲ２抗体（ハーセプチン）をＤａｙ３、９、１７の計３回腹腔内に投与し、腫瘍体積をＤａｙ１４、２１、２４に測定した結果を示す。＊は、各測定日において抗ＨＥＲ２抗体及びβ－ＴＣＰの群（第４群）の腫瘍体積が、コントロール群（第１群）対し、Tukey's testにより有意に小さいことを表す。The day of subcutaneous implantation of a disc-shaped β-TCP dense tablet nude mouse was set as Day 0, and anti-HER2 antibody (Herceptin) was administered intraperitoneally three times (Day 3, 9, 17), and the tumor volume was set to Day 14, 21. , 24 shows the measurement results. * Indicates that the tumor volume of the anti-HER2 antibody and β-TCP group (Group 4) on each measurement day is significantly smaller than that of the control group (Group 1) by Tukey's test.
　本発明の抗体療法キットとしては、体内に移植されて用いられる、換言すれば体内に移植するためのβ－リン酸三カルシウム（以下、「β－ＴＣＰ」という）を有効成分とする組成物と、抗体医薬組成物とを備えたものであれば特に制限されず、また本発明の抗体医薬組成物を用いる抗体療法の効果増強剤としては、β－ＴＣＰを有効成分とする組成物であれば特に制限されず、本発明の一形態として、対象の体内にβ－リン酸三カルシウムを有効成分とする組成物を移植した後に、抗体医薬組成物を投与する抗体療法や、抗体医薬組成物を用いる抗体療法の効果増強剤製造のための、対象の体内に移植されて用いられるβ－リン酸三カルシウムの使用を挙げることができる。上記β－ＴＣＰはＣａ３（ＰＯ４）２で表される組成物の一結晶形態であって、常温において安定であり生体親和性のある化合物である。β－ＴＣＰとしては、市販のβ－ＴＣＰ粉末やβ－ＴＣＰブロック、例えば人工骨として利用されるβ－ＴＣＰブロックなどを用いてもよいし、公知の方法（特開２００４－２６６４８など）により製造してもよく、またα－ＴＣＰ粉末から製造する（特開２００６－８９３１１など）こともできる。また、本発明のβ－ＴＣＰ組成物は、実質的にβ－ＴＣＰ組成からなるものや、界面活性剤や水などを混入して製造されたものでもよいが、β－ＴＣＰブロックを粒子又は顆粒状に加工したβ－ＴＣＰの粉末の他、β－ＴＣＰ粉末を打錠により緻密化、あるいは焼結により脱泡の上緻密化した、高強度、高密度、低気孔率である、多結晶体のβ－ＴＣＰ緻密体、中でも気孔率５０％以下のβ－ＴＣＰ緻密体であることが好ましい。本発明において、β－ＴＣＰ緻密体とは気孔率５０％以下のβ－ＴＣＰをいう。また本発明において、粉末、粒子、顆粒はいずれも粒状の形態を指し、実質的な区別はない。 The antibody therapy kit of the present invention comprises a composition comprising β-tricalcium phosphate (hereinafter referred to as “β-TCP”) as an active ingredient, which is used after being implanted in the body, in other words, for implantation in the body. The antibody therapeutic composition is not particularly limited as long as it comprises an antibody pharmaceutical composition, and the antibody therapy effect-enhancing agent using the antibody pharmaceutical composition of the present invention is a composition containing β-TCP as an active ingredient. There is no particular limitation, and as one form of the present invention, there is provided antibody therapy in which an antibody pharmaceutical composition is administered after transplanting a composition containing β-tricalcium phosphate as an active ingredient into the body of a subject, or an antibody pharmaceutical composition. The use of β-tricalcium phosphate that is transplanted and used in the body of a subject for the production of an antibody therapy effect enhancer to be used can be mentioned. Β-TCP is a single crystal form of a composition represented by Ca 3 (PO 4 ) 2 and is a compound that is stable at room temperature and has biocompatibility. As β-TCP, commercially available β-TCP powder or β-TCP block, for example, a β-TCP block used as an artificial bone, or the like may be used, or it may be produced by a known method (Japanese Patent Application Laid-Open No. 2004-26648). Alternatively, it can be produced from α-TCP powder (JP 2006-89311 A). The β-TCP composition of the present invention may be substantially composed of a β-TCP composition, or may be produced by mixing a surfactant, water, or the like. In addition to the β-TCP powder processed into a shape, the β-TCP powder is densified by tableting, or defoamed by sintering to be densified, and has a high strength, high density, and low porosity. A β-TCP dense body, particularly a β-TCP dense body having a porosity of 50% or less is preferable. In the present invention, the β-TCP dense body means β-TCP having a porosity of 50% or less. In the present invention, powder, particles, and granules all indicate granular forms, and there is no substantial distinction.
　かかるβ－ＴＣＰ緻密体を構成する多結晶体の粒子サイズとして、好ましくは０．００１～５０μｍ、より好ましくは０．０１～２０μｍ、さらに好ましくは０．７５～１０．０μｍであり、平均の多結晶体の粒子サイズは好ましくは１．８０μｍである。さらに、多結晶体の粒子の隙間は好ましくは０．００１～１０μｍ、より好ましくは０．００５～５μｍ、０．０１～１．０μｍであり、平均の多結晶体の粒子の隙間は好ましくは０．０５～０．２μｍ、より好ましくは０．１μｍである。かかる多結晶体の粒子の隙間の大きさや数により、β－ＴＣＰ緻密体の気孔が定められる。 The particle size of the polycrystalline body constituting such a β-TCP dense body is preferably 0.001 to 50 μm, more preferably 0.01 to 20 μm, still more preferably 0.75 to 10.0 μm. The grain size of the crystal is preferably 1.80 μm. Furthermore, the gap between the polycrystalline particles is preferably 0.001 to 10 μm, more preferably 0.005 to 5 μm, and 0.01 to 1.0 μm. The average polycrystalline particle gap is preferably 0. .05 to 0.2 μm, more preferably 0.1 μm. The pores of the β-TCP dense body are determined by the size and number of the gaps between the polycrystalline particles.
　β－ＴＣＰの気孔率は、細孔に水銀を浸入させるために圧力を加え、圧力と圧入された水銀量から比表面積や細孔分布を求める水銀法や、検量線を用いて密度から測定することもできる。また、気孔率が０％のβ－ＴＣＰ石（測定対象の焼結体と同じ組成、結晶形の気孔をもたないβ－ＴＣＰ焼結体）の真密度ρを測定しておき、測定対象の焼結体の体積と重さから算出した見かけ密度ρ’とから気孔率Ｐ（％）＝（１－ρ’／ρ）×１００として算出することができる。また、気孔率Ｐは、例えば、アキュピック１３３０シリーズ（株式会社島津製作所製）を用いて測定することもできる。β－ＴＣＰブロックの水銀法にて計測した気孔率は、好ましくは５０～９０％、より好ましくは６０～８５％、さらに好ましくは７０～８０％である。粒径７５～１０５μｍのβ－ＴＣＰ粉末はβ－ＴＣＰブロックを粉砕して作製することもでき、これを水銀法にて計測したそれぞれの粉末の気孔率は、好ましくは５０～９０％、より好ましくは５５～８５％、さらに好ましくは５７～８０％であり、好ましい平均気孔率は６０～７０％である。また、粒径２５μｍ以下のβ－ＴＣＰミクロン顆粒はβ－ＴＣＰブロックを粒子状に粉砕して作製することもでき、これを水銀法にて計測した気孔率は好ましくは５０％以下、より好ましくは４０％以下、さらに好ましくは３０％以下であり、好ましい平均気孔率は３．５～４．５％である。β－ＴＣＰ緻密体としては、気孔率が５０％以下であるβ－ＴＣＰ粉末、粒子、顆粒、錠剤、柱状体等を例示することができ、これらはβ－ＴＣＰのみからなるものであっても、β－ＴＣＰを有効成分とし、他の成分を含むものであってもよい。５０％以下のβ－ＴＣＰの気孔率は、４０％以下が好ましく、３０％以下がより好ましく、中でも２０％以下が特に好ましい。β－ＴＣＰ粉末を打錠により緻密化したβ－ＴＣＰ緻密体の、密度から算定した気孔率としては、好ましくは５０％以下、より好ましくは４０％以下、さらに好ましくは３０％以下、中でも２０％以下を好適に例示することができる。また、焼結により脱泡の上緻密化したβ－ＴＣＰ緻密体は、密度から算定した気孔率が、好ましくは５０％以下、より好ましくは４０％以下、さらに好ましくは３０％以下、中でも２０％以下を好適に例示することができる。 The porosity of β-TCP is measured from the density by applying a pressure to inject mercury into the pores and determining the specific surface area and pore distribution from the pressure and the amount of mercury injected, and a calibration curve. You can also. In addition, the true density ρ of β-TCP stone having a porosity of 0% (β-TCP sintered body having the same composition as the object to be measured and having no crystal-like pores) is measured, and the object to be measured From the apparent density ρ ′ calculated from the volume and weight of the sintered body, porosity P (%) = (1−ρ ′ / ρ) × 100 can be calculated. The porosity P can also be measured using, for example, AccuPick 1330 series (manufactured by Shimadzu Corporation). The porosity of the β-TCP block measured by the mercury method is preferably 50 to 90%, more preferably 60 to 85%, still more preferably 70 to 80%. Β-TCP powder having a particle size of 75 to 105 μm can also be prepared by pulverizing β-TCP block, and the porosity of each powder measured by mercury method is preferably 50 to 90%, more preferably Is 55 to 85%, more preferably 57 to 80%, and a preferred average porosity is 60 to 70%. Further, β-TCP micron granules having a particle size of 25 μm or less can be prepared by pulverizing β-TCP blocks into particles, and the porosity measured by the mercury method is preferably 50% or less, more preferably It is 40% or less, more preferably 30% or less, and a preferable average porosity is 3.5 to 4.5%. Examples of the β-TCP dense body include β-TCP powder, particles, granules, tablets, and columnar bodies having a porosity of 50% or less, and these may be composed only of β-TCP. , Β-TCP may be an active ingredient and other ingredients may be included. The porosity of β-TCP of 50% or less is preferably 40% or less, more preferably 30% or less, and particularly preferably 20% or less. The porosity calculated from the density of the β-TCP dense body obtained by densifying β-TCP powder by tableting is preferably 50% or less, more preferably 40% or less, still more preferably 30% or less, especially 20%. The following can be illustrated suitably. In addition, the β-TCP dense body defoamed by sintering has a porosity calculated from the density of preferably 50% or less, more preferably 40% or less, still more preferably 30% or less, especially 20%. The following can be illustrated suitably.
　焼結による緻密化は、好ましくは６００℃超～２０００℃、より好ましくは７５０～１５００℃、さらに好ましくは９００～１３００℃で、また好ましくは１～１００時間、より好ましくは１０～８０時間、さらに好ましくは２０～５０時間焼結する例を挙げることができる。また、焼結は複数の温度にて複数回に分けて行ってもよい。例えば、錠剤のβ－ＴＣＰ緻密体は以下の方法で製造することもできる。リン酸水素カルシウム、炭酸カルシウム及び水を適切な割合で混合してスラリーを調製し、調製したスラリーを摩砕しながら反応させた後に乾燥させる。次に、乾燥して得られた固形物を粉砕して仮焼し、β－ＴＣＰ粉末が得られる。得られたβ－ＴＣＰ粉末を圧縮形成されることにより柱状体に賦形される。圧縮形成された柱状体を６００℃～１３００℃で２０時間焼結する。 Densification by sintering is preferably more than 600 ° C. to 2000 ° C., more preferably 750 to 1500 ° C., further preferably 900 to 1300 ° C., and preferably 1 to 100 hours, more preferably 10 to 80 hours, A preferred example is sintering for 20 to 50 hours. In addition, the sintering may be performed in a plurality of times at a plurality of temperatures. For example, a β-TCP dense body of a tablet can be produced by the following method. A slurry is prepared by mixing calcium hydrogen phosphate, calcium carbonate and water in an appropriate ratio, and the prepared slurry is reacted while being ground and then dried. Next, the solid obtained by drying is pulverized and calcined to obtain β-TCP powder. The obtained β-TCP powder is compressed into a columnar body. The compressed columnar body is sintered at 600 ° C. to 1300 ° C. for 20 hours.
　上記気孔率５０％以下の緻密体であるβ－ＴＣＰ粉末や粒子、顆粒の製造方法としては、例えば、リン酸水素カルシウムと炭酸カルシウムの粉末をモル比で１：１～３、好ましくは１：１．５～２．５になるように秤量して混合し、かかるリン酸水素カルシウム－炭酸カルシウム混合物に純水を添加してスラリーを調製し、調製されたスラリーをボールミルにより約２４時間湿式磨砕処理を行うことでメカノケミカル法による反応をすすめ、磨砕処理されたスラリーを、７０～９０℃、好ましくは７５～８５℃にて乾燥し、乾燥して得られた固形物を粉砕して得られたβ－ＴＣＰ粉末を７００～８００℃にて、数時間仮焼することで、β－ＴＣＰ仮焼粉末を作製し、得られたβ－ＴＣＰ仮焼粉末をふるいにかけることで各粒径の分画を得ることができ、分画ごとの気孔率を測定することで、気孔率５０％以下の分画を得ることができる。具体的には、まずふるい目が１０５μｍのふるいを通過し、かつ、ふるい目が７５μｍのふるいにかけ残ったβ－ＴＣＰを分画（Ａ）とし、次にふるい目が７５μｍのふるいを通過し、かつ、ふるい目が２５μｍのふるいにかけ残ったβ－ＴＣＰを分画（Ｂ）とし、さらに２５μｍのふるいを通過したβ－ＴＣＰを分画（Ｃ）とした場合に、分画（Ｃ）は、気孔率が確実に５０％以下のβ－ＴＣＰとなる。 As a method for producing the above-described dense β-TCP powder, particles, and granules having a porosity of 50% or less, for example, calcium hydrogen phosphate and calcium carbonate powder are used in a molar ratio of 1: 1 to 3, preferably 1: Weigh and mix to 1.5 to 2.5, add pure water to the calcium hydrogen phosphate-calcium carbonate mixture to prepare a slurry, and wet-polish the prepared slurry with a ball mill for about 24 hours. The reaction by the mechanochemical method is promoted by pulverization treatment, the slurry subjected to the pulverization treatment is dried at 70 to 90 ° C., preferably 75 to 85 ° C., and the solid matter obtained by drying is pulverized. The obtained β-TCP powder is calcined at 700 to 800 ° C. for several hours to produce a β-TCP calcined powder, and the obtained β-TCP calcined powder is sieved to obtain each particle. To obtain a fraction of diameter Can be, by measuring the porosity of each fractionation, can be obtained porosity of 50% or less of the fractions. Specifically, first, the sieve passes through a 105 μm sieve, and the sieve passes through the 75 μm sieve to leave the remaining β-TCP as a fraction (A), and then the sieve passes through a 75 μm sieve. In addition, when β-TCP remaining through a sieve having a sieve size of 25 μm is defined as fraction (B), and β-TCP that has passed through a 25 μm sieve is defined as fraction (C), fraction (C) is Β-TCP with a porosity of 50% or less is ensured.
　上記β－ＴＣＰ錠剤やβ－ＴＣＰ柱状体の製造方法としては、気孔率が５０％以下の、単一相又はほぼ単一相のβ－ＴＣＰの錠剤や柱状体を製造することができる方法を挙げることができる。具体的には、前述のようにβ－ＴＣＰ仮焼粉末を作製し、得られたβ－ＴＣＰ仮焼粉末を圧縮成形（打錠）することにより錠剤や柱状体に賦形し、かかる賦形されたβ－ＴＣＰ仮焼錠剤や柱状体を４５０～６５０℃、好ましくは６００℃にて２．５～３．５時間、好ましくは３時間焼結し、次いで８５０～９５０℃、好ましくは９００℃で０．５～１．５時間、好ましくは１時間焼結し、さらに１０００～１３００℃で０．７５～１．５時間、好ましくは１時間焼結することで燒結体としてのβ－ＴＣＰの錠剤や柱状体を得る方法を挙げることができ、また上記β－ＴＣＰ仮焼粉末を燒結し、β－ＴＣＰの焼結体としての粉体を得た後に圧縮成形（打錠）してβ－ＴＣＰの錠剤や柱状体を作製してもよい。なお、β－ＴＣＰ柱状体は、β－ＴＣＰ錠剤を柱状に打ち抜くことにより作製してもよい。 As a method for producing the β-TCP tablet or β-TCP columnar body, there is a method capable of producing a single-phase or almost single-phase β-TCP tablet or columnar body having a porosity of 50% or less. Can be mentioned. Specifically, the β-TCP calcined powder is prepared as described above, and the obtained β-TCP calcined powder is molded into tablets or columns by compression molding (tabletting). The sintered β-TCP calcined tablet or columnar body is sintered at 450 to 650 ° C., preferably 600 ° C. for 2.5 to 3.5 hours, preferably 3 hours, and then 850 to 950 ° C., preferably 900 ° C. Is sintered for 0.5 to 1.5 hours, preferably 1 hour, and further sintered at 1000 to 1300 ° C. for 0.75 to 1.5 hours, preferably 1 hour. Examples of methods for obtaining tablets and columnar bodies can be mentioned. The β-TCP calcined powder is sintered to obtain a powder as a sintered body of β-TCP, which is then compression-molded (tablet). TCP tablets and columns may be produced. The β-TCP columnar body may be produced by punching a β-TCP tablet into a columnar shape.
　本発明のβ－ＴＣＰ組成物には、β－ＴＣＰの他に薬学的に許容される通常の担体、結合剤、安定化剤、賦形剤、希釈剤、ｐＨ緩衝剤、崩壊剤、可溶化剤、溶解補助剤、等張剤などの各種調剤用配合成分を含んでいてもよく、ゲル状やペースト状、微粒子の懸濁液や、固形状の製剤とすることができるが、好ましくは固形状であり、その形状としては、顆粒状、粉末状、粒子状の他、円板状（短円柱状）、棒状（長円柱状）の錠剤などを挙げることができる。 In addition to β-TCP, the β-TCP composition of the present invention includes conventional pharmaceutically acceptable carriers, binders, stabilizers, excipients, diluents, pH buffering agents, disintegrants, solubilizing agents. May contain various preparation compounding ingredients such as an agent, a solubilizing agent, and an isotonic agent, and may be in the form of a gel, paste, suspension of fine particles, or a solid preparation. Examples of the shape include granules, powders, particles, disks (short cylinders), rods (long cylinders), and the like.
　また、固形状のβ－ＴＣＰ組成物の粒径としては、例えば、β－ＴＣＰブロックを粒子状に加工したβ－ＴＣＰ粉末の場合は、粒径が好ましくは５０～１２０μｍ、より好ましくは６０～１１０μｍ、さらに好ましくは７５～１０５μｍである。また、β－ＴＣＰブロックを粒子状に加工したβ－ＴＣＰミクロン顆粒の場合は、粒径が好ましくは５０μｍ以下、より好ましくは４０μｍ以下、さらに好ましくは３０μｍ以下である。中でも、粒径が０．０５～５μｍのβ－ＴＣＰミクロン顆粒や、粒径が０．０５～２５μｍのβ－ＴＣＰミクロン顆粒を好適に例示することができる。本発明において粒径０．０５～２５μｍの粒子は、２５μｍのふるいを通過したものを指す。円板状又は柱状や棒状の錠剤のβ－ＴＣＰ組成物の大きさとしては、直径が好ましくは０．０１～３０ｍｍ、より好ましくは０．１～２０ｍｍ、さらに好ましくは１～１０ｍｍであり、長さが好ましくは０．１～４０ｍｍ、より好ましくは０．５～３０ｍｍ、さらに好ましくは１～２０ｍｍであるが、中でも直径５ｍｍで長さ２ｍｍの円板状の錠剤や、直径０．９ｍｍで長さ１０ｍｍの柱状の棒剤を好適に例示することができる。 The particle diameter of the solid β-TCP composition is, for example, in the case of β-TCP powder obtained by processing a β-TCP block into particles, the particle diameter is preferably 50 to 120 μm, more preferably 60 to It is 110 μm, more preferably 75 to 105 μm. In the case of β-TCP micron granules obtained by processing β-TCP blocks into particles, the particle size is preferably 50 μm or less, more preferably 40 μm or less, and even more preferably 30 μm or less. Among these, β-TCP micron granules having a particle size of 0.05 to 5 μm and β-TCP micron granules having a particle size of 0.05 to 25 μm can be preferably exemplified. In the present invention, particles having a particle size of 0.05 to 25 μm refer to particles that have passed through a 25 μm sieve. The size of the β-TCP composition of a disk-shaped or columnar or rod-shaped tablet is preferably 0.01 to 30 mm in diameter, more preferably 0.1 to 20 mm, and still more preferably 1 to 10 mm. Is preferably 0.1 to 40 mm, more preferably 0.5 to 30 mm, and still more preferably 1 to 20 mm. Among them, a tablet having a diameter of 5 mm and a length of 2 mm, or a long tablet having a diameter of 0.9 mm is preferable. A columnar bar having a thickness of 10 mm can be suitably exemplified.
　本発明における体内としては、骨や歯以外の部位であれば特に制限されず、皮下、皮内、筋肉内、腹腔内、胸腔内、脳内を挙げることができ、好ましくは皮下、さらに好ましくは治療対象である病変の近傍を挙げることができる。病変の近傍に移植して用いられる場合は、好ましくは病変から１μｍ～３０ｃｍ、より好ましくは１００μｍ～２０ｃｍ、さらに好ましくは１ｍｍ～１５ｃｍの距離にある領域に移植して用いることができる。本発明の治療対象の病変は、骨や歯以外における病変であれば特に制限はされず、がん、アレルギー免疫、感染症その他を例示することができ、中でもがんを好適に挙げることができ、頭蓋骨内組織、肺、皮膚、軟組織、膀胱、胃、すい臓、頭部、頸部、腎臓、前立腺、大腸、小腸、食道、女性器（例えば卵巣、子宮）又は甲状腺のがんを特に好適に例示することができる。 The body in the present invention is not particularly limited as long as it is a site other than bones and teeth, and can include subcutaneous, intradermal, intramuscular, intraperitoneal, intrathoracic, and intracerebral, preferably subcutaneously, more preferably The vicinity of the lesion to be treated can be cited. When used by transplanting in the vicinity of a lesion, it can be used by transplanting into a region at a distance of preferably 1 μm to 30 cm, more preferably 100 μm to 20 cm, still more preferably 1 mm to 15 cm from the lesion. The lesion to be treated according to the present invention is not particularly limited as long as it is a lesion other than a bone or a tooth, and examples thereof include cancer, allergic immunity, infectious diseases, etc. Among them, cancer can be preferably mentioned. Particularly suitable for intracranial tissue, lung, skin, soft tissue, bladder, stomach, pancreas, head, neck, kidney, prostate, large intestine, small intestine, esophagus, female genitals (eg ovary, uterus) or thyroid cancer It can be illustrated.
　本発明のβ－ＴＣＰの投与方法としては、外科的処置により体内へ直接移植する方法であれば特に制限されず、他の目的の手術の際に、切開箇所に本発明のβ－ＴＣＰを移植することもできる。また生体への負担を軽減する観点から、通常の注射器や固形物の挿入用注射器によって、最小限の開口部から本発明のβ－ＴＣＰ組成物を体内に移植する方法を好適に例示することができる。この場合、本発明のβ－ＴＣＰ組成物としては、β－ＴＣＰ顆粒を含む懸濁液や、ゲル状やペースト状に調製されたβ－ＴＣＰや固形β－ＴＣＰを充填した注射器、例えば棒状（長円柱状）のβ－ＴＣＰ緻密体が充填されたデバイス（導入装置）や、棒状（長円柱状）β－ＴＣＰ緻密体とそのデバイス（導入装置）のセットを挙げることができる。また、投与量は、疾病の種類、病変の大きさ、患者の体重等により適宜選定することができ、好ましくは０．０１ｍｇ～１００ｇ、より好ましくは０．１ｍｇ～１０ｇ、さらに好ましくは１ｍｇ～１ｇを例示することができる。 The administration method of β-TCP of the present invention is not particularly limited as long as it is a method of transplanting directly into the body by a surgical procedure, and the β-TCP of the present invention is transplanted at the incision site at the time of surgery for other purposes. You can also Further, from the viewpoint of reducing the burden on the living body, a method of transplanting the β-TCP composition of the present invention into the body from a minimum opening by using a normal syringe or a syringe for inserting a solid material can be suitably exemplified. it can. In this case, the β-TCP composition of the present invention includes a suspension containing β-TCP granules, a syringe filled with β-TCP or a solid β-TCP prepared in a gel or paste form, such as a rod-like ( Examples include a device (introduction device) filled with a β-TCP dense body having a long cylindrical shape and a set of a rod-like (long cylindrical shape) β-TCP dense body and its device (introduction device). The dose can be appropriately selected according to the type of disease, the size of the lesion, the weight of the patient, etc., preferably 0.01 mg to 100 g, more preferably 0.1 mg to 10 g, and still more preferably 1 mg to 1 g. Can be illustrated.
　本発明における抗体医薬組成物としては抗体医薬を含む組成物であれば特に制限されず、好ましくはがん細胞に特異的に発現している抗原、あるいは正常細胞では低い発現を示すが、がん細胞で高い発現を示す抗原に対する抗体を含む組成物であり、抗体として作製することも、抗体あるいは抗体医薬組成物として市販品を入手することもできる。具体的には、抗ＣＥＡ抗体、抗ＨＥＲ２抗体（トラスツズマブ：Trastuzumab、商品名ハーセプチン（登録商標）（中外製薬株式会社）)、抗ＥＧＦＲ抗体（アービタックス：ERBITUX）、抗ＣＤ２０抗体（リツキシマブ：RITUXIMABやイブリツモマブ）、抗ＣＤ５２抗体（アレツズマブ；ALEMTUZUMAB、別名CAMPATH）、抗１７－１Ａ（ヒト腫瘍関連上皮細胞接着因子）抗体（エドレコロマブ：edrecolomab）、抗ＶＥＧＦ（血管内皮細胞増殖因子）抗体（ベバシツマブ：BEVACIZUMAB、商品名アバスチン（登録商標）（ジェネンテック社））、抗ＴＮＦ－α抗体（アダリムマブ：adalimumab、商品名ヒュミラ（登録商標）（エーザイ株式会社）や、インフリキシマブ：Infliximab、商品名レミケード（登録商標）（田辺三菱製薬株式会社））や、それらを含む医薬組成物を例示することができる。かかる抗体医薬組成物は薬学的に許容される通常の担体、結合剤、安定化剤、賦形剤、希釈剤、ｐＨ緩衝剤、崩壊剤、可溶化剤、溶解補助剤、等張剤などの各種調剤用配合成分を含んでいてもよく、経口的又は非経口的に投与することができ、例えば粉末、顆粒、錠剤、カプセル剤、シロップ剤、懸濁液等の剤型で経口的に投与することができ、あるいは、例えば溶液、乳剤、懸濁液等の剤型にしたものを注射により非経口投与することができる他、スプレー剤の形で鼻孔内投与することもできるが、注射や点滴による非経口投与を好適に例示することができる。 The antibody pharmaceutical composition in the present invention is not particularly limited as long as it is a composition containing an antibody pharmaceutical, and preferably shows an antigen specifically expressed in cancer cells or low expression in normal cells. It is a composition containing an antibody against an antigen that exhibits high expression in cells, and it can be prepared as an antibody, or a commercially available product can be obtained as an antibody or antibody pharmaceutical composition. Specifically, anti-CEA antibody, anti-HER2 antibody (Trastuzumab: Trastuzumab, trade name Herceptin (registered trademark) (Chugai Pharmaceutical Co., Ltd.)), anti-EGFR antibody (Arbitux: ERBITUX), anti-CD20 antibody (rituximab: RITUXIMAB and ibritumomab ), Anti-CD52 antibody (aletuzumab; ALEMTUZUMAB, also known as CAMPATH), anti-17-1A (human tumor-associated epithelial cell adhesion factor) antibody (edrecolomab), anti-VEGF (vascular endothelial growth factor) antibody (bevacizumab: BEVACIZUMAB, product) Name Avastin (registered trademark) (Genentech), anti-TNF-α antibody (adalimumab: adalimumab, trade name Humira (registered trademark) (Eisai Co., Ltd.), infliximab: Infliximab, trade name Remicade (registered trademark) (Mitsubishi Tanabe) Pharmaceutical Co., Ltd.)) and pharmaceutical compositions containing them It is possible. Such antibody pharmaceutical compositions include pharmaceutically acceptable ordinary carriers, binders, stabilizers, excipients, diluents, pH buffers, disintegrants, solubilizers, solubilizers, isotonic agents, and the like. It may contain various compounding ingredients and can be administered orally or parenterally. For example, it is orally administered in dosage forms such as powders, granules, tablets, capsules, syrups and suspensions. Or can be administered parenterally by injection in the form of a solution, emulsion, suspension, etc., or can be administered intranasally in the form of a spray, A parenteral administration by drip can be preferably exemplified.
　本発明の抗体療法キットや本発明の抗体療法の効果増強剤は、ａ）対象の体内にβ－リン酸三カルシウムを移植するステップ；及びｂ）対象に抗体医薬組成物を投与するステップ；を備える抗体医薬による抗体療法に用いることもできる。抗体医薬組成物は、β－ＴＣＰが移植されると同時、又はその前後に対象に投与され、投与量や投与回数は抗体医薬の活性の強さ、疾病の種類、患者の体重、投与形態等により適宜選定することができ、例えば、好ましくは１日あたり０．０１～１００ｍｇ／ｋｇ（体重）、より好ましくは０．０５～７０ｍｇ／ｋｇ（体重）、さらに好ましくは０．１～５０ｍｇ／ｋｇ（体重）を挙げることができる。好ましくは本発明のβ－ＴＣＰが移植されると同時、又はその後に抗体医薬組成物を対象への投与を開始し、一定期間ごと、好ましくは週２回ごとに、計８回投与する例を挙げることができる。また、本発明における抗体医薬は、抗体医薬以外の他の薬剤、サプリメントなどの投与と併用してもよく、また手術などの外科療法と併用することもできる。 The antibody therapy kit of the present invention or the effect enhancer of the antibody therapy of the present invention comprises: a) transplanting β-tricalcium phosphate into the body of a subject; and b) administering an antibody pharmaceutical composition to the subject. It can also be used for antibody therapy with an antibody drug provided. The antibody pharmaceutical composition is administered to the subject at the same time before or after β-TCP is transplanted, and the dosage and frequency of administration are the strength of the activity of the antibody pharmaceutical, the type of disease, the weight of the patient, the dosage form, etc. For example, preferably 0.01 to 100 mg / kg (body weight) per day, more preferably 0.05 to 70 mg / kg (body weight), still more preferably 0.1 to 50 mg / kg. (Weight) can be mentioned. An example in which administration of the antibody pharmaceutical composition to the subject is preferably started at the same time or after transplantation of β-TCP of the present invention, and is administered at a fixed period, preferably every 2 weeks, for a total of 8 times. Can be mentioned. In addition, the antibody drug in the present invention may be used in combination with administration of other drugs and supplements other than the antibody drug, or may be used in combination with surgical therapy such as surgery.
　本発明の抗体医薬品を用いる抗体療法の効果増強剤は、抗体医薬組成物の投与のみによる抗体療法と、コントロール又は本発明のβ－ＴＣＰ組成物及び抗体医薬組成物の投与を併用した場合の治療効果とを比較した場合に、β－ＴＣＰ組成物を併用した方が高い治療効果が得られるという効果を有する。例えば腫瘍マーカーの値の減少や、例えば腫瘍サイズを指標として比較した場合に、コントロールや抗体療法のみに比べ、抗体療法に加えてβ－ＴＣＰを併用した方が、腫瘍サイズが縮小、あるいは増加しないことを調べることにより、本発明の効果を具体的に確認することができる。 The effect enhancer of the antibody therapy using the antibody pharmaceutical of the present invention is a treatment in which the antibody therapy only by administration of the antibody pharmaceutical composition and the control or administration of the β-TCP composition and the antibody pharmaceutical composition of the present invention are used in combination. When compared with the effect, the combined use of the β-TCP composition has the effect of obtaining a higher therapeutic effect. For example, when the tumor marker value is decreased or when the tumor size is compared as an index, the tumor size is not reduced or increased when β-TCP is used in addition to the antibody therapy, compared to the control and antibody therapy alone. By examining this, the effect of the present invention can be specifically confirmed.
　また本発明は、β－ＴＣＰを移植したマウスを、新たな抗体医薬組成物の効果の検証や、候補物質の段階では薬効の弱い抗体医薬組成物のスクリーニングにも用いることもできるという効果を有する。例えば、抗腫瘍効果の検証には多くの場合ヌードマウスが用いられるが、ヌードマウスでは抗腫瘍効果をもたらす可能性のあるエフェクター細胞がＮＫ細胞だけであるのに対し、本発明における実験系では正常なマウスを用いて、ＮＫ細胞だけでなくＴ細胞及びＢ細胞の関与についても検討できるため、よりヒトの臨床（癌患者）に近い試験を行うことができる。 In addition, the present invention has an effect that a mouse transplanted with β-TCP can also be used for verification of the effect of a new antibody pharmaceutical composition and for screening of an antibody pharmaceutical composition having a weak drug effect at the stage of a candidate substance. . For example, nude mice are often used for verification of the antitumor effect, but in nude mice, the only effector cells that may have an antitumor effect are NK cells, whereas in the experimental system of the present invention, normal Since it is possible to examine the involvement of not only NK cells but also T cells and B cells using a simple mouse, a test closer to human clinical (cancer patients) can be performed.
　以下の実施例により本発明をさらに具体的に説明するが、本発明は実施例によって限定されるものではない。 The following examples further illustrate the present invention, but the present invention is not limited to the examples.
［β－ＴＣＰの作製方法］
　β－ＴＣＰブロックは、人工骨として製品を購入することもできるが、原料のβ－ＴＣＰと水、界面活性剤、発泡剤を混ぜ、焼結することにより作製することもできる。かかるβ－ＴＣＰブロックは、気孔率７５％や気孔率６０％であった。また、β－ＴＣＰ粉末やβ－ＴＣＰ顆粒については、前記気孔率７５％のβ－ＴＣＰブロックを粉砕し、篩にかけて粒径７５～１０５μｍのβ－ＴＣＰ粉末（気孔率：６０～７０％）を得た。また、同様に篩にかけて得られた２５～７５μｍのβ－ＴＣＰ（気孔率：４０～５０％）とＰＢＳ溶液を混合後、すぐに液体上部４分の３部分の液体を採取し、１０００ｒｐｍにて３分間の遠心操作で沈殿から０．０５～２５μｍのβ－ＴＣＰミクロン顆粒（気孔率：３５～５０％）を、上清から０．０５～５μｍのミクロン顆粒を採取した（気孔率：２０～３０％）。また、リン酸水素カルシウム、炭酸カルシウム及び水を適切な割合で混合してスラリーを調製し、調製したスラリーを摩砕しながら反応させた後に乾燥させ、得られた固形物を粉砕して仮焼することでβ－ＴＣＰ粉末が得られ、その粉末を圧縮形成することによりφ５×２ｍｍ錠剤に賦形し、６００～１５００℃で３０時間焼結することで、円板状のβ－ＴＣＰ緻密体錠剤を作製した（気孔率：０．１～２０％）。また、β－ＴＣＰの原料粉末３０ｇ、界面活性剤６．０ｍｌ、水１０ｍｌとを遠心管に注入し、２５０ｒｐｍで４分間処理することで脱泡処理をし、型に注入して自然乾燥させ、離型後、１３００℃にて焼結することで、φ０．９×１０ｍｍの柱状のβ－ＴＣＰ緻密体棒剤を作製した（気孔率:１０～３０％）。なお、作製した各β－ＴＣＰの気孔率は、水銀法にて、初期圧に対応する直径約１８０μｍの細孔にまで水銀が圧入された時の試料体積に対する値として測定された。 [Production Method of β-TCP]
The β-TCP block can be purchased as an artificial bone, but can also be produced by mixing and sintering raw material β-TCP and water, a surfactant and a foaming agent. Such β-TCP block had a porosity of 75% and a porosity of 60%. For β-TCP powder and β-TCP granules, the β-TCP block having a porosity of 75% is crushed and sieved to obtain a β-TCP powder having a particle size of 75 to 105 μm (porosity: 60 to 70%). Obtained. Similarly, after mixing the 25-75 μm β-TCP (porosity: 40-50%) obtained by sieving and PBS solution, the liquid in the upper three-quarters of the liquid was immediately collected, and 1000 rpm was collected. By centrifugation for 3 minutes, 0.05-25 μm β-TCP micron granules (porosity: 35-50%) were collected from the precipitate, and 0.05-5 μm micron granules were collected from the supernatant (porosity: 20- 30%). Moreover, calcium hydrogen phosphate, calcium carbonate, and water are mixed at an appropriate ratio to prepare a slurry, the prepared slurry is reacted while being ground, dried, and the obtained solid is pulverized and calcined. Β-TCP powder is obtained, and the powder is compressed and formed into a φ5 × 2 mm tablet and sintered at 600-1500 ° C. for 30 hours, so that a disk-like β-TCP dense body is obtained. Tablets were prepared (porosity: 0.1-20%). Also, 30 g of β-TCP raw material powder, 6.0 ml of surfactant, and 10 ml of water are poured into a centrifuge tube, defoamed by being treated at 250 rpm for 4 minutes, poured into a mold, and naturally dried. After releasing the mold, sintering was performed at 1300 ° C. to prepare a columnar β-TCP dense rod having a diameter of 0.9 mm × 10 mm (porosity: 10 to 30%). The porosity of each β-TCP produced was measured by the mercury method as a value relative to the sample volume when mercury was pressed into pores having a diameter of about 180 μm corresponding to the initial pressure.
　作製したβ－ＴＣＰ顆粒（サイズ２５～７５μｍ）と免疫細胞の結合を走査型電子顕微鏡にて観察した結果を図１に示す。また、作製したβ－ＴＣＰ緻密体錠剤、及び緻密体棒剤を、走査型電子顕微鏡にて観察した結果を図２、図３に示す。また、β－ＴＣＰ緻密体を皮下に移植した正常マウスから組織サンプルを採取し、ＨＥ染色し、顕微鏡にて観察した結果を図４に示す。リンパ球が集積し、天然のリンパ節と似た構造となっていることがわかった。また、サイズ０．０５～１０５μｍのβ－ＴＣＰ顆粒（気孔率０～７０％）を皮下に移植したマウスから組織サンプルを採取し、ＨＥ染色し、顕微鏡にて観察した結果を図５に示す。サイズ７５μｍ以上のβ－ＴＣＰ顆粒はマクロファージを誘導し（Ｎ＝１３）、７５μｍ以下の顆粒はリンパ球を集め（Ｎ＝１１）、サイズ０．０５～２５μｍの微小顆粒はリンパ球集塊を形成した。また、φ５×２ｍｍのβ－ＴＣＰ緻密体（気孔率：１８％）を皮下に移植したマウスの組織サンプルを抗ＣＤ４５Ｒ抗体、抗ＣＤ３抗体、抗ＣＤ４９ｂ抗体、抗ＣＤ１１ｃ抗体、抗Ｆ４／８０Ｒ抗体、又は抗Ｄ２－４０抗体を用いて、それぞれＢ細胞、Ｔ細胞、ＮＫ細胞、樹状細胞、マクロファージ、リンパ管を免疫組織染色して顕微鏡にて観察した。結果を図６～図１１に示す。以上の結果から、皮下に移植された焼結により緻密化したβ－ＴＣＰは、リンパ球（Ｂ細胞、Ｔ細胞、ＮＫ細胞）、樹状細胞、マクロファージを誘導し、人工リンパ節として利用できることが確認された。抗腫瘍効果の検証には、多くの場合ヌードマウスが用いられる。しかしながらヌードマウスでは抗腫瘍効果をもたらす可能性のあるエフェクター細胞がＮＫ細胞だけであるのに対し、本発明のβ－ＴＣＰを移植したマウスは、Ｔ細胞及びＢ細胞の関与についても検討できるため、よりヒトの臨床（癌患者）に近い状態での試験を行うことができることが確認できた。 FIG. 1 shows the results of observation of the binding of the prepared β-TCP granules (size 25 to 75 μm) and immune cells with a scanning electron microscope. Further, the results of observation of the produced β-TCP dense tablet and the dense rod with a scanning electron microscope are shown in FIGS. In addition, FIG. 4 shows the results of tissue samples collected from normal mice transplanted subcutaneously with β-TCP dense bodies, stained with HE, and observed with a microscope. It was found that lymphocytes accumulate and have a structure similar to that of natural lymph nodes. Further, FIG. 5 shows the results of tissue samples collected from mice transplanted subcutaneously with β-TCP granules (porosity 0 to 70%) of size 0.05 to 105 μm, stained with HE, and observed with a microscope. Β-TCP granules of size 75 μm or more induce macrophages (N = 13), granules of 75 μm or less collect lymphocytes (N = 11), and microgranules of size 0.05 to 25 μm form lymphocyte aggregates did. Further, a tissue sample of a mouse transplanted subcutaneously with φ5 × 2 mm β-TCP dense body (porosity: 18%) was used as an anti-CD45R antibody, anti-CD3 antibody, anti-CD49b antibody, anti-CD11c antibody, anti-F4 / 80R antibody, Alternatively, using an anti-D2-40 antibody, B cells, T cells, NK cells, dendritic cells, macrophages, and lymphatic vessels were immunohistologically stained and observed under a microscope. The results are shown in FIGS. From the above results, β-TCP densified by sintering implanted subcutaneously can induce lymphocytes (B cells, T cells, NK cells), dendritic cells, and macrophages and can be used as an artificial lymph node. confirmed. Nude mice are often used for verification of antitumor effects. However, in nude mice, the only effector cells that may have an antitumor effect are NK cells, whereas mice transplanted with β-TCP of the present invention can also investigate the involvement of T cells and B cells. It was confirmed that the test can be conducted in a state closer to human clinical (cancer patients).
［β－ＴＣＰを用いた抗ＣＥＡ抗体による抗体療法］
　６週齢の雄性ヌードマウス（BALB/c-nu/nu）皮内に２．５×１０６個のヒト大腸癌細胞COLO 205（ＡＴＣＣ社より購入）を移植し、５～７日後、マウスに形成された腫瘍サイズを計測して腫瘍体積を算出した。腫瘍体積の算出は以下の数式によった。
　　　腫瘍体積＝（長径×短径２）／２　（ｍｍ３）
そして、各群の腫瘍体積が均等になるように群分けを行い、以下の処置群を設定した。
　　１群：対照群
　　２群：β－ＴＣＰ移植群
　　３群：抗ＣＥＡ抗体（マウス抗ヒトCarcinoembryonic Antigen抗体）投与群
　　４群：β－ＴＣＰ移植＋抗ＣＥＡ抗体投与群 [Antibody therapy with anti-CEA antibody using β-TCP]
Six-week-old male nude mice (BALB / c-nu / nu) were transplanted with 2.5 × 10 6 human colorectal cancer cells COLO 205 (purchased from ATCC) and 5-7 days later, Tumor volume was calculated by measuring the size of the formed tumor. The tumor volume was calculated according to the following formula.
Tumor volume = (major axis × minor axis 2 ) / 2 (mm 3 )
And it divided into groups so that the tumor volume of each group might become equal, and set up the following treatment groups.
Group 1: Control group Group 2: β-TCP transplantation group Group 3: Anti-CEA antibody (mouse anti-human Carcinoembryonic Antigen antibody) administration group Group 4: β-TCP transplantation + anti-CEA antibody administration group
　円板状のβ－ＴＣＰ緻密体（気孔率：０．１～１２％）の錠剤を、形成腫瘍から１５～２０ｍｍ離れた部位の皮下に移植した。β－ＴＣＰを移植しない１群と３群では、同サイズの、免疫原性をもつ可能性のある異種蛋白を含まないプラスチックキャリアマテリアル（以下「プラスチック片」と呼ぶ）を皮下に移植し、コントロールとした。β－ＴＣＰ又はプラスチック片を移植した日をＤａｙ０とし、Ｄａｙ０から週２回、の計８回、抗体を腹腔内に投与した。抗ＣＥＡ抗体は１回あたりの投与量５０μｇを、抗ＣＥＡ抗体を投与しない１群と２群ではコントロールとしてマウスＩｇＧ５０μｇを投与した。そして、実験群を設定後、約１ヶ月にわたり週２回の抗体またはＩｇＧ投与時に腫瘍サイズを計測、腫瘍の増殖に対するβ－ＴＣＰと抗体の作用を観察した。実験の概略を図１２に、結果を図１３に示す。コントロール及び抗ＣＥＡ抗体、β－ＴＣＰそれぞれ単独（１～３群）では腫瘍増殖抑制の効果は見られなかったが、β－ＴＣＰ及び抗ＣＥＡ抗体を併用した４群では、腫瘍増殖抑制の効果が観察された。 A tablet of disk-shaped β-TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor. In groups 1 and 3 where β-TCP is not transplanted, a plastic carrier material (hereinafter referred to as “plastic piece”) of the same size that does not contain a heterologous protein that may have immunogenicity is implanted subcutaneously and controlled. It was. The day on which β-TCP or plastic pieces were implanted was set to Day 0, and the antibody was administered intraperitoneally a total of 8 times, twice a week from Day 0. The anti-CEA antibody was administered at a dose of 50 μg per dose, and mouse IgG 50 μg was administered as a control in groups 1 and 2 where no anti-CEA antibody was administered. After setting the experimental group, the tumor size was measured when the antibody or IgG was administered twice a week for about one month, and the effects of β-TCP and the antibody on the growth of the tumor were observed. The outline of the experiment is shown in FIG. 12, and the result is shown in FIG. The control, anti-CEA antibody, and β-TCP alone (groups 1 to 3) alone did not show the effect of suppressing tumor growth, but the group 4 combined with β-TCP and anti-CEA antibody had the effect of suppressing tumor growth. Observed.
［β－ＴＣＰを用いた抗ＥＧＦＲ抗体による抗体療法］
　６週齢の雄性ヌードマウス（BALB/c-nu/nu）皮内に５×１０６個のヒト肺癌細胞A549（ＡＴＣＣ社より購入）を移植し、７日後、ヌードマウスに形成された腫瘍サイズを計測して腫瘍体積を算出した。腫瘍体積の算出は以下の数式によった。
　　　腫瘍体積＝（長径×短径２）／２　（ｍｍ３）
そして、各群の腫瘍体積が均等になるように群分けを行い、以下の処置群を設定した。
　　１群：対照群
　　２群：Erbitax；抗ＥＧＦＲ抗体（マウス抗ヒトＥＧＦＲ抗体）
　　３群：β－ＴＣＰ移植群
　　４群：β－ＴＣＰ移植＋抗ＥＧＦＲ抗体投与群 [Antibody therapy with anti-EGFR antibody using β-TCP]
Tumor size formed in nude mice 7 days after transplanting 5 × 10 6 human lung cancer cells A549 (purchased from ATCC) into the skin of 6-week-old male nude mice (BALB / c-nu / nu) Was measured to calculate the tumor volume. The tumor volume was calculated according to the following formula.
Tumor volume = (major axis × minor axis 2 ) / 2 (mm 3 )
And it divided into groups so that the tumor volume of each group might become equal, and set up the following treatment groups.
Group 1: Control group Group 2: Erbitax; anti-EGFR antibody (mouse anti-human EGFR antibody)
Group 3: β-TCP transplantation group Group 4: β-TCP transplantation + anti-EGFR antibody administration group
　円板状のβ－ＴＣＰ緻密体（気孔率：０．１～１２％）の錠剤を、形成腫瘍から１５～２０ｍｍ離れた部位の皮下に移植した。β－ＴＣＰを移植しない１群と２群では、同サイズの、免疫原性をもつ可能性のある異種蛋白を含まないプラスチックキャリアマテリアル（以下「プラスチック片」と呼ぶ）を皮下に移植し、コントロールとした。β－ＴＣＰ又はプラスチック片を移植した日をＤａｙ０とし、Ｄａｙ０、Ｄａｙ３、Ｄａｙ７の計３回、抗体を腹腔内に投与した。抗ＥＧＦＲ抗体は１回あたりの投与量１００μｇを、抗ＥＧＦＲ抗体を投与しない１群と２群ではコントロールとしてマウスＩｇＧ１００μｇを投与した。そして、実験群を設定後、Ｄａｙ１０における腫瘍径を計測、上述の算定式から腫瘍体積を算出した。
　実験結果を図１４に示す。コントロール及び抗ＥＧＦＲ抗体、β－ＴＣＰそれぞれ単独（１～３群）では腫瘍増殖抑制の効果は見られなかったが、β－ＴＣＰ及び抗ＥＧＦＲ抗体を併用した４群では、腫瘍増殖抑制の効果が観察された。 A disk-shaped tablet of β-TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor. In Group 1 and Group 2 where β-TCP is not transplanted, a plastic carrier material (hereinafter referred to as a “plastic piece”) of the same size, which does not contain a heterologous protein that may be immunogenic, is subcutaneously transplanted and controlled. It was. The day when β-TCP or a plastic piece was implanted was set to Day 0, and the antibody was administered intraperitoneally three times, Day 0, Day 3, and Day 7. Anti-EGFR antibody was administered at a dose of 100 μg, and mouse IgG 100 μg was administered as a control in groups 1 and 2 where no anti-EGFR antibody was administered. And after setting an experimental group, the tumor diameter in Day10 was measured and the tumor volume was computed from the above-mentioned calculation formula.
The experimental results are shown in FIG. The control, anti-EGFR antibody, and β-TCP alone (groups 1 to 3) alone did not show the effect of suppressing tumor growth, but the group 4 combined with β-TCP and anti-EGFR antibody had the effect of suppressing tumor growth. Observed.
［β－ＴＣＰを用いた抗ＨＥＲ２抗体による抗体療法］
　６週齢の雄性ヌードマウス（BALB/c-nu/nu）皮内に５×１０６個のヒト肺癌細胞Calu-3（ＡＴＣＣ社より購入）を移植し、ヌードマウスに形成された腫瘍サイズを計測して腫瘍体積を算出した。腫瘍体積の算出は以下の数式によった。
　　　腫瘍体積＝（長径×短径２）／２　（ｍｍ３）
そして、腫瘍体積が５０－１５０ｍｍ３となった時点で各群の腫瘍体積が均等になるように群分けを行い、以下の処置群を設定した。
　　１群：対照群
　　２群：β－ＴＣＰ移植群
　　３群：抗ＨＥＲ２抗体（ハーセプチン）投与群
　　４群：β－ＴＣＰ移植＋抗ＨＥＲ２抗体投与群 [Antibody therapy with anti-HER2 antibody using β-TCP]
Transplant 6 × 10 6 human lung cancer cells Calu-3 (purchased from ATCC) into the skin of 6-week-old male nude mice (BALB / c-nu / nu), and determine the tumor size formed in the nude mice. The tumor volume was calculated by measurement. The tumor volume was calculated according to the following formula.
Tumor volume = (major axis × minor axis 2 ) / 2 (mm 3 )
Then, when the tumor volume reached 50-150 mm 3 , the groups were divided so that the tumor volumes of each group were equal, and the following treatment groups were set.
Group 1: Control group Group 2: β-TCP transplantation group Group 3: Anti-HER2 antibody (Herceptin) administration group Group 4: β-TCP transplantation + anti-HER2 antibody administration group
　円板状のβ－ＴＣＰ緻密体（気孔率：０．１～１２％）の錠剤を、形成腫瘍から１５～２０ｍｍ離れた部位の皮下に移植した。β－ＴＣＰを移植した日をＤａｙ０とし、Ｄａｙ３、Ｄａｙ９、Ｄａｙ１７の計３回、抗体を腹腔内に投与した。抗ＨＥＲ２抗体は１回あたりの投与量を１０ｍｇ／ｋｇ・body weightとした。そして、実験群を設定後、Ｄａｙ１４、Ｄａｙ２１、及びＤａｙ２４における腫瘍径を計測、上述の算定式から腫瘍体積を算出した。
　実験結果を図１５に示す。コントロール及び抗ＨＥＲ２抗体、β－ＴＣＰそれぞれ単独（１～３群）では腫瘍増殖抑制の効果は見られなかったが、β－ＴＣＰと抗ＨＥＲ２抗体を併用した４群では、有意な腫瘍増殖抑制効果が観察された。 A disk-shaped tablet of β-TCP dense body (porosity: 0.1 to 12%) was implanted subcutaneously at a site 15 to 20 mm away from the formed tumor. The day on which β-TCP was transplanted was set as Day 0, and the antibody was administered intraperitoneally three times in total, Day 3, Day 9, and Day 17. The dose per anti-HER2 antibody was 10 mg / kg · body weight. And after setting an experimental group, the tumor diameter in Day14, Day21, and Day24 was measured, and the tumor volume was computed from the above-mentioned formula.
The experimental results are shown in FIG. The control, anti-HER2 antibody, and β-TCP alone (groups 1 to 3) alone did not show the effect of suppressing tumor growth, but the group 4 that combined β-TCP and anti-HER2 antibody had a significant tumor growth inhibitory effect. Was observed.
　本発明は、抗体療法、抗体療法の補助療法、腫瘍治療法などの医療分野などに好適に利用することができる。 The present invention can be suitably used in medical fields such as antibody therapy, adjuvant therapy of antibody therapy, and tumor therapy.

Claims (20)

  1. 　対象の体内に移植されて用いられるβ－リン酸三カルシウムを有効成分とする組成物と、抗体医薬組成物とを備えた抗体療法キット。 An antibody therapy kit comprising a composition comprising β-tricalcium phosphate as an active ingredient transplanted into a subject's body and an antibody pharmaceutical composition.
  2. 　体内が、皮下であることを特徴とする請求項１記載の抗体療法キット。 2. The antibody therapy kit according to claim 1, wherein the body is subcutaneous.
  3. 　β－リン酸三カルシウムが病変近傍に移植されることを特徴とする請求項１又は２記載の抗体療法キット。 The antibody therapy kit according to claim 1 or 2, wherein β-tricalcium phosphate is transplanted in the vicinity of the lesion.
  4. 　病変近傍が、病変から０．１～１５ｃｍの距離にある領域であることを特徴とする請求項３記載の抗体療法キット。 4. The antibody therapy kit according to claim 3, wherein the vicinity of the lesion is a region at a distance of 0.1 to 15 cm from the lesion.
  5. 　抗体医薬が、がん特異的抗体医薬であることを特徴とする請求項１～４のいずれか記載の抗体療法キット。 The antibody therapy kit according to any one of claims 1 to 4, wherein the antibody drug is a cancer-specific antibody drug.
  6. 　がん特異的抗体が、抗ＣＥＡ抗体、抗ＨＥＲ２抗体、及び抗ＥＧＦＲ抗体から選ばれる１種又は２種以上の抗体であることを特徴とする請求項５記載の抗体療法キット。 6. The antibody therapy kit according to claim 5, wherein the cancer-specific antibody is one or more antibodies selected from anti-CEA antibody, anti-HER2 antibody, and anti-EGFR antibody.
  7. 　β－リン酸三カルシウムが、気孔率５０％以下の緻密体であることを特徴とする請求項１～６のいずれか記載の抗体療法キット。 7. The antibody therapy kit according to claim 1, wherein β-tricalcium phosphate is a dense body having a porosity of 50% or less.
  8. 　β－リン酸三カルシウムが、Ｔ細胞、Ｂ細胞、ＮＫ細胞、樹状細胞、及びマクロファージを活性化、誘導、又は集積することを特徴とする請求項７記載の抗体療法キット。 The antibody therapy kit according to claim 7, wherein β-tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells, and macrophages.
  9. 　β－リン酸三カルシウムが、錠剤又は柱状体であることを特徴とする請求項１～８のいずれか記載の抗体療法キット。 The antibody therapy kit according to any one of claims 1 to 8, wherein β-tricalcium phosphate is a tablet or a columnar body.
  10. 　β－リン酸三カルシウムが、粒径０．０５～２５μｍの粒子又は顆粒であることを特徴とする請求項１～８のいずれか記載の抗体療法キット。 The antibody therapy kit according to any one of claims 1 to 8, wherein β-tricalcium phosphate is a particle or granule having a particle size of 0.05 to 25 µm.
  11. 　対象の体内に移植されて用いられるβ－リン酸三カルシウムを有効成分とする、抗体医薬組成物を用いる抗体療法の効果増強剤。 An agent for enhancing the effectiveness of antibody therapy using an antibody pharmaceutical composition comprising β-tricalcium phosphate used as an active ingredient transplanted in the body of a subject.
  12. 　体内が、皮下であることを特徴とする請求項１１記載の抗体療法の効果増強剤。 12. The antibody therapeutic effect enhancer according to claim 11, wherein the body is subcutaneous.
  13. 　β－リン酸三カルシウムが病変近傍に移植されることを特徴とする請求項１１又は１２記載の抗体療法の効果増強剤。 13. The antibody therapy effect-enhancing agent according to claim 11 or 12, wherein β-tricalcium phosphate is transplanted in the vicinity of the lesion.
  14. 　病変近傍が、病変から０．１～１５ｃｍの距離にある領域であることを特徴とする請求項１３記載の抗体療法の効果増強剤。 The antibody therapeutic effect enhancer according to claim 13, wherein the vicinity of the lesion is a region located at a distance of 0.1 to 15 cm from the lesion.
  15. 　抗体医薬が、がん特異的抗体医薬であることを特徴とする請求項１１～１４のいずれか記載の抗体療法の効果増強剤。 15. The antibody therapeutic effect enhancer according to claim 11, wherein the antibody drug is a cancer-specific antibody drug.
  16. 　がん特異的抗体が、抗ＣＥＡ抗体、抗ＨＥＲ２抗体、及び抗ＥＧＦＲ抗体から選ばれる１種又は２種以上の抗体であることを特徴とする請求項１５記載の抗体療法の効果増強剤。 The antibody therapeutic effect enhancer according to claim 15, wherein the cancer-specific antibody is one or more antibodies selected from anti-CEA antibody, anti-HER2 antibody, and anti-EGFR antibody.
  17. 　β－リン酸三カルシウムが、気孔率５０％以下の緻密体であることを特徴とする請求項１１～１６のいずれか記載の抗体療法の効果増強剤。 The antibody therapy effect-enhancing agent according to any one of claims 11 to 16, wherein β-tricalcium phosphate is a dense body having a porosity of 50% or less.
  18. 　β－リン酸三カルシウムが、Ｔ細胞、Ｂ細胞、ＮＫ細胞、樹状細胞、及びマクロファージを活性化、誘導、又は集積することを特徴とする請求項１７記載の抗体療法の効果増強剤。 18. The antibody therapeutic effect enhancer according to claim 17, wherein β-tricalcium phosphate activates, induces or accumulates T cells, B cells, NK cells, dendritic cells, and macrophages.
  19. 　β－リン酸三カルシウムが、錠剤又は柱状体であることを特徴とする請求項１１～１８のいずれか記載の抗体療法の効果増強剤。 19. The antibody therapeutic effect enhancer according to claim 11, wherein β-tricalcium phosphate is a tablet or a columnar body.
  20. 　β－リン酸三カルシウムが、粒径０．０５～２５μｍの粒子又は顆粒であることを特徴とする請求項１１～１８のいずれか記載の抗体療法の効果増強剤。 19. The antibody therapy effect-enhancing agent according to claim 11, wherein β-tricalcium phosphate is a particle or granule having a particle size of 0.05 to 25 μm.
PCT/JP2012/000579 2011-01-31 2012-01-30 Antibody therapy effect-enhancing drug WO2012105219A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2012555739A JP5990752B2 (en) 2011-01-31 2012-01-30 Antibody therapy effect enhancer

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2011-018839 2011-01-31
JP2011018839 2011-01-31

Publications (1)

Publication Number Publication Date
WO2012105219A1 true WO2012105219A1 (en) 2012-08-09

Family

ID=46602444

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2012/000579 WO2012105219A1 (en) 2011-01-31 2012-01-30 Antibody therapy effect-enhancing drug

Country Status (2)

Country Link
JP (1) JP5990752B2 (en)
WO (1) WO2012105219A1 (en)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007000637A (en) * 2005-06-22 2007-01-11 Heraeus Kulzer Gmbh Moldable implant material and the use
JP2007217307A (en) * 2006-02-14 2007-08-30 Meiji Univ Support for drug delivery and pharmaceutical using the same
JP2009024014A (en) * 2001-03-26 2009-02-05 Austin Research Inst Antibody against cancer
JP2009525048A (en) * 2006-02-01 2009-07-09 ザ ジョンズ ホプキンス ユニバーシティー Polypeptide-nucleic acid complex for immunological prophylaxis or immunotherapy against neoplastic disorders or infections
JP2009195739A (en) * 2004-10-14 2009-09-03 Biomimetic Therapeutics Inc Platelet-derived growth factor composition and method of use thereof
JP2010126434A (en) * 2008-11-25 2010-06-10 Olympus Corp Cancer cell inhibitor and cancer cell-inhibiting sheet
JP2010540498A (en) * 2007-09-28 2010-12-24 ウニヴェルズィテーツシュピタール バーゼル Immunoliposomes for the treatment of cancer

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH041122A (en) * 1990-04-16 1992-01-06 Olympus Optical Co Ltd Sustained release material and production thereof
US8333996B2 (en) * 1995-05-19 2012-12-18 Etex Corporation Calcium phosphate delivery vehicle and adjuvant
EP1874342B1 (en) * 2005-04-26 2018-06-06 Eisai R&D Management Co., Ltd. Compositions and methods for cancer immunotherapy
EP1984004A4 (en) * 2006-01-17 2010-03-03 Sloan Kettering Inst Cancer Therapy-enhancing glucan

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009024014A (en) * 2001-03-26 2009-02-05 Austin Research Inst Antibody against cancer
JP2009195739A (en) * 2004-10-14 2009-09-03 Biomimetic Therapeutics Inc Platelet-derived growth factor composition and method of use thereof
JP2007000637A (en) * 2005-06-22 2007-01-11 Heraeus Kulzer Gmbh Moldable implant material and the use
JP2009525048A (en) * 2006-02-01 2009-07-09 ザ ジョンズ ホプキンス ユニバーシティー Polypeptide-nucleic acid complex for immunological prophylaxis or immunotherapy against neoplastic disorders or infections
JP2007217307A (en) * 2006-02-14 2007-08-30 Meiji Univ Support for drug delivery and pharmaceutical using the same
JP2010540498A (en) * 2007-09-28 2010-12-24 ウニヴェルズィテーツシュピタール バーゼル Immunoliposomes for the treatment of cancer
JP2010126434A (en) * 2008-11-25 2010-06-10 Olympus Corp Cancer cell inhibitor and cancer cell-inhibiting sheet

Also Published As

Publication number Publication date
JP5990752B2 (en) 2016-09-14
JPWO2012105219A1 (en) 2014-07-03

Similar Documents

Publication Publication Date Title
Kanagavelu et al. In vivo effects of lattice radiation therapy on local and distant lung cancer: potential role of immunomodulation
CN108136025B (en) A novel method of treating cancer using immunomodulation
JP2022535610A (en) Compositions and methods for cancer immunotherapy
Chang et al. Combined GM‐CSF and IL‐12 gene therapy synergistically suppresses the growth of orthotopic liver tumors
RU2689160C2 (en) Anti-cancer targeted immunotherapy with application of il-12
CN106457001B (en) Methods for enhancing the efficacy of an immune response against a tumor
CA2652870C (en) Treatment of cancer with anti-il-1a antibodies
JP2023036900A (en) Antibody-drug conjugate preparation and lyophilization for the same
CN114159548A (en) Use of recombinant LAG-3 or derivatives thereof for inducing a monocyte immune response
JP2013529187A (en) Compositions comprising polypeptides such as polyglutamic acid nanoparticles and CD40 agonist
WO2021182573A1 (en) Medicament for treatment and/or prevention of cancer
WO2016021720A1 (en) Therapeutic agent for cancer which comprises combination of il-18 and molecule-targeting antibody
US20230263743A1 (en) Nanoparticle complex with defined sizes
JP6038486B2 (en) Drugs and nutritional supplements containing Taiwan Green Propolis Extract
WO2016119308A1 (en) Antitumour preparation and preparation method thereof
JP2010220479A (en) Method for culturing nk cell and use of the same
KR102606179B1 (en) Combinations containing ABX196 for the treatment of cancer
JP6082901B2 (en) Vaccine / adjuvant
JP5990752B2 (en) Antibody therapy effect enhancer
CN111166878B (en) Preparation method and application of combination of antibody targeting tumor antigen and iNKT cell
JP5966129B2 (en) Immunostimulator
WO2021182571A1 (en) Medicament for treatment and/or prevention of cancer
CN111225672B (en) Mevalonate pathway inhibitor and pharmaceutical composition thereof
JP5828470B2 (en) Immune inducer
CN110639013A (en) HPV nano vaccine composition with IL-33 as adjuvant and preparation method thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12742037

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2012555739

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 12742037

Country of ref document: EP

Kind code of ref document: A1