WO2022244854A1 - Anti-cd206 antibody and use thereof - Google Patents
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- WO2022244854A1 WO2022244854A1 PCT/JP2022/020889 JP2022020889W WO2022244854A1 WO 2022244854 A1 WO2022244854 A1 WO 2022244854A1 JP 2022020889 W JP2022020889 W JP 2022020889W WO 2022244854 A1 WO2022244854 A1 WO 2022244854A1
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
Definitions
- the present invention relates to an anti-CD206 antibody and its use in the prevention or treatment of impaired glucose tolerance, insulin resistance, diabetes, and the like.
- Diabetes mellitus is a group of diseases characterized by chronic hyperglycemia due to insufficient insulin action and various characteristic metabolic abnormalities. It causes serious debilitating complications. It is thought that the disease develops due to the combination of genetic predisposition and lifestyle factors that increase insulin resistance, such as high-fat diet, lack of exercise, and obesity.
- Resident macrophages in adipose tissue play an important role in maintaining tissue homeostasis, for example, regulating insulin sensitivity by secreting pro- or anti-inflammatory cytokines.
- the present inventors previously demonstrated that M2 macrophages in adipose tissue regulate whole-body glucose homeostasis by suppressing preadipocyte proliferation via the CD206/TGF ⁇ signaling pathway.
- CD206-positive cells in adipose tissue are mainly M2 macrophages, and removal of CD206-positive M2 macrophages increased the number of small adipocytes and improved insulin sensitivity throughout the body (Non-Patent Document 1).
- Non-Patent Document 2 when we created and analyzed genetically modified mice that can remove only M2 macrophages in the body at any time, we found that normal mice exhibit beige-coloring of subcutaneous fat in cold weather, but M2 macrophages should be removed. It has been clarified that this beige formation is more intensely observed, resulting in increased basal metabolism, decreased blood sugar level, and promoted insulin sensitivity (Non-Patent Document 2).
- removing M2 macrophages in adipose tissue may improve basal metabolism, improve insulin sensitivity, and prevent or treat impaired glucose tolerance. Therefore, there is a need for means to eliminate M2 macrophages in adipose tissue.
- an object of the present invention is to provide means for removing M2 macrophages in adipose tissue. More specifically, an antibody capable of binding to CD206 and removing CD206-positive M2 macrophages in adipose tissue, a CD206-positive M2 macrophage cell-killing agent comprising the antibody as an active ingredient, and a metabolism comprising the antibody as an active ingredient.
- An object of the present invention is to provide a pharmaceutical composition for preventing or treating disorders.
- the present inventors conducted intensive studies with the aim of developing an antibody that binds to CD206 and can eliminate CD206-positive M2 macrophages in adipose tissue. As a result, we succeeded in developing an antibody having such functions. They also found that the antibodies of the present invention bind to CD206, can eliminate CD206-positive M2 macrophages in adipose tissue, and have preventive and therapeutic effects on metabolic disorders. Based on such knowledge, the present invention was completed.
- the gist of the present invention is as follows.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is an antibody selected from: (a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30; an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31; (b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO
- a CD206-positive M2 macrophage cell-killing agent comprising, as an active ingredient, the antibody that specifically binds to CD206 or a functional fragment thereof according to [1] or [2].
- the CD206-positive M2 macrophage cell-killing agent according to [4] which has CD206-positive M2 macrophage cytotoxicity due to CDC activity.
- a pharmaceutical composition for preventing or treating metabolic disorders comprising the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof as an active ingredient.
- the pharmaceutical composition of [7] wherein the metabolic disorder is one or more selected from impaired glucose tolerance, insulin resistance and diabetes.
- a method of producing an antibody that specifically binds to CD206 comprising the step of culturing the host cell of [10] so that the antibody is produced.
- a method for preventing or treating a metabolic disorder comprising the step of administering the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof to a subject in need of prevention or treatment. .
- [14] Use of an antibody that specifically binds to CD206 or a functional fragment thereof for producing a pharmaceutical composition for preventing or treating metabolic disorders.
- a method for killing CD206-positive M2 macrophage cells comprising the step of administering to a subject the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof.
- the present invention provides an antibody that binds to CD206 and can eliminate CD206-positive M2 macrophages in adipose tissue.
- the present invention also provides a CD206-positive M2 macrophage cell-killing agent containing the antibody as an active ingredient, and a pharmaceutical composition for preventing or treating metabolic disorders containing the antibody as an active ingredient.
- FIG. 1 shows the results of flow cytometry detection of CD206-positive M2 macrophages by the anti-CD206 antibody of the present invention.
- FIG. 2 shows the evaluation results of the CDC activity of the anti-CD206 antibody of the present invention on CD206-positive cells.
- FIG. 3 shows the evaluation results of the ADCC activity of the anti-CD206 antibody of the present invention against CD206-positive cells.
- FIG. 4 shows the evaluation results (qPCR) of the cell-killing effect of the anti-CD206 antibody of the present invention on CD206-positive M2 macrophages in living tissue.
- FIG. 5 shows the evaluation results (FACS) of the cell-killing activity of the anti-CD206 antibody of the present invention on CD206-positive M2 macrophages in living tissues.
- FIG. 1 shows the results of flow cytometry detection of CD206-positive M2 macrophages by the anti-CD206 antibody of the present invention.
- FIG. 2 shows the evaluation results of the CDC activity of the anti-CD206 antibody of the present invention on
- FIG. 6 shows the evaluation results of the metabolic disorder-ameliorating action of the anti-CD206 antibody of the present invention.
- FIG. 7 is a diagram showing evaluation results of the effect of the anti-CD206 antibody of the present invention on improving metabolic disorders and the duration of the effect (results with different concentrations of the test antibody).
- FIG. 8 is a diagram showing evaluation results of the effect of the anti-CD206 antibody of the present invention on improving metabolic disorders and the duration of the effect (results of comparison between a test antibody and a target antibody).
- FIG. 9 shows the evaluation results of the mechanism of action of the anti-CD206 antibody of the present invention for improving metabolic disorders (improvement rate of blood glucose level before antibody administration and one week after antibody administration in each mouse).
- FIG. 10 shows the initial safety evaluation results (body weight, food intake) of the anti-CD206 antibody of the present invention.
- FIG. 11 shows the initial safety evaluation results (tissue weight) of the anti-CD206 antibody of the present invention.
- One aspect of the present invention is an antibody or a functional fragment thereof that specifically binds to CD206, wherein the antibody is an antibody selected from (a) to (k) above (hereinafter referred to as "the present invention (sometimes referred to as "anti-CD206 antibody”) or a functional fragment thereof.
- CD206 also known as the mannose receptor, is a C-type lectin receptor with a single transmembrane structure and a surface antigen specific to M2 macrophages.
- the present inventors have investigated means for removing M2 macrophages in adipose tissue, and have found that they bind to CD206 and have cell-killing activity against CD206-positive M2 macrophages in adipose tissue.
- An anti-CD206 antibody of the present invention capable of eliminating CD206-positive M2 macrophages was developed.
- the anti-CD206 antibody of the present invention having such action can be used as a pharmaceutical composition for the prevention or treatment of metabolic disorders. The present invention is thus completed.
- the anti-CD206 antibody of the present invention can be obtained, for example, by administering the antigen to an animal that produces an antibody against a specific antigen, immunizing the animal, collecting the antibody-producing cells, and then screening by the ISAAC (ImmunoSpot Array Assay on a Chip) method. be able to.
- the ISAAC method is a method for rapid and exhaustive screening of antigen-specific antibody-secreting cells (ASC) using a microwell array chip. In this method, a microwell array chip having a large number of wells large enough to accommodate one antibody-producing cell (about 10 to 15 ⁇ m in diameter), wherein the chip surface around the wells is coated with an anti-immunoglobulin antibody (or an anti-immunoglobulin antibody).
- microwell array chip coated with the antigen for which you are trying to obtain Splenic lymphocytes containing antibody-producing cells are collected from animals that produce antibodies against specific antigens, and the cells are seeded one by one in the wells of a microwell array chip.
- the chip is immersed in a culture medium and cultured under conditions in which the secreted antibodies diffuse from the wells to the coating layer, thereby producing antibodies. Binds globulin antibodies (or antigens).
- the antigen (or antibody against the antigen) labeled with a fluorescent substance is added to the microwell array chip.
- the labeled antigen (or antibody against the antigen) binds to the anti-immunoglobulin antibody (or antigen) around the well, and the signal emitted from the labeling substance stores cells that produce and secrete the antigen-specific antibody in which well. You can see if it is done.
- the cells are harvested using a microcapillary under a fluorescence microscope. DNA is extracted from cells, antibody cDNA is amplified by RT-PCR or the like, and antibody genes are cloned. A desired antibody can be produced as a recombinant antibody using the gene.
- lymphocytes are collected from the spleen of a CD206-deficient mouse to which the CD206 antigen was administered
- the cDNA of the mouse antibody against CD206 can be obtained by the above-mentioned ISAAC
- the mouse anti-human CD206 monoclonal antibody is a recombinant antibody.
- the ISAAC method is disclosed in JP-A-2009-34047, JP-A-2014-73100, JP-A-2014-162772, Aishun Jin et al., Nature Medicine, Vol.15, No. 9, September 2009, pp. 1088-1093 and Aishun Jin et al., Nature Protocols, Vol.6, No.5, 2011, pp.668-676.
- mice anti-human anti-CD206 monoclonal antibodies were obtained by the ISAAC method. No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83
- a monoclonal antibody having the above specific sequence can also be obtained using techniques commonly used in the field, such as chemical synthesis and genetic recombination, based on sequence information.
- the antibodies of the present invention include genetically modified antibodies artificially modified for the purpose of reducing heteroantigenicity against humans, such as chimeric antibodies, humanized antibodies, and humanized antibodies. Also included are antibodies, human antibodies, and the like. These antibodies can be produced using known methods. As used herein, the terms No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73 and 83 antibodies are used, except where specifically construed as such, are inconsistent. It includes chimeric antibodies and humanized antibodies made based on the sequences of the antibody.
- the present invention also includes shuffled antibodies containing the H chain or L chain of any of the antibodies Nos. 10, 27, 35, 40, 44, 47, 54, 59, 72, 73 and 83.
- the present invention relates to an isolated DNA encoding the anti-CD206 antibody of the present invention or a functional fragment thereof (hereinafter also referred to as "the DNA of the present invention").
- the anti-CD206 antibody of the present invention is produced by inserting the antibody heavy chain-encoding DNA and the light chain-encoding DNA into an expression vector, transforming host cells with the vectors, and culturing the host cells.
- the above-described heavy chain-encoding DNA and light chain-encoding DNA may be inserted into the same expression vector, and the vector may be used to transform host cells.
- the DNA encoding the strands may be inserted into separate vectors and the two vectors used to transform a host cell.
- the DNAs encoding the heavy and light chain variable regions may be inserted into a vector into which the DNAs encoding the heavy and light chain constant regions of a specific isotype have been previously inserted.
- the vector may also contain DNA encoding a signal peptide that facilitates secretion of the antibody from the host cell.
- the DNA encoding the signal peptide and the DNA encoding the antibody are ligated in-frame.
- the antibody can be obtained as a mature protein because the signal peptide is removed when the antibody is produced.
- the DNA and amino acid sequences of the heavy chain variable regions of 11 types of anti-CD206 monoclonal antibodies, the DNA and amino acid sequences of the light chain variable regions, the CDRs (complementarity determining regions) of the heavy chain variable regions (HCDR1, HCDR2 and HCDR3) The amino acid sequences and amino acid sequences or SEQ ID NOs of the light chain variable region CDRs (complementarity determining regions) (LCDR1, LCDR2 and LCDR3) are shown below.
- Antibody No. 10 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.27 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:41.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.35 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:46.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.40 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.44 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56.
- heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52
- heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53
- heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54
- a light chain LCDR1 consisting of the amino
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.47 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:61.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.54 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.59 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.72 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.73 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:86.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- Antibody No.83 An antibody or functional fragment thereof that specifically binds to CD206, An antibody or functional fragment thereof, wherein said antibody is: (k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:87; heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88; heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89; a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90; An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91.
- An antibody or functional fragment thereof that specifically binds to CD206 An antibody or functional fragment thereof, wherein said antibody is: (K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
- the heavy chain variable regions consist of not only the heavy chain variable regions consisting of the amino acid sequences represented by the respective SEQ ID NOs, but also 1 or several, for example 1 to 10, of the amino acid sequences. 1, 1 to 5, 1 to 3, 1 to 2, or 1 amino acid is deleted, substituted, or added, and the antibody heavy chain variable region activity, that is, human CD206 A heavy chain variable region consisting of a protein with binding activity is also included.
- the light chain variable region is not limited to the light chain variable region consisting of the amino acid sequence represented by each SEQ ID NO.
- a light chain consisting of a protein consisting of an amino acid sequence in which 3, 1 to 2, or 1 amino acid is deleted, substituted, or added, and having the activity of the light chain variable region of an antibody, ie, the binding activity to human CD206. Also includes variable regions.
- substitutions, deletions, and additions may be introduced into CDRs, but are preferably introduced into regions other than CDRs.
- the amino acid substitutions are preferably conservative substitutions in order to maintain the properties of the present invention.
- a conservative substitution means replacing an amino acid residue with another chemically similar amino acid residue so as not to substantially alter the activity of the peptide. For example, replacement of a hydrophobic residue with another hydrophobic residue, replacement of a polar residue with another polar residue having the same charge, and the like.
- Examples of functionally similar amino acids that may undergo such substitutions include non-polar (hydrophobic) amino acids such as alanine, valine, isoleucine, leucine, proline, tryptophan, phenylalanine, methionine, and the like.
- Polar (neutral) amino acids include glycine, serine, threonine, tyrosine, glutamine, asparagine, cysteine and the like.
- Examples of positively charged (basic) amino acids include arginine, histidine, and lysine.
- aspartic acid, glutamic acid, etc. are mentioned as an (acidic) amino acid with a negative charge.
- amino acid sequence of each SEQ ID NO and BLAST Basic Local Alignment Search Tool at the National Center for Biological Information (USA) Basic Local Alignment Search Tool of the National Center for Biological Information
- amino acid sequence of each SEQ ID NO and BLAST Basic Local Alignment Search Tool at the National Center for Biological Information (USA) Basic Local Alignment Search Tool of the National Center for Biological Information
- BLAST Basic Local Alignment Search Tool at the National Center for Biological Information (USA) Basic Local Alignment Search Tool of the National Center for Biological Information
- a protein having an amino acid sequence in which one or several amino acids are deleted, substituted or added in such an amino acid sequence is substantially identical to the protein having the amino acid sequence of each SEQ ID NO.
- the DNA encoding the amino acid sequence represented by each of the above SEQ ID NOs the DNA sequence and BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information (Basic Local Alignment Search Tool at the National Center for Biological Information) search tool) etc. (e.g. default i.e. default parameters), e.g. A DNA consisting of an identical nucleotide sequence and encoding a protein having the activity of an antibody heavy chain variable region or light chain variable region, that is, a protein having binding activity to human CD206 is also an anti-CD206 antibody. Included in the DNA encoding the heavy chain variable region or light chain variable region.
- a DNA capable of hybridizing under stringent conditions to a DNA comprising a sequence complementary to the DNA encoding the amino acid sequence represented by each of the above SEQ ID NOs, which is an antibody heavy chain variable region or DNAs encoding proteins having light chain variable region activity, ie, human CD206-binding activity, are also included in DNAs encoding heavy or light chain variable regions of anti-CD206 antibodies.
- SEQ ID NOs which is an antibody heavy chain variable region or DNAs encoding proteins having light chain variable region activity, ie, human CD206-binding activity
- a human anti-human CD206 monoclonal antibody that specifically binds to human CD206 comprising the heavy chain variable region and the light chain variable region includes the above heavy chain variable region and heavy chain constant region and the above light chain variable region and light chain constant region.
- the heavy chain constant region is composed of three domains, C H 1, C H 2 and C H 3.
- the heavy chain constant region may be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, but is most preferably an IgG1 or IgG4 constant region.
- the light chain constant region is composed of one domain, CL .
- Light chain constant regions are either kappa or lambda constant regions.
- the antibody-encoding DNA is formed by linking the heavy chain variable region-encoding DNA to the heavy chain constant region-encoding DNA, and further linking the light chain variable region-encoding DNA to the light chain constant region-encoding DNA.
- a heavy chain-encoding DNA and a light chain-encoding DNA are obtained.
- the antibodies of the present invention also include functional fragments of antibodies or modifications thereof.
- a functional fragment of an antibody is a fragment of an antibody that can specifically bind to an antigen.
- Functional fragments include Fab, F(ab') 2 , Fv, Fab/c having one Fab and a complete Fc, single-chain Fv ( scFv) and the like.
- Polynucleotides include both DNA and RNA.
- ⁇ Cell killer> Another aspect of the present invention relates to a CD206-positive M2 macrophage cell-killing agent (hereinafter sometimes referred to as "the cell-killing agent of the present invention") comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient. .
- the cell-killing agent of the present invention comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient.
- the anti-CD206 antibody of the present invention binds to CD206, preferably to the extracellular domain of CD206 present on the cell membrane surface of M2 macrophages, and has a cell-killing effect on CD206-positive M2 macrophages.
- the cell-killing action may be one or more selected from CDC activity and ADCC activity.
- the anti-CD206 antibody of the present invention has a cell-killing effect on CD206-positive M2 macrophages, can remove CD206-positive M2 macrophages from biological tissues, etc., improves glucose metabolism, and improves insulin sensitivity. and can be used to prevent or treat metabolic disorders.
- the cell-killing agent of the present invention is used as a reagent, the preparation, use, etc.
- the reagent can be carried out according to conventional methods in the field of molecular biology.
- the optional ingredient, dosage, target metabolic disorder, preventive or therapeutic target, etc. are described in the section ⁇ Pharmaceutical composition> below. It can also be applied in cell killing agents.
- ⁇ Pharmaceutical composition> Another aspect of the present invention is a pharmaceutical composition for the prevention or treatment of metabolic disorders, comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient (hereinafter referred to as the "pharmaceutical composition of the present invention"). (sometimes).
- any of the 11 mouse anti-human CD206 monoclonal antibodies of the present invention can be used as a pharmaceutical composition for preventing or treating metabolic disorders.
- antibodies Nos. 10, 35, 54 and 83, which strongly bind to CD206, are preferred, but not limited thereto.
- Antibodies can be used singly or in combination of any two or more.
- the pharmaceutical composition of the present invention can be carried out based on a conventional method in the pharmaceutical field.
- the pharmaceutical composition of the present invention may be used in combination with active ingredients other than the anti-CD206 antibody of the present invention as long as the effects are not impaired.
- the pharmaceutical composition may contain, in addition to the anti-CD206 antibody of the present invention, carriers, diluents and excipients commonly used in the pharmaceutical field. For example, lactose, magnesium stearate and the like are used as carriers and excipients for tablets.
- Aqueous solutions for injection include physiological saline, PBS, isotonic solutions containing glucose and other adjuvants, and suitable solubilizing agents, such as alcohol, polyalcohols such as propylene glycol, and nonionic interfaces. It may be used in combination with an active agent or the like. Sesame oil, soybean oil and the like are used as the oily liquid, and benzyl benzoate, benzyl alcohol and the like may be used together as the solubilizer.
- the pharmaceutical composition of the present invention can be administered in various forms, such as tablets, capsules, granules, powders, syrups, oral administration, injections, drip infusions, Parenteral administration via suppositories and the like can be mentioned.
- the dosage varies depending on the symptoms, age, body weight, etc., but in oral administration, it is usually about 0.01 mg to 1000 mg per day for adults, and these are administered once or in several divided doses. can do. In parenteral administration, for example, about 0.01 mg to 1000 mg can be administered by subcutaneous injection, intramuscular injection or intravenous injection.
- Metabolic disorders that may be prevented or treated include, but are not limited to, impaired glucose tolerance, insulin resistance, diabetes, diabetes-related diseases, metabolic syndrome, obesity, dyslipidemia, etc., preferably glucose tolerance function. disorders, insulin resistance, and diabetes.
- the diabetes more specifically includes prediabetes, type 1 diabetes, type 2 diabetes, insulin-resistant diabetes, glucose-intolerant diabetes, and the like, preferably type 2 diabetes.
- the diabetes-related diseases more specifically include hypertension, cardiovascular disease, renal disease, neuropathy, retinopathy, and the like.
- the prevention or treatment subject (preferably mammals, especially humans) in the present invention is a subject diagnosed with a high possibility of developing a metabolic disorder or a subject that has developed a metabolic disorder, and the pharmaceutical composition of the present invention is used. , can be administered to these subjects.
- the extracellular region of the mouse CD206 antigen 64-4161 of SEQ ID NO: 118 was expressed in cultured cells (FreeStyle (registered trademark) 293-F cells), and the purified protein was used as an antigen.
- CD206-deficient mouse B6.129P2-Mrc1 ⁇ tm1Mnz>/J mice were purchased from Jackson Laboratory.
- Example 1 Preparation of anti-CD206 monoclonal antibody> A mixture of CD206 antigen and adjuvant was administered to CD206-deficient mice for immunization. Two weeks after the first antibody administration, another CD206 antigen was administered, and spleens were collected one week later. Splenocytes were isolated and used to obtain CD206-specific antibodies using the ISAAC method, and the rest were stored at -80°C. First screening for lymphocytes producing antibodies that bind to CD206 was performed from spleen cells by the ISAAC method, and 90 clones were obtained. Antibody cDNA was cloned and transfected into HEK293F cells to obtain antibody-producing cells, and 13 types of mouse anti-human CD206 monoclonal antibodies were produced.
- the anti-CD206 antibody prepared as described above and the anti-CD206 antibody of the comparative example were detected by Allophycocyanin Labeling Kit-NH2 (detected by APC) (manufactured by DOJINDO).
- cultured cells HEK293Tcell
- FITC reporter-detected by FITC
- the anti-CD206 antibody of the present invention has the ability to bind to CD206.
- CDC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows. That is, CD206-positive fractions were collected by FACSAria (registered trademark) (manufactured by Nippon Becton Dickinson Co., Ltd.) and subjected to CDC assay. [CDC assay] 1. Collect spleen from 1 mouse 2. Homogenize using RPMI1640 3. Set Cell Strainer 70 ⁇ m in 50 mL Falcon (registered trademark) tube to collect isolated spleen cells 4. 4°C, 5 minutes, 1,500 Centrifuge at rpm 5.
- ADCC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows. Specifically, CD206-positive fractions were collected by FACSAria and subjected to ADCC assay. [ADCC assay] 1. Collect spleen from one mouse 2. Homogenize using RPMI1640 3. Set Cell Strainer 70 ⁇ m in 50mL Falcon tube to collect isolated spleen cells 4. Centrifuge at 1,500rpm for 5 minutes at 4°C 5. 4. Discard the supernatant and resuspend in 1x Lysis Buffer 6. Incubate at RT for 10 minutes 7.
- Example 4 Evaluation of cell-killing effect of anti-CD206 antibody on CD206-positive M2 macrophages in biological tissue>
- spleen, visceral fat, and subcutaneous fat were collected from the mice as follows, RNA was extracted, and qPCR was performed for F4/80, CD11c, and CD206 genes.
- spleen, visceral adipose (epididymal white adipose tissue: eWAT), and inguinal subcutaneous adipose (inguinal white adipose tissue: iWAT) cells were collected from mice and subjected to FACS analysis as follows. As a result, from the qPCR results shown in FIG.
- CD206-positive M2 macrophages tended to decrease in the visceral fat of the anti-CD206 antibody-administered group (indicated by arrows). Further, from the results of FACS analysis shown in FIG. 5, a significant decrease in CD206-positive M2 macrophages was confirmed in the visceral fat and subcutaneous fat of the anti-CD206 antibody-administered group. Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has a cell-killing effect on CD206-positive M2 macrophages in adipose tissue.
- qPCR 1. Harvest spleen, visceral fat, and subcutaneous fat from mice 2. RNA extraction with qiagen RNA extraction kit after tissue homogenization 3. Measure RNA concentration 4. Using TaKaRa Prime Script® ET reagent kit Generate cDNA (reverse transcription reaction) 37°C, 15 minutes (reverse transcription reaction) 85°C, 5 seconds (Heat inactivates reverse transcriptase) 4°C, ⁇ 5.
- KRHAG Krebs-Hense-leit-HEPES (KRHAG) buffer
- Physiological saline 5% BSA in physiological saline, 1M KCl, 1M CaCl2, 1M KH2PO4 , 1M MgSO4, 200mM Hepes, 200mM glucose (collagenase) 18mg in 9mL KRHAG (Available in advance and added just before use at -30°C)
- Example 5 Evaluation of metabolic disorder improving effect of anti-CD206 antibody>
- the anti-CD206 antibody of the present invention was evaluated for its ameliorating effect on metabolic disorders as follows.
- high-fat diet-loaded mice C57BL/6J mice (manufactured by CLEA Japan, Inc.) were fed a high-fat diet (reserch diets ultra-high-fat diet containing 60 kcal% fat and lard) from 8 weeks of age to 16 weeks as an obese diabetic model. A mouse was produced.
- the results are shown in Figure 6.
- the results of the glucose tolerance test showed that the anti-CD206 antibody produced in the present invention was administered to mice to improve impaired glucose tolerance.
- the results of insulin tolerance test showed that it improved insulin resistance. Therefore, it is believed that the anti-CD206 antibodies of the present invention can be used as pharmaceutical compositions for preventing or treating metabolic disorders.
- Figure 7 shows the results with different concentrations of the test antibody
- Figure 8 shows the comparison results of the test antibody and the target antibody.
- a single subcutaneous administration of the anti-CD206 antibody (30 ⁇ g/BW) prepared according to the present invention was confirmed to improve glucose tolerance for one week.
- Example 7 Elucidation of mechanism of action of anti-CD206 antibody to improve metabolic disorders> (CD206 knockout mouse) CD206 knockout mice were purchased from The Jackson Laboratory. The littermate mice were WT mice. Using these, high-fat diet loading was performed in the same manner as in Example 5.
- Liver liver
- eWAT visceral fat
- iWAT inguinal subcutaneous fat
- the results of body weight and food intake are shown in FIG. 10, and the results of tissue weight are shown in FIG.
- Administration of the anti-CD206 antibody continued for 28 days, but no sudden death, weight loss, decreased food consumption, or abnormal behavior was observed. A reduction in subcutaneous fat was confirmed. No other abnormal tissue findings were observed.
- the anti-CD206 antibody or functional fragment thereof of the present invention can be used as a pharmaceutical composition for prevention or treatment of impaired glucose tolerance, insulin resistance, diabetes, and the like.
- SEQ ID NO: 1 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 10)
- SEQ ID NO: 2 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 10)
- SEQ ID NO: 3 Anti-CD206 antibody (No. 11) heavy chain variable region amino acid sequence
- SEQ ID NO: 4 light chain variable region amino acid sequence of anti-CD206 antibody (No. 11)
- SEQ ID NO: 5 heavy chain variable region amino acid sequence of anti-CD206 antibody (No. 27)
- SEQ ID NO: 6 Amino acid sequence of light chain variable region of anti-CD206 antibody (No.
- SEQ ID NO: 7 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 35)
- SEQ ID NO: 8 Anti-CD206 antibody (No. 35)
- SEQ ID NO:9 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No.40)
- SEQ ID NO:10 Amino acid sequence of light chain variable region of anti-CD206 antibody (No.40)
- SEQ ID NO: 11 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 44)
- SEQ ID NO: 12 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 44)
- SEQ ID NO: 13 Anti-CD206 antibody (No.
- SEQ ID NO: 14 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 47)
- SEQ ID NO: 15 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 54)
- SEQ ID NO: 16 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 54)
- SEQ ID NO: 17 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 59)
- SEQ ID NO: 18 Anti-CD206 antibody (No. 59)
- SEQ ID NO: 20 Amino acid sequence of light chain variable region of anti-CD206 antibody (No.
- SEQ ID NO: 21 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 72)
- SEQ ID NO: 22 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 72)
- SEQ ID NO: 23 Anti-CD206 antibody (No. 73)
- SEQ ID NO: 24 Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 73)
- SEQ ID NO: 25 Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 83)
- SEQ ID NO: 27 Amino acid sequence of HCDR1 of anti-CD206 antibody (No.
- SEQ ID NO: 28 Anti-CD2 Amino acid sequence of HCDR2 of 06 antibody (No.10)
- SEQ ID NO:29 Amino acid sequence of HCDR3 of anti-CD206 antibody (No.10)
- SEQ ID NO:30 Amino acid sequence of LCDR1 of anti-CD206 antibody (No.10)
- SEQ ID NO:31 LCDR3 amino acid sequence of anti-CD206 antibody (No.10)
- SEQ ID NO:33 HCDR2 amino acid sequence of anti-CD206 antibody (No.11)
- SEQ ID NO:34 amino acid sequence of HCDR3 of anti-CD206 antibody (No.11)
- SEQ ID NO:35 amino acid sequence of LCDR1 of anti-CD206 antibody (No.11)
- SEQ ID NO:36 amino acid sequence of LCDR3 of anti-CD206 antibody (No.11)
- SEQ ID NO: 37 HCDR1 amino acid sequence of anti
- SEQ ID NO: 48 HCDR2 of anti-CD206 antibody (No. 40) Amino acid sequences SEQ ID NO: 49: Amino acid sequence of HCDR3 of anti-CD206 antibody (No. 40) SEQ ID NO: 50: Amino acid sequence of LCDR1 of anti-CD206 antibody (No. 40) SEQ ID NO: 51: LCDR3 of anti-CD206 antibody (No.
- SEQ ID NO: 52 HCDR1 amino acid sequence of anti-CD206 antibody (No.44)
- SEQ ID NO: 53 HCDR2 amino acid sequence of anti-CD206 antibody (No.44)
- SEQ ID NO: 54 anti-CD206 antibody (No.44)
- SEQ ID NO: 55 LCDR1 amino acid sequence of anti-CD206 antibody (No.44)
- SEQ ID NO: 78 HCDR2 amino acid sequence of anti-CD206 antibody (No. 72)
- SEQ ID NO: 79 anti-CD206 antibody (No. 72)
- SEQ ID NO: 80 LCDR1 amino acid sequence of anti-CD206 antibody (No. 72)
- SEQ ID NO: 81 LCDR3 amino acid sequence of anti-CD206 antibody (No. 72)
- SEQ ID NO: 94 Heavy chain variable region of anti-CD206 antibody (No. 11)
- SEQ ID NO:95 Nucleotide sequence encoding the light chain variable region of anti-CD206 antibody (No.11)
- SEQ ID NO:96 Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.27)
- No. 97 Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 27)
- SEQ ID NO: 98 Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.
- SEQ ID NO: 99 Anti-CD206 antibody Nucleotide sequence encoding the light chain variable region of (No.35) SEQ ID NO: 100: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.40) SEQ ID NO: 101: Anti-CD206 antibody (No.40) Nucleotide sequence encoding light chain variable region SEQ ID NO: 102: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 103: Encoding light chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 104: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.
- SEQ ID NO: 105 Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 47)
- SEQ ID NO: 106 Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.54)
- SEQ ID NO: 107 Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 54)
- SEQ ID NO: 108 Anti-CD206 antibody (No. 59)
- SEQ ID NO: 109 anti-CD206 antibody (No 59)
- SEQ ID NO: 110 Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No. 64)
- SEQ ID NO: 111 Light chain of anti-CD206 antibody (No.
- Nucleotide sequence encoding variable region SEQ ID NO: 112: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.72)
- SEQ ID NO: 113 Nucleotide encoding light chain variable region of anti-CD206 antibody (No.72)
- Sequence SEQ ID NO: 114 Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.73)
- SEQ ID NO: 115 Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No.73)
- SEQ ID NO: 117 Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 83)
- SEQ ID NO: 118 Nucleotide sequence of mouse CD206 coding region
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Abstract
The present invention addresses the problem of providing: an antibody which can bind to CD206 and can remove a CD206-positive M2 macrophage in an adipose tissue; an agent for killing a CD206-positive M2 macrophage cell; and a pharmaceutical composition for preventing or treating a metabolic disorder. The present invention provides: an antibody which can bind to CD206 and can remove a CD206-positive M2 macrophage in an adipose tissue; an agent for killing a CD206-positive M2 macrophage cell, the agent containing the antibody as an active ingredient; and a pharmaceutical composition for preventing or treating a metabolic disorder, the pharmaceutical composition containing the antibody as an active ingredient.
Description
本発明は、抗CD206抗体および該抗体の耐糖機能障害、インスリン抵抗性および糖尿病等の予防または治療用途に関する。
The present invention relates to an anti-CD206 antibody and its use in the prevention or treatment of impaired glucose tolerance, insulin resistance, diabetes, and the like.
糖尿病はインスリン作用の不足による慢性高血糖を主徴とし、様々の特徴的な代謝異常を伴う疾患群であり、病態の進行に伴い、眼、腎臓、神経、および血管を含む種々の器官に重篤な衰弱性合併症を引き起こす。
遺伝的素因に、高脂肪食・運動不足・肥満といったインスリン抵抗性増大をきたす生活環境因子が加わって発症するものと考えられている。 Diabetes mellitus is a group of diseases characterized by chronic hyperglycemia due to insufficient insulin action and various characteristic metabolic abnormalities. It causes serious debilitating complications.
It is thought that the disease develops due to the combination of genetic predisposition and lifestyle factors that increase insulin resistance, such as high-fat diet, lack of exercise, and obesity.
遺伝的素因に、高脂肪食・運動不足・肥満といったインスリン抵抗性増大をきたす生活環境因子が加わって発症するものと考えられている。 Diabetes mellitus is a group of diseases characterized by chronic hyperglycemia due to insufficient insulin action and various characteristic metabolic abnormalities. It causes serious debilitating complications.
It is thought that the disease develops due to the combination of genetic predisposition and lifestyle factors that increase insulin resistance, such as high-fat diet, lack of exercise, and obesity.
脂肪組織の常在性マクロファージは、組織恒常性の維持に重要な役割を担っていて、例えば、炎症性あるいは抗炎症性のサイトカインを分泌することによってインスリン感受性を調節する。本発明者らは、先に、脂肪組織のM2マクロファージが、CD206/TGFβシグナル伝達経路を介して前駆脂肪細胞の増殖を抑制することで、全身のグルコース恒常性を調節することを明らかにした。脂肪組織のCD206陽性細胞は主にM2マクロファージであり、CD206陽性M2マクロファージを除去すると小さな脂肪細胞の数が増加し全身のインスリン感受性が改善された（非特許文献１）。また、生体内のM2マクロファージだけを任意のタイミングで除去することができる遺伝子改変マウスを作成し解析したところ、寒冷時には正常のマウスで皮下脂肪のベージュ化が起こるが、M2マクロファージを除去しておくと、このベージュ化がより強く見られ、その結果基礎代謝が高まって血糖値が低下し、インスリン感受性が促進されることを明らかにした（非特許文献２）。
Resident macrophages in adipose tissue play an important role in maintaining tissue homeostasis, for example, regulating insulin sensitivity by secreting pro- or anti-inflammatory cytokines. The present inventors previously demonstrated that M2 macrophages in adipose tissue regulate whole-body glucose homeostasis by suppressing preadipocyte proliferation via the CD206/TGFβ signaling pathway. CD206-positive cells in adipose tissue are mainly M2 macrophages, and removal of CD206-positive M2 macrophages increased the number of small adipocytes and improved insulin sensitivity throughout the body (Non-Patent Document 1). In addition, when we created and analyzed genetically modified mice that can remove only M2 macrophages in the body at any time, we found that normal mice exhibit beige-coloring of subcutaneous fat in cold weather, but M2 macrophages should be removed. It has been clarified that this beige formation is more intensely observed, resulting in increased basal metabolism, decreased blood sugar level, and promoted insulin sensitivity (Non-Patent Document 2).
したがって、脂肪組織におけるM2マクロファージを除去することで、基礎代謝を向上し、インスリン感受性を改善し、耐糖障害を予防または治療できる可能性がある。したがって、脂肪組織におけるM2マクロファージを除去する手段が求められている。
Therefore, removing M2 macrophages in adipose tissue may improve basal metabolism, improve insulin sensitivity, and prevent or treat impaired glucose tolerance. Therefore, there is a need for means to eliminate M2 macrophages in adipose tissue.
前記の状況を鑑み、本発明は、脂肪組織におけるM2マクロファージを除去する手段を提供することを課題とする。より具体的には、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージを除去可能な抗体、該抗体を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤、および該抗体を有効成分として含む、代謝障害の予防または治療のための医薬組成物を提供することを課題とする。
In view of the above situation, an object of the present invention is to provide means for removing M2 macrophages in adipose tissue. More specifically, an antibody capable of binding to CD206 and removing CD206-positive M2 macrophages in adipose tissue, a CD206-positive M2 macrophage cell-killing agent comprising the antibody as an active ingredient, and a metabolism comprising the antibody as an active ingredient An object of the present invention is to provide a pharmaceutical composition for preventing or treating disorders.
本発明者等は、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージを除去可能な抗体を開発することを目的として鋭意検討を行った。その結果、このような機能を有する抗体の開発に成功した。そして、本発明の抗体は、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージを除去可能であり、代謝障害の予防および治療作用を有することを見出した。このような知見に基づき、本発明を完成した。
The present inventors conducted intensive studies with the aim of developing an antibody that binds to CD206 and can eliminate CD206-positive M2 macrophages in adipose tissue. As a result, we succeeded in developing an antibody having such functions. They also found that the antibodies of the present invention bind to CD206, can eliminate CD206-positive M2 macrophages in adipose tissue, and have preventive and therapeutic effects on metabolic disorders. Based on such knowledge, the present invention was completed.
すなわち、本発明の要旨は、以下のとおりである。
［１］ CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下から選択される抗体である、抗体またはその機能的断片：
（ａ）配列番号２７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号２８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号２９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号３０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号３１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｂ）配列番号３７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号３８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号３９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＬＭＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｃ）配列番号４２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｄ）配列番号４７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＦＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｅ）配列番号５２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＡＴで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｆ）配列番号５７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｇ）配列番号６２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＫＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｈ）配列番号６７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号７０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号７１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｉ）配列番号７７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号７８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号７９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｊ）配列番号８２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＮで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；並びに
（ｋ）配列番号８７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号９０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号９１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 That is, the gist of the present invention is as follows.
[1] An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is an antibody selected from:
(a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31;
(b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 41;
(c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 46;
(d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51;
(e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56;
(f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 61;
(g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66;
(h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71;
(i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81;
(j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 86; and (k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 87 ,
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91.
［１］ CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下から選択される抗体である、抗体またはその機能的断片：
（ａ）配列番号２７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号２８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号２９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号３０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号３１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｂ）配列番号３７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号３８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号３９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＬＭＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｃ）配列番号４２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｄ）配列番号４７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＦＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｅ）配列番号５２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＡＴで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｆ）配列番号５７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｇ）配列番号６２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＫＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｈ）配列番号６７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号７０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号７１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｉ）配列番号７７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号７８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号７９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｊ）配列番号８２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＮで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；並びに
（ｋ）配列番号８７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号９０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号９１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 That is, the gist of the present invention is as follows.
[1] An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is an antibody selected from:
(a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31;
(b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 41;
(c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 46;
(d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51;
(e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56;
(f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 61;
(g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66;
(h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71;
(i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81;
(j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 86; and (k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 87 ,
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91.
［２］ 前記抗体が以下から選択される抗体である、［１］に記載のCD206に特異的に結合する抗体またはその機能的断片：
（Ａ）配列番号１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｂ）配列番号５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｃ）配列番号７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｄ）配列番号９で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１０で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｅ）配列番号１１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｆ）配列番号１３で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１４で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｇ）配列番号１５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｈ）配列番号１７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｉ）配列番号２１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｊ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；並びに
（Ｋ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。
［３］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片をコードする、単離されたDNA。 [2] The antibody or functional fragment thereof that specifically binds to CD206 according to [1], wherein the antibody is an antibody selected from:
(A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; (K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
[3] An isolated DNA encoding the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof.
（Ａ）配列番号１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｂ）配列番号５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｃ）配列番号７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｄ）配列番号９で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１０で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｅ）配列番号１１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｆ）配列番号１３で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１４で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｇ）配列番号１５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｈ）配列番号１７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｉ）配列番号２１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｊ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；並びに
（Ｋ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。
［３］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片をコードする、単離されたDNA。 [2] The antibody or functional fragment thereof that specifically binds to CD206 according to [1], wherein the antibody is an antibody selected from:
(A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; (K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
[3] An isolated DNA encoding the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof.
［４］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤。
［５］ CDC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、［４］に記載のCD206陽性M2マクロファージ細胞殺傷剤。
［６］ ADCC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、［４］に記載のCD206陽性M2マクロファージ細胞殺傷剤。
［７］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を有効成分として含む、代謝障害の予防または治療のための医薬組成物。
［８］ 前記代謝障害が、耐糖機能障害、インスリン抵抗性および糖尿病から選択される１種以上である、［７］に記載の医薬組成物。 [4] A CD206-positive M2 macrophage cell-killing agent comprising, as an active ingredient, the antibody that specifically binds to CD206 or a functional fragment thereof according to [1] or [2].
[5] The CD206-positive M2 macrophage cell-killing agent according to [4], which has CD206-positive M2 macrophage cytotoxicity due to CDC activity.
[6] The CD206-positive M2 macrophage cell-killing agent according to [4], which has CD206-positive M2 macrophage cytotoxicity due to ADCC activity.
[7] A pharmaceutical composition for preventing or treating metabolic disorders, comprising the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof as an active ingredient.
[8] The pharmaceutical composition of [7], wherein the metabolic disorder is one or more selected from impaired glucose tolerance, insulin resistance and diabetes.
［５］ CDC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、［４］に記載のCD206陽性M2マクロファージ細胞殺傷剤。
［６］ ADCC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、［４］に記載のCD206陽性M2マクロファージ細胞殺傷剤。
［７］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を有効成分として含む、代謝障害の予防または治療のための医薬組成物。
［８］ 前記代謝障害が、耐糖機能障害、インスリン抵抗性および糖尿病から選択される１種以上である、［７］に記載の医薬組成物。 [4] A CD206-positive M2 macrophage cell-killing agent comprising, as an active ingredient, the antibody that specifically binds to CD206 or a functional fragment thereof according to [1] or [2].
[5] The CD206-positive M2 macrophage cell-killing agent according to [4], which has CD206-positive M2 macrophage cytotoxicity due to CDC activity.
[6] The CD206-positive M2 macrophage cell-killing agent according to [4], which has CD206-positive M2 macrophage cytotoxicity due to ADCC activity.
[7] A pharmaceutical composition for preventing or treating metabolic disorders, comprising the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof as an active ingredient.
[8] The pharmaceutical composition of [7], wherein the metabolic disorder is one or more selected from impaired glucose tolerance, insulin resistance and diabetes.
［９］ ［３］に記載のDNAを含むベクター。
［１０］ ［３］に記載のDNAまたは［１０］に記載のベクターを含む宿主細胞。
［１１］ CD206に特異的に結合する抗体を作製する方法であって、該抗体が産生されるように［１０］に記載の宿主細胞を培養する工程を含む、方法。
［１２］ 予防または治療を必要とする対象に、［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を投与する工程を含む、代謝障害の予防または治療方法。
［１３］ CD206に特異的に結合する抗体またはその機能的断片の、代謝障害の予防または治療のための使用。
［１４］ CD206に特異的に結合する抗体またはその機能的断片の、代謝障害の予防または治療のための医薬組成物の製造のための使用。
［１５］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を対象に投与する工程を含む、CD206陽性M2マクロファージ細胞殺傷方法。
［１６］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片の、CD206陽性M2マクロファージ細胞殺傷のための使用。
［１７］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片の、CD206陽性M2マクロファージ細胞殺傷剤の製造のための使用。 [9] A vector containing the DNA of [3].
[10] A host cell containing the DNA of [3] or the vector of [10].
[11] A method of producing an antibody that specifically binds to CD206, comprising the step of culturing the host cell of [10] so that the antibody is produced.
[12] A method for preventing or treating a metabolic disorder, comprising the step of administering the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof to a subject in need of prevention or treatment. .
[13] Use of an antibody that specifically binds to CD206 or a functional fragment thereof for prevention or treatment of metabolic disorders.
[14] Use of an antibody that specifically binds to CD206 or a functional fragment thereof for producing a pharmaceutical composition for preventing or treating metabolic disorders.
[15] A method for killing CD206-positive M2 macrophage cells, comprising the step of administering to a subject the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof.
[16] Use of the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof for killing CD206-positive M2 macrophage cells.
[17] Use of the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof for producing a CD206-positive M2 macrophage cell-killing agent.
［１０］ ［３］に記載のDNAまたは［１０］に記載のベクターを含む宿主細胞。
［１１］ CD206に特異的に結合する抗体を作製する方法であって、該抗体が産生されるように［１０］に記載の宿主細胞を培養する工程を含む、方法。
［１２］ 予防または治療を必要とする対象に、［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を投与する工程を含む、代謝障害の予防または治療方法。
［１３］ CD206に特異的に結合する抗体またはその機能的断片の、代謝障害の予防または治療のための使用。
［１４］ CD206に特異的に結合する抗体またはその機能的断片の、代謝障害の予防または治療のための医薬組成物の製造のための使用。
［１５］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片を対象に投与する工程を含む、CD206陽性M2マクロファージ細胞殺傷方法。
［１６］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片の、CD206陽性M2マクロファージ細胞殺傷のための使用。
［１７］ ［１］または［２］に記載のCD206に特異的に結合する抗体またはその機能的断片の、CD206陽性M2マクロファージ細胞殺傷剤の製造のための使用。 [9] A vector containing the DNA of [3].
[10] A host cell containing the DNA of [3] or the vector of [10].
[11] A method of producing an antibody that specifically binds to CD206, comprising the step of culturing the host cell of [10] so that the antibody is produced.
[12] A method for preventing or treating a metabolic disorder, comprising the step of administering the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof to a subject in need of prevention or treatment. .
[13] Use of an antibody that specifically binds to CD206 or a functional fragment thereof for prevention or treatment of metabolic disorders.
[14] Use of an antibody that specifically binds to CD206 or a functional fragment thereof for producing a pharmaceutical composition for preventing or treating metabolic disorders.
[15] A method for killing CD206-positive M2 macrophage cells, comprising the step of administering to a subject the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof.
[16] Use of the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof for killing CD206-positive M2 macrophage cells.
[17] Use of the antibody that specifically binds to CD206 of [1] or [2] or a functional fragment thereof for producing a CD206-positive M2 macrophage cell-killing agent.
本発明により、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージを除去可能な抗体が提供される。また、本発明により、該抗体を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤、および該抗体を有効成分として含む、代謝障害の予防または治療のための医薬組成物を提供が提供される。
The present invention provides an antibody that binds to CD206 and can eliminate CD206-positive M2 macrophages in adipose tissue. The present invention also provides a CD206-positive M2 macrophage cell-killing agent containing the antibody as an active ingredient, and a pharmaceutical composition for preventing or treating metabolic disorders containing the antibody as an active ingredient.
以下、本発明の実施の形態について、説明する。ただし、本発明は以下の好ましい実施形態に限定されず、本発明の範囲内で自由に変更することができるものである。なお、本明細書において、数値範囲を「下限～上限」で表現するものに関しては、上限は「以下」であっても「未満」であってもよく、下限は「以上」であっても「超」であってもよい。
Embodiments of the present invention will be described below. However, the present invention is not limited to the following preferred embodiments, and can be freely modified within the scope of the present invention. In this specification, regarding the numerical range expressed as "lower limit to upper limit", the upper limit may be "less than" or "less than", and the lower limit may be "more than" or " It may be "super".
＜抗体またはその機能的断片＞
本発明の一態様は、CD206に特異的に結合する抗体またはその機能的断片であって、前記抗体が前記（ａ）～（ｋ）から選択される抗体である、抗体（以下、「本発明の抗CD206抗体」ということがある）またはその機能的断片に関する。 <Antibody or functional fragment thereof>
One aspect of the present invention is an antibody or a functional fragment thereof that specifically binds to CD206, wherein the antibody is an antibody selected from (a) to (k) above (hereinafter referred to as "the present invention (sometimes referred to as "anti-CD206 antibody") or a functional fragment thereof.
本発明の一態様は、CD206に特異的に結合する抗体またはその機能的断片であって、前記抗体が前記（ａ）～（ｋ）から選択される抗体である、抗体（以下、「本発明の抗CD206抗体」ということがある）またはその機能的断片に関する。 <Antibody or functional fragment thereof>
One aspect of the present invention is an antibody or a functional fragment thereof that specifically binds to CD206, wherein the antibody is an antibody selected from (a) to (k) above (hereinafter referred to as "the present invention (sometimes referred to as "anti-CD206 antibody") or a functional fragment thereof.
CD206は、マンノース受容体としても知られる、１回膜貫通構造を有するＣ型レクチン受容体であり、M２マクロファージ特異的な表面抗原である。
上述のように、本発明者らは、脂肪組織におけるM2マクロファージを除去する手段を検討していたところ、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージに対し細胞殺傷活性を有し、脂肪組織におけるCD206陽性M2マクロファージを除去可能な本発明の抗CD206抗体を開発した。また、このような作用を有する本発明の抗CD206抗体は、代謝障害の予防または治療のための医薬組成物と用いられることを確認した。本発明はこのようにして完成されたものである。 CD206, also known as the mannose receptor, is a C-type lectin receptor with a single transmembrane structure and a surface antigen specific to M2 macrophages.
As described above, the present inventors have investigated means for removing M2 macrophages in adipose tissue, and have found that they bind to CD206 and have cell-killing activity against CD206-positive M2 macrophages in adipose tissue. An anti-CD206 antibody of the present invention capable of eliminating CD206-positive M2 macrophages was developed. In addition, it was confirmed that the anti-CD206 antibody of the present invention having such action can be used as a pharmaceutical composition for the prevention or treatment of metabolic disorders. The present invention is thus completed.
上述のように、本発明者らは、脂肪組織におけるM2マクロファージを除去する手段を検討していたところ、CD206に結合し、脂肪組織におけるCD206陽性M2マクロファージに対し細胞殺傷活性を有し、脂肪組織におけるCD206陽性M2マクロファージを除去可能な本発明の抗CD206抗体を開発した。また、このような作用を有する本発明の抗CD206抗体は、代謝障害の予防または治療のための医薬組成物と用いられることを確認した。本発明はこのようにして完成されたものである。 CD206, also known as the mannose receptor, is a C-type lectin receptor with a single transmembrane structure and a surface antigen specific to M2 macrophages.
As described above, the present inventors have investigated means for removing M2 macrophages in adipose tissue, and have found that they bind to CD206 and have cell-killing activity against CD206-positive M2 macrophages in adipose tissue. An anti-CD206 antibody of the present invention capable of eliminating CD206-positive M2 macrophages was developed. In addition, it was confirmed that the anti-CD206 antibody of the present invention having such action can be used as a pharmaceutical composition for the prevention or treatment of metabolic disorders. The present invention is thus completed.
本発明の抗CD206抗体は、例えば、特定の抗原に対する抗体を産生する動物に抗原を投与、免疫し、抗体産生細胞を回収し、その後、ISAAC(ImmunoSpot Array Assay on a Chip)法によるスクリーニングにより得ることができる。
ISAAC法は、マイクロウェルアレイチップを用いて抗原特異的抗体分泌細胞（ASC）を迅速かつ網羅的にスクリーニングする方法である。該方法においては、抗体産生細胞が１個入る程度の大きさ（直径10～15μm程度）のウェルを多数有するマイクロウェルアレイチップであって、ウェルの周辺のチップ表面を抗免疫グロブリン抗体（または抗体を得ようとする抗原）で被覆したマイクロウェルアレイチップを用いる。特定の抗原に対する抗体を産生する動物から抗体産生細胞を含む脾臓リンパ球を採取し、マイクロウェルアレイチップのウェルに細胞を１個ずつ播種する。培養液中にチップを浸し、分泌された抗体がウェルから前記被覆層に拡散する条件で培養することにより、抗体を産生分泌する細胞から分泌された抗体が、格納されたウェルの周囲の抗免疫グロブリン抗体（または抗原）に結合する。次いで、蛍光物質で標識した前記抗原（または抗原に対する抗体）をマイクロウェルアレイチップに添加する。標識した前記抗原（または抗原に対する抗体）がウェルの周囲の抗免疫グロブリン抗体（または抗原）に結合し、標識物質から発せられるシグナルにより、どのウェル内に抗原特異的抗体を産生分泌する細胞が格納されているかがわかる。この細胞を蛍光顕微鏡下でマイクロキャピラリーを用いて回収する。細胞からDNAを抽出し、RT-PCR等により抗体のcDNAを増幅し、抗体遺伝子をクローニングする。該遺伝子を用いて、所望の抗体をリコンビナント抗体として作製することができる。 The anti-CD206 antibody of the present invention can be obtained, for example, by administering the antigen to an animal that produces an antibody against a specific antigen, immunizing the animal, collecting the antibody-producing cells, and then screening by the ISAAC (ImmunoSpot Array Assay on a Chip) method. be able to.
The ISAAC method is a method for rapid and exhaustive screening of antigen-specific antibody-secreting cells (ASC) using a microwell array chip. In this method, a microwell array chip having a large number of wells large enough to accommodate one antibody-producing cell (about 10 to 15 μm in diameter), wherein the chip surface around the wells is coated with an anti-immunoglobulin antibody (or an anti-immunoglobulin antibody). Use a microwell array chip coated with the antigen for which you are trying to obtain Splenic lymphocytes containing antibody-producing cells are collected from animals that produce antibodies against specific antigens, and the cells are seeded one by one in the wells of a microwell array chip. The chip is immersed in a culture medium and cultured under conditions in which the secreted antibodies diffuse from the wells to the coating layer, thereby producing antibodies. Binds globulin antibodies (or antigens). Next, the antigen (or antibody against the antigen) labeled with a fluorescent substance is added to the microwell array chip. The labeled antigen (or antibody against the antigen) binds to the anti-immunoglobulin antibody (or antigen) around the well, and the signal emitted from the labeling substance stores cells that produce and secrete the antigen-specific antibody in which well. You can see if it is done. The cells are harvested using a microcapillary under a fluorescence microscope. DNA is extracted from cells, antibody cDNA is amplified by RT-PCR or the like, and antibody genes are cloned. A desired antibody can be produced as a recombinant antibody using the gene.
ISAAC法は、マイクロウェルアレイチップを用いて抗原特異的抗体分泌細胞（ASC）を迅速かつ網羅的にスクリーニングする方法である。該方法においては、抗体産生細胞が１個入る程度の大きさ（直径10～15μm程度）のウェルを多数有するマイクロウェルアレイチップであって、ウェルの周辺のチップ表面を抗免疫グロブリン抗体（または抗体を得ようとする抗原）で被覆したマイクロウェルアレイチップを用いる。特定の抗原に対する抗体を産生する動物から抗体産生細胞を含む脾臓リンパ球を採取し、マイクロウェルアレイチップのウェルに細胞を１個ずつ播種する。培養液中にチップを浸し、分泌された抗体がウェルから前記被覆層に拡散する条件で培養することにより、抗体を産生分泌する細胞から分泌された抗体が、格納されたウェルの周囲の抗免疫グロブリン抗体（または抗原）に結合する。次いで、蛍光物質で標識した前記抗原（または抗原に対する抗体）をマイクロウェルアレイチップに添加する。標識した前記抗原（または抗原に対する抗体）がウェルの周囲の抗免疫グロブリン抗体（または抗原）に結合し、標識物質から発せられるシグナルにより、どのウェル内に抗原特異的抗体を産生分泌する細胞が格納されているかがわかる。この細胞を蛍光顕微鏡下でマイクロキャピラリーを用いて回収する。細胞からDNAを抽出し、RT-PCR等により抗体のcDNAを増幅し、抗体遺伝子をクローニングする。該遺伝子を用いて、所望の抗体をリコンビナント抗体として作製することができる。 The anti-CD206 antibody of the present invention can be obtained, for example, by administering the antigen to an animal that produces an antibody against a specific antigen, immunizing the animal, collecting the antibody-producing cells, and then screening by the ISAAC (ImmunoSpot Array Assay on a Chip) method. be able to.
The ISAAC method is a method for rapid and exhaustive screening of antigen-specific antibody-secreting cells (ASC) using a microwell array chip. In this method, a microwell array chip having a large number of wells large enough to accommodate one antibody-producing cell (about 10 to 15 μm in diameter), wherein the chip surface around the wells is coated with an anti-immunoglobulin antibody (or an anti-immunoglobulin antibody). Use a microwell array chip coated with the antigen for which you are trying to obtain Splenic lymphocytes containing antibody-producing cells are collected from animals that produce antibodies against specific antigens, and the cells are seeded one by one in the wells of a microwell array chip. The chip is immersed in a culture medium and cultured under conditions in which the secreted antibodies diffuse from the wells to the coating layer, thereby producing antibodies. Binds globulin antibodies (or antigens). Next, the antigen (or antibody against the antigen) labeled with a fluorescent substance is added to the microwell array chip. The labeled antigen (or antibody against the antigen) binds to the anti-immunoglobulin antibody (or antigen) around the well, and the signal emitted from the labeling substance stores cells that produce and secrete the antigen-specific antibody in which well. You can see if it is done. The cells are harvested using a microcapillary under a fluorescence microscope. DNA is extracted from cells, antibody cDNA is amplified by RT-PCR or the like, and antibody genes are cloned. A desired antibody can be produced as a recombinant antibody using the gene.
本発明においては、例えば、CD206抗原を投与したCD206欠損マウスの脾臓からリンパ球を採取し、前記のISAACにより、CD206に対するマウス抗体のcDNAを得ることができ、マウス抗ヒトCD206モノクローナル抗体をリコンビナント抗体として作製することができる。
In the present invention, for example, lymphocytes are collected from the spleen of a CD206-deficient mouse to which the CD206 antigen was administered, the cDNA of the mouse antibody against CD206 can be obtained by the above-mentioned ISAAC, and the mouse anti-human CD206 monoclonal antibody is a recombinant antibody. can be made as
ISAAC法は、特開2009-34047号公報、特開2014-73100号公報、特開2014-162772号公報、Aishun Jin et al., Nature Medicine, Vol.15, No. 9, September 2009, pp.1088-1093およびAishun Jin et al., Nature Protocols, Vol.6, No.5, 2011, pp.668-676の記載に従って行うことができる。
The ISAAC method is disclosed in JP-A-2009-34047, JP-A-2014-73100, JP-A-2014-162772, Aishun Jin et al., Nature Medicine, Vol.15, No. 9, September 2009, pp. 1088-1093 and Aishun Jin et al., Nature Protocols, Vol.6, No.5, 2011, pp.668-676.
本発明では、ISAAC法により、下記の11種類のマウス抗ヒト抗CD206モノクローナル抗体が得られた。
No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83 In the present invention, the following 11 types of mouse anti-human anti-CD206 monoclonal antibodies were obtained by the ISAAC method.
No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83
No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83 In the present invention, the following 11 types of mouse anti-human anti-CD206 monoclonal antibodies were obtained by the ISAAC method.
No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83
上記の特定の配列を有するモノクローナル抗体は、配列情報に基づいて化学合成・遺伝子組み換え等の当分野で通常用いられている技術を用いて取得することもできる。
A monoclonal antibody having the above specific sequence can also be obtained using techniques commonly used in the field, such as chemical synthesis and genetic recombination, based on sequence information.
本発明の抗体には、前記モノクローナル抗体に加え、ヒトに対する異種抗原性を低下させること等を目的として人為的に改変した遺伝子組換え型抗体、例えば、キメラ（Chimeric）抗体、ヒト化（Humanized）抗体、ヒト抗体等も含まれる。これらの抗体は、既知の方法を用いて製造することができる。本明細書において、No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73および83抗体との語は、特にそのように解することが不整合である場合を除き、当該抗体の配列に基づき作製されたキメラ抗体及びヒト化抗体を含む。
In addition to the above-mentioned monoclonal antibodies, the antibodies of the present invention include genetically modified antibodies artificially modified for the purpose of reducing heteroantigenicity against humans, such as chimeric antibodies, humanized antibodies, and humanized antibodies. Also included are antibodies, human antibodies, and the like. These antibodies can be produced using known methods. As used herein, the terms No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73 and 83 antibodies are used, except where specifically construed as such, are inconsistent. It includes chimeric antibodies and humanized antibodies made based on the sequences of the antibody.
本発明は、No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73および83の抗体のいずれかのH鎖またはL鎖を含んだシャッフル抗体も包含する。
The present invention also includes shuffled antibodies containing the H chain or L chain of any of the antibodies Nos. 10, 27, 35, 40, 44, 47, 54, 59, 72, 73 and 83.
本発明の別の一態様は、本発明の抗CD206抗体またはその機能的断片をコードする、単離されたDNA（以下、「本発明のDNA」ともいう）に関する。
本発明の抗CD206抗体は、抗体の重鎖をコードするDNAおよび軽鎖をコードするDNAを発現ベクターに挿入し、該ベクターを用いて宿主細胞を形質転換し、該宿主細胞を培養して産生させることができる。この際、上記の重鎖をコードするDNAおよび軽鎖をコードするDNAを同じ発現ベクターに挿入し、該ベクターを用いて宿主細胞を形質転換してもよいし、重鎖をコードするDNAと軽鎖をコードするDNAを別々のベクターに挿入し、２つのベクターを用いて宿主細胞を形質転換してもよい。この際、特定のアイソタイプの重鎖定常領域および軽鎖定常領域をコードするDNAを予め挿入したベクターに重鎖可変領域および軽鎖可変領域をコードするDNAを挿入してもよい。また、該ベクターは宿主細胞からの抗体の分泌を促進するシグナルペプチドをコードするDNAを含んでいてもよい。この場合、シグナルペプチドをコードするDNAと抗体をコードするDNAをインフレームで連結するようにする。抗体が産生される際にシグナルペプチドが除去されるので、抗体を成熟タンパク質として得ることができる。 Another aspect of the present invention relates to an isolated DNA encoding the anti-CD206 antibody of the present invention or a functional fragment thereof (hereinafter also referred to as "the DNA of the present invention").
The anti-CD206 antibody of the present invention is produced by inserting the antibody heavy chain-encoding DNA and the light chain-encoding DNA into an expression vector, transforming host cells with the vectors, and culturing the host cells. can be made In this case, the above-described heavy chain-encoding DNA and light chain-encoding DNA may be inserted into the same expression vector, and the vector may be used to transform host cells. The DNA encoding the strands may be inserted into separate vectors and the two vectors used to transform a host cell. In this case, the DNAs encoding the heavy and light chain variable regions may be inserted into a vector into which the DNAs encoding the heavy and light chain constant regions of a specific isotype have been previously inserted. The vector may also contain DNA encoding a signal peptide that facilitates secretion of the antibody from the host cell. In this case, the DNA encoding the signal peptide and the DNA encoding the antibody are ligated in-frame. The antibody can be obtained as a mature protein because the signal peptide is removed when the antibody is produced.
本発明の抗CD206抗体は、抗体の重鎖をコードするDNAおよび軽鎖をコードするDNAを発現ベクターに挿入し、該ベクターを用いて宿主細胞を形質転換し、該宿主細胞を培養して産生させることができる。この際、上記の重鎖をコードするDNAおよび軽鎖をコードするDNAを同じ発現ベクターに挿入し、該ベクターを用いて宿主細胞を形質転換してもよいし、重鎖をコードするDNAと軽鎖をコードするDNAを別々のベクターに挿入し、２つのベクターを用いて宿主細胞を形質転換してもよい。この際、特定のアイソタイプの重鎖定常領域および軽鎖定常領域をコードするDNAを予め挿入したベクターに重鎖可変領域および軽鎖可変領域をコードするDNAを挿入してもよい。また、該ベクターは宿主細胞からの抗体の分泌を促進するシグナルペプチドをコードするDNAを含んでいてもよい。この場合、シグナルペプチドをコードするDNAと抗体をコードするDNAをインフレームで連結するようにする。抗体が産生される際にシグナルペプチドが除去されるので、抗体を成熟タンパク質として得ることができる。 Another aspect of the present invention relates to an isolated DNA encoding the anti-CD206 antibody of the present invention or a functional fragment thereof (hereinafter also referred to as "the DNA of the present invention").
The anti-CD206 antibody of the present invention is produced by inserting the antibody heavy chain-encoding DNA and the light chain-encoding DNA into an expression vector, transforming host cells with the vectors, and culturing the host cells. can be made In this case, the above-described heavy chain-encoding DNA and light chain-encoding DNA may be inserted into the same expression vector, and the vector may be used to transform host cells. The DNA encoding the strands may be inserted into separate vectors and the two vectors used to transform a host cell. In this case, the DNAs encoding the heavy and light chain variable regions may be inserted into a vector into which the DNAs encoding the heavy and light chain constant regions of a specific isotype have been previously inserted. The vector may also contain DNA encoding a signal peptide that facilitates secretion of the antibody from the host cell. In this case, the DNA encoding the signal peptide and the DNA encoding the antibody are ligated in-frame. The antibody can be obtained as a mature protein because the signal peptide is removed when the antibody is produced.
11種類の抗CD206モノクローナル抗体の重鎖可変領域のDNA配列およびアミノ酸配列、軽鎖可変領域のDNA配列およびアミノ酸配列、重鎖可変領域のCDR(相補性決定領域）（HCDR1、HCDR2およびHCDR3）のアミノ酸配列、並びに軽鎖可変領域のCDR(相補性決定領域）（LCDR1、LCDR2およびLCDR3）のアミノ酸配列の配列または配列番号を、以下に示す。
The DNA and amino acid sequences of the heavy chain variable regions of 11 types of anti-CD206 monoclonal antibodies, the DNA and amino acid sequences of the light chain variable regions, the CDRs (complementarity determining regions) of the heavy chain variable regions (HCDR1, HCDR2 and HCDR3) The amino acid sequences and amino acid sequences or SEQ ID NOs of the light chain variable region CDRs (complementarity determining regions) (LCDR1, LCDR2 and LCDR3) are shown below.
（抗体No.10）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ａ）配列番号２７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号２８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号２９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号３０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号３１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No. 10)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ａ）配列番号２７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号２８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号２９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号３０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号３１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No. 10)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ａ）配列番号１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ａ）配列番号１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.27）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｂ）配列番号３７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号３８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号３９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＬＭＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.27)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:41.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｂ）配列番号３７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号３８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号３９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＬＭＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.27)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:41.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｂ）配列番号５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｂ）配列番号５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.35）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｃ）配列番号４２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.35)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:46.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｃ）配列番号４２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.35)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:46.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｃ）配列番号７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｃ）配列番号７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.40）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｄ）配列番号４７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＦＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.40)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｄ）配列番号４７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＦＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.40)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｄ）配列番号９で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１０で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｄ）配列番号９で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１０で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.44）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｅ）配列番号５２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＡＴで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.44)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｅ）配列番号５２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＡＴで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.44)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｅ）配列番号１１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｅ）配列番号１１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.47）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｆ）配列番号５７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.47)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:61.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｆ）配列番号５７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.47)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:61.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｆ）配列番号１３で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１４で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｆ）配列番号１３で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１４で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.54）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｇ）配列番号６２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＫＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.54)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｇ）配列番号６２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＫＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.54)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｇ）配列番号１５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｇ）配列番号１５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.59）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｈ）配列番号６７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号７０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号７１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.59)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｈ）配列番号６７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号７０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号７１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.59)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｈ）配列番号１７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｈ）配列番号１７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.72）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｉ）配列番号７７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号７８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号７９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.72)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｉ）配列番号７７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号７８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号７９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.72)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｉ）配列番号２１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｉ）配列番号２１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.73）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｊ）配列番号８２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＮで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.73)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:86.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｊ）配列番号８２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＮで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.73)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:86.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｊ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｊ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
（抗体No.83）
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｋ）配列番号８７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号９０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号９１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.83)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:87;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（ｋ）配列番号８７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号９０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号９１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 (Antibody No.83)
An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:87;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91.
CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｋ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
前記抗体が以下の抗体である、抗体またはその機能的断片：
（Ｋ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is:
(K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity.
上記の11種類の抗CD206モノクローナル抗体において、重鎖可変領域はそれぞれの配列番号で表わされるアミノ酸配列からなる重鎖可変領域のみならず、該アミノ酸配列において、１もしくは数個、例えば、１～１０個、１～５個、１～３個、１～２個、または１個のアミノ酸が欠失、置換、付加されたアミノ酸配列からなり、抗体の重鎖可変領域の活性、すなわちヒトCD206への結合活性を有するタンパク質からなる重鎖可変領域も含む。同様に、軽鎖可変領域はそれぞれの配列番号で表わされるアミノ酸配列からなる軽鎖可変領域のみならず、該アミノ酸配列において１もしくは数個、例えば、１～１０個、１～５個、１～３個、１～２個、または１個のアミノ酸が欠失、置換、付加されたアミノ酸配列からなり、抗体の軽鎖可変領域の活性、すなわちヒトCD206への結合活性を有するタンパク質からなる軽鎖可変領域も含む。
In the 11 types of anti-CD206 monoclonal antibodies described above, the heavy chain variable regions consist of not only the heavy chain variable regions consisting of the amino acid sequences represented by the respective SEQ ID NOs, but also 1 or several, for example 1 to 10, of the amino acid sequences. 1, 1 to 5, 1 to 3, 1 to 2, or 1 amino acid is deleted, substituted, or added, and the antibody heavy chain variable region activity, that is, human CD206 A heavy chain variable region consisting of a protein with binding activity is also included. Similarly, the light chain variable region is not limited to the light chain variable region consisting of the amino acid sequence represented by each SEQ ID NO. A light chain consisting of a protein consisting of an amino acid sequence in which 3, 1 to 2, or 1 amino acid is deleted, substituted, or added, and having the activity of the light chain variable region of an antibody, ie, the binding activity to human CD206. Also includes variable regions.
このような置換、欠失、付加はCDRに導入されてもよいが、CDR以外の領域に導入されることが好ましい。また、該アミノ酸置換は本発明の特性を維持するために保存的置換であることが好ましい。ここで、保存的置換とは、ペプチドの活性を実質的に改変しないように、アミノ酸残基を、別の化学的に類似したアミノ酸残基で置換えることを意味する。例えば、ある疎水性残基を別の疎水性残基によって置換する場合、ある極性残基を同じ電荷を有する別の極性残基によって置換する場合等が挙げられる。このような置換を行うことができる機能的に類似のアミノ酸の例として、非極性（疎水性）アミノ酸としては、アラニン、バリン、イソロイシン、ロイシン、プロリン、トリプトファン、フェニルアラニン、メチオニン等が挙げられる。極性（中性）アミノ酸としては、グリシン、セリン、スレオニン、チロシン、グルタミン、アスパラギン、システイン等が挙げられる。陽電荷をもつ（塩基性）アミノ酸としては、アルギニン、ヒスチジン、リジン等が挙げられる。また、負電荷をもつ（酸性）アミノ酸としては、アスパラギン酸、グルタミン酸等が挙げられる。
Such substitutions, deletions, and additions may be introduced into CDRs, but are preferably introduced into regions other than CDRs. Also, the amino acid substitutions are preferably conservative substitutions in order to maintain the properties of the present invention. Here, a conservative substitution means replacing an amino acid residue with another chemically similar amino acid residue so as not to substantially alter the activity of the peptide. For example, replacement of a hydrophobic residue with another hydrophobic residue, replacement of a polar residue with another polar residue having the same charge, and the like. Examples of functionally similar amino acids that may undergo such substitutions include non-polar (hydrophobic) amino acids such as alanine, valine, isoleucine, leucine, proline, tryptophan, phenylalanine, methionine, and the like. Polar (neutral) amino acids include glycine, serine, threonine, tyrosine, glutamine, asparagine, cysteine and the like. Examples of positively charged (basic) amino acids include arginine, histidine, and lysine. Moreover, aspartic acid, glutamic acid, etc. are mentioned as an (acidic) amino acid with a negative charge.
このようなアミノ酸配列において１もしくは数個のアミノ酸が欠失、置換もしくは付加されたアミノ酸配列として、それぞれの配列番号のアミノ酸配列と、BLAST（Basic Local Alignment Search Tool at the National Center for Biological Information（米国国立生物学情報センターの基本ローカルアラインメント検索ツール））等（例えば、デフォルトすなわち初期設定のパラメータ）を用いて計算したときに、例えば、85％以上、90％以上、95％以上、97％以上、98％以上または99％以上の配列同一性を有しているものが挙げられる。
As an amino acid sequence in which one or several amino acids are deleted, substituted or added in such an amino acid sequence, the amino acid sequence of each SEQ ID NO and BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information (USA) Basic Local Alignment Search Tool of the National Center for Biological Information)), etc. (e.g., default or default parameters), e.g., 85% or more, 90% or more, 95% or more, 97% or more, Those having 98% or more or 99% or more sequence identity are included.
このようなアミノ酸配列において１もしくは数個のアミノ酸が欠失、置換もしくは付加されたアミノ酸配列を有するタンパク質はそれぞれの配列番号のアミノ酸配列を有するタンパク質と実質的に同一である。
A protein having an amino acid sequence in which one or several amino acids are deleted, substituted or added in such an amino acid sequence is substantially identical to the protein having the amino acid sequence of each SEQ ID NO.
また、上記のそれぞれの配列番号で表されるアミノ酸配列をコードするDNAとしては、該DNA配列とBLAST（Basic Local Alignment Search Tool at the National Center for Biological Information（米国国立生物学情報センターの基本ローカルアラインメント検索ツール））等（例えば、デフォルトすなわち初期設定のパラメータ）を用いて計算したときに、例えば、85％以上、90％以上、95％以上、97％以上、98％以上または99％以上の配列同一性を有している塩基配列からなるDNAであって、抗体の重鎖可変領域または軽鎖可変領域の活性、すなわちヒトCD206への結合活性を有するタンパク質をコードするDNAも、抗CD206抗体の重鎖可変領域または軽鎖可変領域をコードするDNAに含まれる。
In addition, as the DNA encoding the amino acid sequence represented by each of the above SEQ ID NOs, the DNA sequence and BLAST (Basic Local Alignment Search Tool at the National Center for Biological Information (Basic Local Alignment Search Tool at the National Center for Biological Information) search tool)) etc. (e.g. default i.e. default parameters), e.g. A DNA consisting of an identical nucleotide sequence and encoding a protein having the activity of an antibody heavy chain variable region or light chain variable region, that is, a protein having binding activity to human CD206 is also an anti-CD206 antibody. Included in the DNA encoding the heavy chain variable region or light chain variable region.
また、上記のそれぞれの配列番号で表されるアミノ酸配列をコードするDNAと相補的な配列からなるDNAとストリンジェントな条件下でハイブリダイズすることができるDNAであって抗体の重鎖可変領域または軽鎖可変領域の活性、すなわちヒトCD206への結合活性を有するタンパク質をコードするDNAも、抗CD206抗体の重鎖可変領域または軽鎖可変領域をコードするDNAに含まれる。ここで、当業者ならば、ストリンジェントな条件を適宜決定することができる。
Alternatively, a DNA capable of hybridizing under stringent conditions to a DNA comprising a sequence complementary to the DNA encoding the amino acid sequence represented by each of the above SEQ ID NOs, which is an antibody heavy chain variable region or DNAs encoding proteins having light chain variable region activity, ie, human CD206-binding activity, are also included in DNAs encoding heavy or light chain variable regions of anti-CD206 antibodies. Here, a person skilled in the art can appropriately determine stringent conditions.
また、前記の重鎖可変領域および軽鎖可変領域を含むヒトCD206に特異的に結合するヒト抗ヒトCD206モノクローナル抗体は、上記の重鎖可変領域および重鎖定常領域並びに上記の軽鎖可変領域および軽鎖定常領域とから構成される。重鎖定常領域は、３個のドメインCH1、CH2およびCH3から構成されている。重鎖定常領域は、IgG1、IgG2、IgG3、IgG4、IgA、IgE、IgMまたはIgD定常領域であってもよいが、最も好適には、IgG1またはIgG4定常領域である。軽鎖定常領域は、１個のドメインCLで構成されている。軽鎖定常領域は、κまたはλ定常領域である。
In addition, a human anti-human CD206 monoclonal antibody that specifically binds to human CD206 comprising the heavy chain variable region and the light chain variable region includes the above heavy chain variable region and heavy chain constant region and the above light chain variable region and light chain constant region. The heavy chain constant region is composed of three domains, C H 1, C H 2 and C H 3. The heavy chain constant region may be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, but is most preferably an IgG1 or IgG4 constant region. The light chain constant region is composed of one domain, CL . Light chain constant regions are either kappa or lambda constant regions.
抗体をコードするDNAは、重鎖可変領域をコードするDNAと重鎖定常領域をコードするDNAを連結し、さらに軽鎖可変領域をコードするDNAと軽鎖定常領域をコードするDNAを連結することにより重鎖をコードするDNAおよび軽鎖をコードするDNAとして得られる。
The antibody-encoding DNA is formed by linking the heavy chain variable region-encoding DNA to the heavy chain constant region-encoding DNA, and further linking the light chain variable region-encoding DNA to the light chain constant region-encoding DNA. A heavy chain-encoding DNA and a light chain-encoding DNA are obtained.
本発明の抗体は、抗体の機能的断片またはその修飾物も包含する。例えば、抗体の機能的断片は、抗体の断片であって抗原に特異的に結合し得る断片である。機能的断片としては、Fab、F(ab')2、Fv、1個のFabと完全なFcを有するFab/c、H鎖もしくはL鎖のFvを適当なリンカーで連結させたシングルチェインFv（scFv）等が挙げられる。ポリヌクレオチドは、DNAもRNAも包含する。
The antibodies of the present invention also include functional fragments of antibodies or modifications thereof. For example, a functional fragment of an antibody is a fragment of an antibody that can specifically bind to an antigen. Functional fragments include Fab, F(ab') 2 , Fv, Fab/c having one Fab and a complete Fc, single-chain Fv ( scFv) and the like. Polynucleotides include both DNA and RNA.
＜細胞殺傷剤＞
本発明の別の一態様は、本発明の抗CD206抗体またはその機能的断片を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤（以下、「本発明の細胞殺傷剤」ということがある）に関する。 <Cell killer>
Another aspect of the present invention relates to a CD206-positive M2 macrophage cell-killing agent (hereinafter sometimes referred to as "the cell-killing agent of the present invention") comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient. .
本発明の別の一態様は、本発明の抗CD206抗体またはその機能的断片を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤（以下、「本発明の細胞殺傷剤」ということがある）に関する。 <Cell killer>
Another aspect of the present invention relates to a CD206-positive M2 macrophage cell-killing agent (hereinafter sometimes referred to as "the cell-killing agent of the present invention") comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient. .
本発明の抗CD206抗体は、CD206、好ましくは、M2マクロファージの細胞膜表面に存在するCD206の細胞外領域に結合し、CD206陽性M2マクロファージに対する細胞殺傷作用を有する。細胞殺傷作用は、具体的には、CDC活性およびADCC活性から選ばれる1種以上であってよい。このように、本発明の抗CD206抗体は、CD206陽性M2マクロファージに対する細胞殺傷作用を有し、生体組織等からCD206陽性M2マクロファージを除去することができ、糖代謝を向上、およびインスリン感受性を改善することができ、代謝障害の予防または治療に用いることができる。本発明の細胞殺傷剤を試薬として使用する場合、試薬の製造、使用等は、分子生物学分野における常法に基づいて、行うことができる。本発明の細胞殺傷剤を医薬として使用する場合、任意成分、投与量、対象となる代謝障害、予防または治療対象等については、下記＜医薬組成物＞の項で説明した内容を、本発明の細胞殺傷剤においても適用することができる。
The anti-CD206 antibody of the present invention binds to CD206, preferably to the extracellular domain of CD206 present on the cell membrane surface of M2 macrophages, and has a cell-killing effect on CD206-positive M2 macrophages. Specifically, the cell-killing action may be one or more selected from CDC activity and ADCC activity. Thus, the anti-CD206 antibody of the present invention has a cell-killing effect on CD206-positive M2 macrophages, can remove CD206-positive M2 macrophages from biological tissues, etc., improves glucose metabolism, and improves insulin sensitivity. and can be used to prevent or treat metabolic disorders. When the cell-killing agent of the present invention is used as a reagent, the preparation, use, etc. of the reagent can be carried out according to conventional methods in the field of molecular biology. When the cell-killing agent of the present invention is used as a medicament, the optional ingredient, dosage, target metabolic disorder, preventive or therapeutic target, etc. are described in the section <Pharmaceutical composition> below. It can also be applied in cell killing agents.
＜医薬組成物＞
本発明の別の一態様は、本発明の抗CD206抗体またはその機能的断片を有効成分として含む、代謝障害の予防または治療のための医薬組成物（以下、「本発明の医薬組成物」ということがある）に関する。 <Pharmaceutical composition>
Another aspect of the present invention is a pharmaceutical composition for the prevention or treatment of metabolic disorders, comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient (hereinafter referred to as the "pharmaceutical composition of the present invention"). (sometimes).
本発明の別の一態様は、本発明の抗CD206抗体またはその機能的断片を有効成分として含む、代謝障害の予防または治療のための医薬組成物（以下、「本発明の医薬組成物」ということがある）に関する。 <Pharmaceutical composition>
Another aspect of the present invention is a pharmaceutical composition for the prevention or treatment of metabolic disorders, comprising the anti-CD206 antibody of the present invention or a functional fragment thereof as an active ingredient (hereinafter referred to as the "pharmaceutical composition of the present invention"). (sometimes).
本発明の11種類のマウス抗ヒトCD206モノクローナル抗体のいずれの抗体も代謝障害を予防もしくは治療するための医薬組成物として用いることができる。限定されないが、この中でも、CD206に強く結合する抗体No.10, 35, 54および83が好ましい。抗体は、１種のみまたは任意の２種以上を組み合わせて用いられ得る。
Any of the 11 mouse anti-human CD206 monoclonal antibodies of the present invention can be used as a pharmaceutical composition for preventing or treating metabolic disorders. Among these, antibodies Nos. 10, 35, 54 and 83, which strongly bind to CD206, are preferred, but not limited thereto. Antibodies can be used singly or in combination of any two or more.
本発明の医薬組成物の製造は製剤分野における常法に基づいて、行うことができる。なお、本発明の医薬組成物は、その効果を妨げない限り、本発明の抗CD206抗体以外の有効成分を併用し得る。該医薬組成物は、本発明の抗CD206抗体に加えて、製剤分野において通常用いられる担体、希釈剤、賦形剤を含み得る。例えば、錠剤用の担体、賦形剤としては、乳糖、ステアリン酸マグネシウム等が使用される。注射用の水性液としては、生理食塩水、PBS、ブドウ糖やその他の補助薬を含む等張液等が使用され、適当な溶解補助剤、例えば、アルコール、プロピレングリコール等のポリアルコール、非イオン界面活性剤等と併用してもよい。油性液としては、ゴマ油、大豆油等が使用され、溶解補助剤としては安息香酸ベンジル、ベンジルアルコール等を併用してもよい。本発明の医薬組成物は、種々の形態で投与することができ、それらの投与形態としては、錠剤、カプセル剤、顆粒剤、散剤、シロップ剤等による経口投与、または、注射剤、点滴剤、坐薬等による非経口投与を挙げることができる。
Manufacture of the pharmaceutical composition of the present invention can be carried out based on a conventional method in the pharmaceutical field. The pharmaceutical composition of the present invention may be used in combination with active ingredients other than the anti-CD206 antibody of the present invention as long as the effects are not impaired. The pharmaceutical composition may contain, in addition to the anti-CD206 antibody of the present invention, carriers, diluents and excipients commonly used in the pharmaceutical field. For example, lactose, magnesium stearate and the like are used as carriers and excipients for tablets. Aqueous solutions for injection include physiological saline, PBS, isotonic solutions containing glucose and other adjuvants, and suitable solubilizing agents, such as alcohol, polyalcohols such as propylene glycol, and nonionic interfaces. It may be used in combination with an active agent or the like. Sesame oil, soybean oil and the like are used as the oily liquid, and benzyl benzoate, benzyl alcohol and the like may be used together as the solubilizer. The pharmaceutical composition of the present invention can be administered in various forms, such as tablets, capsules, granules, powders, syrups, oral administration, injections, drip infusions, Parenteral administration via suppositories and the like can be mentioned.
その投与量は、症状、年齢、体重等によって異なるが、通常、経口投与では、成人に対して、例えば、1日約0.01mg～1000mgであり、これらを1回、または数回に分けて投与することができる。また、非経口投与では、例えば、1回約0.01mg～1000mgを皮下注射、筋肉注射または静脈注射によって投与することができる。
The dosage varies depending on the symptoms, age, body weight, etc., but in oral administration, it is usually about 0.01 mg to 1000 mg per day for adults, and these are administered once or in several divided doses. can do. In parenteral administration, for example, about 0.01 mg to 1000 mg can be administered by subcutaneous injection, intramuscular injection or intravenous injection.
予防または治療し得る代謝障害としては、限定されないが、耐糖機能障害、インスリン抵抗性、糖尿病、糖尿病に関連する疾患、メタボリック症候群、肥満症、および脂質異常症等が挙げられ、好ましくは、耐糖機能障害、インスリン抵抗性、および糖尿病である。限定されないが、前記糖尿病としてより具体的には、糖尿病前症、１型糖尿病、２型糖尿病、インスリン抵抗性糖尿病、およびグルコース不耐性糖尿病等が挙げられ、好ましくは２型糖尿病である。限定されないが、前記糖尿病に関連する疾患としてより具体的には、高血圧症、循環器病、腎疾患、神経障害、および網膜症等が挙げられる。
Metabolic disorders that may be prevented or treated include, but are not limited to, impaired glucose tolerance, insulin resistance, diabetes, diabetes-related diseases, metabolic syndrome, obesity, dyslipidemia, etc., preferably glucose tolerance function. disorders, insulin resistance, and diabetes. Although not limited, the diabetes more specifically includes prediabetes, type 1 diabetes, type 2 diabetes, insulin-resistant diabetes, glucose-intolerant diabetes, and the like, preferably type 2 diabetes. Although not limited, the diabetes-related diseases more specifically include hypertension, cardiovascular disease, renal disease, neuropathy, retinopathy, and the like.
本発明における予防または治療対象（好ましくは哺乳動物、特にヒト）は、代謝障害を発症する可能性が高いと診断された対象または代謝障害を発症した対象が対象となり、本発明の医薬組成物を、これらの対象に投与することができる。
The prevention or treatment subject (preferably mammals, especially humans) in the present invention is a subject diagnosed with a high possibility of developing a metabolic disorder or a subject that has developed a metabolic disorder, and the pharmaceutical composition of the present invention is used. , can be administered to these subjects.
以下、実施例により、本発明をさらに具体的に説明するが、本発明はその要旨を超えない限り、これらの実施例に限定されるものではない。
The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples as long as it does not exceed the gist thereof.
（抗原）
培養細胞（FreeStyle （登録商標） 293-F cell）にマウスCD206抗原の細胞外領域（配列番号１１８の64～4161）を発現させ、精製したタンパク質を抗原として用いた。
（CD206欠損マウス）
B6.129P2-Mrc1<tm1Mnz>/Jマウスは Jackson Laboratoryより購入した。 (antigen)
The extracellular region of the mouse CD206 antigen (64-4161 of SEQ ID NO: 118) was expressed in cultured cells (FreeStyle (registered trademark) 293-F cells), and the purified protein was used as an antigen.
(CD206-deficient mouse)
B6.129P2-Mrc1<tm1Mnz>/J mice were purchased from Jackson Laboratory.
培養細胞（FreeStyle （登録商標） 293-F cell）にマウスCD206抗原の細胞外領域（配列番号１１８の64～4161）を発現させ、精製したタンパク質を抗原として用いた。
（CD206欠損マウス）
B6.129P2-Mrc1<tm1Mnz>/Jマウスは Jackson Laboratoryより購入した。 (antigen)
The extracellular region of the mouse CD206 antigen (64-4161 of SEQ ID NO: 118) was expressed in cultured cells (FreeStyle (registered trademark) 293-F cells), and the purified protein was used as an antigen.
(CD206-deficient mouse)
B6.129P2-Mrc1<tm1Mnz>/J mice were purchased from Jackson Laboratory.
＜実施例１：抗CD206モノクローナル抗体の作製＞
CD206欠損マウスにCD206抗原とアジュバントを混合して投与し免疫を行った。１回目の抗体投与の２週間後にもう一度CD206抗原を投与し、その１週間後に脾臓を回収した。脾臓細胞は単離した後、ISAAC法を用いたCD206特異的抗体の取得に使用し、残りは-80℃で保存した。
脾臓細胞からISAAC法にて、CD206に結合する抗体を産生するリンパ球のファーストスクリーニングを行い、90クローンを得た。抗体cDNAをクローニングし、HEK293F細胞に遺伝子導入して、抗体産生細胞を得て、マウス抗ヒトCD206モノクローナル抗体を13種類作成した。 <Example 1: Preparation of anti-CD206 monoclonal antibody>
A mixture of CD206 antigen and adjuvant was administered to CD206-deficient mice for immunization. Two weeks after the first antibody administration, another CD206 antigen was administered, and spleens were collected one week later. Splenocytes were isolated and used to obtain CD206-specific antibodies using the ISAAC method, and the rest were stored at -80°C.
First screening for lymphocytes producing antibodies that bind to CD206 was performed from spleen cells by the ISAAC method, and 90 clones were obtained. Antibody cDNA was cloned and transfected into HEK293F cells to obtain antibody-producing cells, and 13 types of mouse anti-human CD206 monoclonal antibodies were produced.
CD206欠損マウスにCD206抗原とアジュバントを混合して投与し免疫を行った。１回目の抗体投与の２週間後にもう一度CD206抗原を投与し、その１週間後に脾臓を回収した。脾臓細胞は単離した後、ISAAC法を用いたCD206特異的抗体の取得に使用し、残りは-80℃で保存した。
脾臓細胞からISAAC法にて、CD206に結合する抗体を産生するリンパ球のファーストスクリーニングを行い、90クローンを得た。抗体cDNAをクローニングし、HEK293F細胞に遺伝子導入して、抗体産生細胞を得て、マウス抗ヒトCD206モノクローナル抗体を13種類作成した。 <Example 1: Preparation of anti-CD206 monoclonal antibody>
A mixture of CD206 antigen and adjuvant was administered to CD206-deficient mice for immunization. Two weeks after the first antibody administration, another CD206 antigen was administered, and spleens were collected one week later. Splenocytes were isolated and used to obtain CD206-specific antibodies using the ISAAC method, and the rest were stored at -80°C.
First screening for lymphocytes producing antibodies that bind to CD206 was performed from spleen cells by the ISAAC method, and 90 clones were obtained. Antibody cDNA was cloned and transfected into HEK293F cells to obtain antibody-producing cells, and 13 types of mouse anti-human CD206 monoclonal antibodies were produced.
上記のように作製した抗CD206抗体、および比較例の抗CD206抗体（市販の抗CD206抗体RATANTI MOUSE CD206（BIO-RAD社製）をAllophycocyanin Labeling Kit - NH2（APCで検出）(DOJINDO社製)で標識した。一方、培養細胞（HEK293Tcell）に遺伝子導入をしてCD206とVenus(FITCで検出)（富山大学分子神経科学講座作製）を発現させ、抗体のCD206陽性M2マクロファージへの結合能をフローサイトメトリーで検出した。結果は図１に示す。比較例の抗CD206抗体に対して、CD206陽性M2マクロファージを明瞭に検出することができた11種類（No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83）を最終的に選択した。
The anti-CD206 antibody prepared as described above and the anti-CD206 antibody of the comparative example (commercially available anti-CD206 antibody RATANTI MOUSE CD206 (manufactured by BIO-RAD) were detected by Allophycocyanin Labeling Kit-NH2 (detected by APC) (manufactured by DOJINDO). On the other hand, cultured cells (HEK293Tcell) were transfected with genes to express CD206 and Venus (detected by FITC) (prepared by the Department of Molecular Neuroscience, University of Toyama), and the ability of the antibody to bind to CD206-positive M2 macrophages was analyzed by flow cytometry. The results are shown in Figure 1. 11 types (No, 10, 27, 35, 40, 44, 47, 54, 59, 72, 73, 83) were finally selected.
さらに、CD206抗原（富山大学分子神経科学講座作製）を固定化したプレートを用いて、ELISAを行ったところ、抗CD206対照抗体（富山大学免疫学教室作製）を用いた場合にも認められる結合が、本発明の抗CD206抗体で結合が認められた。
したがって、本発明の抗CD206抗体がCD206に対する結合能を有することが示された。 Furthermore, when ELISA was performed using a plate on which the CD206 antigen (prepared by the Department of Molecular Neuroscience, University of Toyama) was immobilized, binding was also observed when using an anti-CD206 control antibody (prepared by the Department of Immunology, University of Toyama). , binding was observed with the anti-CD206 antibody of the present invention.
Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has the ability to bind to CD206.
したがって、本発明の抗CD206抗体がCD206に対する結合能を有することが示された。 Furthermore, when ELISA was performed using a plate on which the CD206 antigen (prepared by the Department of Molecular Neuroscience, University of Toyama) was immobilized, binding was also observed when using an anti-CD206 control antibody (prepared by the Department of Immunology, University of Toyama). , binding was observed with the anti-CD206 antibody of the present invention.
Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has the ability to bind to CD206.
13種類の抗体の重鎖および軽鎖の可変領域の塩基配列、並びに各CDRの塩基配列を解析した。同塩基配列に基づいて得られた抗体の重鎖および軽鎖可変領域のアミノ酸配列、並びに各CDRのアミノ酸配列を、下記の表１および配列表に示す。
We analyzed the base sequences of the heavy and light chain variable regions of 13 different antibodies, as well as the base sequences of each CDR. The amino acid sequences of the antibody heavy and light chain variable regions obtained based on the same nucleotide sequences, and the amino acid sequences of each CDR are shown in Table 1 and Sequence Listing below.
＜実施例２：抗CD206抗体の補体依存性細胞傷害(CDC: complement-dependent cytotoxicity)活性の評価＞
以下のようにして、本発明の抗CD206抗体のCD206陽性細胞に対するCDC活性を評価した。
すなわち、FACSAria（登録商標）（日本ベクトン・ディッキンソン株式会社製）にてCD206陽性分画を回収しCDC assayを行った。
［CDC assay］
1. マウス1匹から脾臓を採取
2. RPMI1640を使用してホモジナイズ
3. 50mL Falcon（登録商標）チューブにCell Strainer 70μmをセットし単離した脾臓細胞を回収する
4. 4℃、5分間、1,500rpmで遠心分離
5. 上清を捨て、1x Lysis Bufferで再懸濁する
6. RTで10分間インキュベート
7. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
8. CD3e-PE（for T cell） 7.5μL、CD19-APCCY7（for B cell） 5μL、NK1.1-FITC（for NK cell） 5μLを加え、40分間on iceで染色
9. FACS buffer 200μLを加え、1,500rpmで3分間遠心（96 well plate にて）
10. 上清を捨てる(10.～11.を3回繰り返す)
11. FACSAriaにてT cell等の染色細胞と無染色のCD206陽性分画を回収
12. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
13. 回収した細胞を96 wellプレートにまく
14. 抗CD206抗体または抗CD206対照抗体（Hel2, Hel6, Hel 10（富山大学免疫学教室作製）, rCD206（BIO-RAD社製）, MR（Abcam社製））を加える
15. 補体（Low Tox-M Rabbit complement（Cedarlane Laboratories Ltd 社製））を入れる（10% Comp）
16. 37℃ 1hrインキュベート
17. 二次抗体にて染色CD206陽性細胞（APC）・死細胞（7AAD）
18. FACSにてCD206陽性細胞と、CD206陽性かつ7AAD陽性細胞の割合を確認する <Example 2: Evaluation of complement-dependent cytotoxicity (CDC) activity of anti-CD206 antibody>
The CDC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows.
That is, CD206-positive fractions were collected by FACSAria (registered trademark) (manufactured by Nippon Becton Dickinson Co., Ltd.) and subjected to CDC assay.
[CDC assay]
1. Collect spleen from 1mouse 2. Homogenize using RPMI1640 3. Set Cell Strainer 70 μm in 50 mL Falcon (registered trademark) tube to collect isolated spleen cells 4. 4°C, 5 minutes, 1,500 Centrifuge at rpm 5. Discard supernatant and resuspend in 1x Lysis Buffer 6. Incubate at RT for 10 min 7. Centrifuge at 1,500 rpm for 5 min at 4°C and discard supernatant 8. CD3e-PE Add 7.5 μL (for T cell), 5 μL CD19-APCCY7 (for B cell), 5 μL NK1.1-FITC (for NK cell) and stain on ice for 40 minutes 9. Add 200 μL FACS buffer and rotate at 1,500 rpm for 3 Centrifugation for 1 minute (in a 96 well plate)
10. Discard the supernatant (repeat 10.-11. 3 times)
11. Collect stained cells such as T cells and unstained CD206-positive fraction by FACSAria
12. Centrifuge at 1,500 rpm for 5 minutes at 4℃ and discard the supernatant.
13. Spread the harvested cells on a 96-well plate
14. Add anti-CD206 antibody or anti-CD206 control antibody (Hel2, Hel6, Hel 10 (Prepared by Toyama University Immunology Department), rCD206 (BIO-RAD), MR (Abcam))
15. Add complement (Low Tox-M Rabbit complement (Cedarlane Laboratories Ltd)) (10% Comp)
16. Incubate at 37°C for 1 hour
17. CD206-positive cells (APC) and dead cells (7AAD) stained with secondary antibody
18. Confirm the ratio of CD206-positive cells and CD206-positive and 7AAD-positive cells by FACS
以下のようにして、本発明の抗CD206抗体のCD206陽性細胞に対するCDC活性を評価した。
すなわち、FACSAria（登録商標）（日本ベクトン・ディッキンソン株式会社製）にてCD206陽性分画を回収しCDC assayを行った。
［CDC assay］
1. マウス1匹から脾臓を採取
2. RPMI1640を使用してホモジナイズ
3. 50mL Falcon（登録商標）チューブにCell Strainer 70μmをセットし単離した脾臓細胞を回収する
4. 4℃、5分間、1,500rpmで遠心分離
5. 上清を捨て、1x Lysis Bufferで再懸濁する
6. RTで10分間インキュベート
7. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
8. CD3e-PE（for T cell） 7.5μL、CD19-APCCY7（for B cell） 5μL、NK1.1-FITC（for NK cell） 5μLを加え、40分間on iceで染色
9. FACS buffer 200μLを加え、1,500rpmで3分間遠心（96 well plate にて）
10. 上清を捨てる(10.～11.を3回繰り返す)
11. FACSAriaにてT cell等の染色細胞と無染色のCD206陽性分画を回収
12. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
13. 回収した細胞を96 wellプレートにまく
14. 抗CD206抗体または抗CD206対照抗体（Hel2, Hel6, Hel 10（富山大学免疫学教室作製）, rCD206（BIO-RAD社製）, MR（Abcam社製））を加える
15. 補体（Low Tox-M Rabbit complement（Cedarlane Laboratories Ltd 社製））を入れる（10% Comp）
16. 37℃ 1hrインキュベート
17. 二次抗体にて染色CD206陽性細胞（APC）・死細胞（7AAD）
18. FACSにてCD206陽性細胞と、CD206陽性かつ7AAD陽性細胞の割合を確認する <Example 2: Evaluation of complement-dependent cytotoxicity (CDC) activity of anti-CD206 antibody>
The CDC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows.
That is, CD206-positive fractions were collected by FACSAria (registered trademark) (manufactured by Nippon Becton Dickinson Co., Ltd.) and subjected to CDC assay.
[CDC assay]
1. Collect spleen from 1
10. Discard the supernatant (repeat 10.-11. 3 times)
11. Collect stained cells such as T cells and unstained CD206-positive fraction by FACSAria
12. Centrifuge at 1,500 rpm for 5 minutes at 4℃ and discard the supernatant.
13. Spread the harvested cells on a 96-well plate
14. Add anti-CD206 antibody or anti-CD206 control antibody (Hel2, Hel6, Hel 10 (Prepared by Toyama University Immunology Department), rCD206 (BIO-RAD), MR (Abcam))
15. Add complement (Low Tox-M Rabbit complement (Cedarlane Laboratories Ltd)) (10% Comp)
16. Incubate at 37°C for 1 hour
17. CD206-positive cells (APC) and dead cells (7AAD) stained with secondary antibody
18. Confirm the ratio of CD206-positive cells and CD206-positive and 7AAD-positive cells by FACS
結果を図２に示す。本発明で作製した抗CD206抗体を加えたwellではCD206陽性の生細胞の割合が減少し、CD206陽性の死細胞の割合が増加した。したがって、本発明の抗CD206抗体がCD206陽性細胞に対するCDC活性を有することが示された。
The results are shown in Figure 2. In the wells to which the anti-CD206 antibody prepared according to the present invention was added, the percentage of CD206-positive viable cells decreased and the percentage of CD206-positive dead cells increased. Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has CDC activity against CD206-positive cells.
＜実施例３：抗CD206抗体の抗体依存性細胞傷害（ADCC: antibody-dependent cell-mediated cytotoxicity）活性の評価＞
以下のようにして、本発明の抗CD206抗体のCD206陽性細胞に対するADCC活性を評価した。
すなわち、FACSAriaにてCD206陽性分画を回収しADCC assayを行った。
［ADCC assay］
1. マウス1匹から脾臓を採取
2. RPMI1640を使用してホモジナイズ
3. 50mL FalconチューブにCell Strainer 70μmをセットし単離した脾臓細胞を回収する
4. 4℃、5分間、1,500rpmで遠心分離
5. 4.上清を捨て、1x Lysis Bufferで再懸濁する
6. RTで10分間インキュベート
7. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
8. CD3e-PE 7.5μL、CD19-APCCY7 5μL、NK1.1-FITC 5μLを加え、40分間on iceで染色
9. FACS buffer 200μLを加え、1500rpmで3分間遠心（96 well plate にて）
10. 上清を捨てる(10.～11.を3回繰り返す)
11. FACSAriaにてT cell等の染色細胞と無染色のCD206陽性分画を回収
12. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
13. 回収した細胞を96 wellプレートにまく
14. 抗CD206抗体または抗CD206対照抗体を加える
15. 37℃ 12hrインキュベート
16. 二次抗体にて染色CD206陽性細胞（APC）・死細胞（7AAD）
17. FACSにてCD206陽性細胞と、CD206陽性かつ7AAD陽性細胞の割合を確認する <Example 3: Evaluation of antibody-dependent cell-mediated cytotoxicity (ADCC) activity of anti-CD206 antibody>
The ADCC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows.
Specifically, CD206-positive fractions were collected by FACSAria and subjected to ADCC assay.
[ADCC assay]
1. Collect spleen from onemouse 2. Homogenize using RPMI1640 3. Set Cell Strainer 70μm in 50mL Falcon tube to collect isolated spleen cells 4. Centrifuge at 1,500rpm for 5 minutes at 4℃ 5. 4. Discard the supernatant and resuspend in 1x Lysis Buffer 6. Incubate at RT for 10 minutes 7. Centrifuge at 1,500 rpm for 5 minutes at 4°C and discard the supernatant 8. CD3e-PE 7.5 μL , CD19-APCCY7 5 μL, NK1.1-FITC 5 μL, stained on ice for 40 minutes 9. Add FACS buffer 200 μL, centrifuge at 1500 rpm for 3 minutes (96 well plate)
10. Discard the supernatant (repeat 10.-11. 3 times)
11. Collect stained cells such as T cells and unstained CD206-positive fraction by FACSAria
12. Centrifuge at 1,500 rpm for 5 minutes at 4℃ and discard the supernatant.
13. Spread the harvested cells on a 96-well plate
14. Add anti-CD206 antibody or anti-CD206 control antibody
15. Incubate at 37°C for 12 hours
16. CD206-positive cells (APC) and dead cells (7AAD) stained with secondary antibody
17. Confirm the ratio of CD206-positive cells and CD206-positive and 7AAD-positive cells by FACS
以下のようにして、本発明の抗CD206抗体のCD206陽性細胞に対するADCC活性を評価した。
すなわち、FACSAriaにてCD206陽性分画を回収しADCC assayを行った。
［ADCC assay］
1. マウス1匹から脾臓を採取
2. RPMI1640を使用してホモジナイズ
3. 50mL FalconチューブにCell Strainer 70μmをセットし単離した脾臓細胞を回収する
4. 4℃、5分間、1,500rpmで遠心分離
5. 4.上清を捨て、1x Lysis Bufferで再懸濁する
6. RTで10分間インキュベート
7. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
8. CD3e-PE 7.5μL、CD19-APCCY7 5μL、NK1.1-FITC 5μLを加え、40分間on iceで染色
9. FACS buffer 200μLを加え、1500rpmで3分間遠心（96 well plate にて）
10. 上清を捨てる(10.～11.を3回繰り返す)
11. FACSAriaにてT cell等の染色細胞と無染色のCD206陽性分画を回収
12. 4℃、5分間、1,500rpmで遠心分離し、上清を捨てる
13. 回収した細胞を96 wellプレートにまく
14. 抗CD206抗体または抗CD206対照抗体を加える
15. 37℃ 12hrインキュベート
16. 二次抗体にて染色CD206陽性細胞（APC）・死細胞（7AAD）
17. FACSにてCD206陽性細胞と、CD206陽性かつ7AAD陽性細胞の割合を確認する <Example 3: Evaluation of antibody-dependent cell-mediated cytotoxicity (ADCC) activity of anti-CD206 antibody>
The ADCC activity of the anti-CD206 antibodies of the present invention against CD206-positive cells was evaluated as follows.
Specifically, CD206-positive fractions were collected by FACSAria and subjected to ADCC assay.
[ADCC assay]
1. Collect spleen from one
10. Discard the supernatant (repeat 10.-11. 3 times)
11. Collect stained cells such as T cells and unstained CD206-positive fraction by FACSAria
12. Centrifuge at 1,500 rpm for 5 minutes at 4℃ and discard the supernatant.
13. Spread the harvested cells on a 96-well plate
14. Add anti-CD206 antibody or anti-CD206 control antibody
15. Incubate at 37°C for 12 hours
16. CD206-positive cells (APC) and dead cells (7AAD) stained with secondary antibody
17. Confirm the ratio of CD206-positive cells and CD206-positive and 7AAD-positive cells by FACS
結果を図３に示す。本発明で作製した抗CD206抗体を加えたwellではCD206陽性の生細胞の割合が減少し、CD206陽性の死細胞の割合が増加した。したがって、本発明の抗CD206抗体がCD206陽性細胞に対するADCC活性を有することが示された。
The results are shown in Figure 3. In the wells to which the anti-CD206 antibody prepared according to the present invention was added, the percentage of CD206-positive viable cells decreased and the percentage of CD206-positive dead cells increased. Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has ADCC activity against CD206-positive cells.
＜実施例４：抗CD206抗体の生体組織におけるCD206陽性M2マクロファージに対する細胞殺傷作用の評価＞
以下のようにして、本発明の抗CD206抗体の生体組織におけるCD206陽性M2マクロファージに対する細胞殺傷作用を評価した。
C57BL/6Jマウス（日本クレア株式会社製)に、PBSに希釈した被験抗体および対照としてPBSを、それぞれ1μg/マウス体重(g)の濃度で48時間毎に2回腹腔内に投与した（No.47 n=4, No. 83 n=5）。投与4日（96時間）後、以下のようにして脾臓・内臓脂肪・皮下脂肪をマウスから回収しRNAを抽出し、F4/80、CD11c、CD206遺伝子についてqPCRを行った。また、以下のようにして脾臓（spleen）・内臓脂肪（epididymal white adipose tissue: eWAT）・鼠径部皮下脂肪（inguinal white adipose tissue: iWAT）細胞をマウスから回収し、FACS解析を行った。
その結果、図４に示すqPCRによる結果より、抗CD206抗体投与群の内臓脂肪においてCD206陽性M2マクロファージの減少傾向が確認できた（矢印で示す）。また、図５に示すFACS解析による結果より、抗CD206抗体投与群の内臓脂肪・皮下脂肪においてCD206陽性M2マクロファージの有意な減少が確認できた。したがって、本発明の抗CD206抗体が脂肪組織におけるCD206陽性M2マクロファージに対する細胞殺傷作用を有することが示された。 <Example 4: Evaluation of cell-killing effect of anti-CD206 antibody on CD206-positive M2 macrophages in biological tissue>
The cell-killing effect of the anti-CD206 antibody of the present invention on CD206-positive M2 macrophages in living tissue was evaluated as follows.
To C57BL/6J mice (manufactured by CLEA Japan, Inc.), the test antibody diluted in PBS and PBS as a control were intraperitoneally administered twice every 48 hours at a concentration of 1 μg/mouse body weight (g) (No. 47 n=4, No. 83 n=5). Four days (96 hours) after administration, spleen, visceral fat, and subcutaneous fat were collected from the mice as follows, RNA was extracted, and qPCR was performed for F4/80, CD11c, and CD206 genes. In addition, spleen, visceral adipose (epididymal white adipose tissue: eWAT), and inguinal subcutaneous adipose (inguinal white adipose tissue: iWAT) cells were collected from mice and subjected to FACS analysis as follows.
As a result, from the qPCR results shown in FIG. 4, it was confirmed that CD206-positive M2 macrophages tended to decrease in the visceral fat of the anti-CD206 antibody-administered group (indicated by arrows). Further, from the results of FACS analysis shown in FIG. 5, a significant decrease in CD206-positive M2 macrophages was confirmed in the visceral fat and subcutaneous fat of the anti-CD206 antibody-administered group. Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has a cell-killing effect on CD206-positive M2 macrophages in adipose tissue.
以下のようにして、本発明の抗CD206抗体の生体組織におけるCD206陽性M2マクロファージに対する細胞殺傷作用を評価した。
C57BL/6Jマウス（日本クレア株式会社製)に、PBSに希釈した被験抗体および対照としてPBSを、それぞれ1μg/マウス体重(g)の濃度で48時間毎に2回腹腔内に投与した（No.47 n=4, No. 83 n=5）。投与4日（96時間）後、以下のようにして脾臓・内臓脂肪・皮下脂肪をマウスから回収しRNAを抽出し、F4/80、CD11c、CD206遺伝子についてqPCRを行った。また、以下のようにして脾臓（spleen）・内臓脂肪（epididymal white adipose tissue: eWAT）・鼠径部皮下脂肪（inguinal white adipose tissue: iWAT）細胞をマウスから回収し、FACS解析を行った。
その結果、図４に示すqPCRによる結果より、抗CD206抗体投与群の内臓脂肪においてCD206陽性M2マクロファージの減少傾向が確認できた（矢印で示す）。また、図５に示すFACS解析による結果より、抗CD206抗体投与群の内臓脂肪・皮下脂肪においてCD206陽性M2マクロファージの有意な減少が確認できた。したがって、本発明の抗CD206抗体が脂肪組織におけるCD206陽性M2マクロファージに対する細胞殺傷作用を有することが示された。 <Example 4: Evaluation of cell-killing effect of anti-CD206 antibody on CD206-positive M2 macrophages in biological tissue>
The cell-killing effect of the anti-CD206 antibody of the present invention on CD206-positive M2 macrophages in living tissue was evaluated as follows.
To C57BL/6J mice (manufactured by CLEA Japan, Inc.), the test antibody diluted in PBS and PBS as a control were intraperitoneally administered twice every 48 hours at a concentration of 1 μg/mouse body weight (g) (No. 47 n=4, No. 83 n=5). Four days (96 hours) after administration, spleen, visceral fat, and subcutaneous fat were collected from the mice as follows, RNA was extracted, and qPCR was performed for F4/80, CD11c, and CD206 genes. In addition, spleen, visceral adipose (epididymal white adipose tissue: eWAT), and inguinal subcutaneous adipose (inguinal white adipose tissue: iWAT) cells were collected from mice and subjected to FACS analysis as follows.
As a result, from the qPCR results shown in FIG. 4, it was confirmed that CD206-positive M2 macrophages tended to decrease in the visceral fat of the anti-CD206 antibody-administered group (indicated by arrows). Further, from the results of FACS analysis shown in FIG. 5, a significant decrease in CD206-positive M2 macrophages was confirmed in the visceral fat and subcutaneous fat of the anti-CD206 antibody-administered group. Therefore, it was demonstrated that the anti-CD206 antibody of the present invention has a cell-killing effect on CD206-positive M2 macrophages in adipose tissue.
［qPCR］
1. 脾臓・内臓脂肪・皮下脂肪をマウスから回収
2. 組織をホモジナイズした後、qiagen RNA抽出キットでRNA抽出
3. RNA濃度を測定
4. TaKaRa Prime Script（登録商標） ET reagent kitを使用して cDNAを作製
（逆転写反応）
37℃、 15分 （逆転写反応）
85℃、 5秒 （逆転写酵素を熱失活させる）
4℃、 ∞
5. Real Time PCR (SYBR)
(1) 氷とサンプル(cDNA)を準備する
(2) コントロールと調べるプライマーを-20℃の冷凍庫から出す
アルミホイルの上に置く
(3) マスターMixを -20℃の冷凍庫から出して氷上におく
(4) Micro Ampのtubeと蓋を用意しておく
(5) 薄黄色のtubeをプライマー数分準備し、蓋にプライマーの名前を書いておく
(6) 用意した薄黄色のtubeにマスターMixとプライマーのMixを作る
マスターMix 12.5μL（7.5） × サンプル数 ＝ μL
H2O , PCR Grade 9μL（5.4） × サンプル数 ＝ μL
プライマー FW 0.5μL （0.4） × サンプル数 = μL
プライマー RV 0.5μL （0.4） × サンプル数 = μL
ROX Reference Dye II 0.5μL（0.3） × サンプル数 = μL
※マスターMixを入れたtubeに、プライマーをピペッティングしながら加える。
よく混ぜた後、スピンダウンする。
(7) 作ったMixをMicro Ampのtubeに23μLずつ分注する。
(8) Mix 23μLずつ分注したら、サンプルを2μLずつ加えて、合計25μLにする。
(7) 23μL ＋ (8) 2μL ＝ 25μL
(9) Micro Ampに蓋をして、泡がないか、蓋がきちんとしまっているかを確認したのち、スピンダウンして液を側面から落とす。
(10) リアルタイム定量PCRシステム（MX3000P（アジレント・テクノロジー株式会社製））にセットする。
6. MX3000Pの結果を解析する [qPCR]
1. Harvest spleen, visceral fat, and subcutaneous fat frommice 2. RNA extraction with qiagen RNA extraction kit after tissue homogenization 3. Measure RNA concentration 4. Using TaKaRa Prime Script® ET reagent kit Generate cDNA (reverse transcription reaction)
37°C, 15 minutes (reverse transcription reaction)
85°C, 5 seconds (Heat inactivates reverse transcriptase)
4°C, ∞
5. Real-time PCR (SYBR)
(1) Prepare ice and samples (cDNA) (2) Remove controls and test primers from -20°C freezer and place on aluminum foil (3) Remove master mix from -20°C freezer and place on ice (4) Prepare a Micro Amp tube and lid (5) Prepare a light yellow tube for a few minutes of primer and write the name of the primer on the lid (6) Put the master mix and Make a primer mix Master mix 12.5 μL (7.5) × number of samples = μL
H2O , PCR Grade 9μL (5.4) × number of samples = μL
Primer FW 0.5 μL (0.4) × number of samples = μL
Primer RV 0.5 μL (0.4) × number of samples = μL
ROX Reference Dye II 0.5 μL (0.3) × number of samples = μL
* Add the primer to the tube containing the master mix while pipetting.
After mixing well, spin down.
(7) Dispense 23 μL of the prepared Mix into each Micro Amp tube.
(8) After dispensing 23 μL of Mix, add 2 μL of sample to make a total of 25 μL.
(7) 23 μL + (8) 2 μL = 25 μL
(9) Cover the Micro Amp, make sure there are no bubbles and the lid is closed properly, then spin down to drain the liquid from the sides.
(10) Set in a real-time quantitative PCR system (MX3000P (manufactured by Agilent Technologies)).
6. Analyze MX3000P results
1. 脾臓・内臓脂肪・皮下脂肪をマウスから回収
2. 組織をホモジナイズした後、qiagen RNA抽出キットでRNA抽出
3. RNA濃度を測定
4. TaKaRa Prime Script（登録商標） ET reagent kitを使用して cDNAを作製
（逆転写反応）
37℃、 15分 （逆転写反応）
85℃、 5秒 （逆転写酵素を熱失活させる）
4℃、 ∞
5. Real Time PCR (SYBR)
(1) 氷とサンプル(cDNA)を準備する
(2) コントロールと調べるプライマーを-20℃の冷凍庫から出す
アルミホイルの上に置く
(3) マスターMixを -20℃の冷凍庫から出して氷上におく
(4) Micro Ampのtubeと蓋を用意しておく
(5) 薄黄色のtubeをプライマー数分準備し、蓋にプライマーの名前を書いておく
(6) 用意した薄黄色のtubeにマスターMixとプライマーのMixを作る
マスターMix 12.5μL（7.5） × サンプル数 ＝ μL
H2O , PCR Grade 9μL（5.4） × サンプル数 ＝ μL
プライマー FW 0.5μL （0.4） × サンプル数 = μL
プライマー RV 0.5μL （0.4） × サンプル数 = μL
ROX Reference Dye II 0.5μL（0.3） × サンプル数 = μL
※マスターMixを入れたtubeに、プライマーをピペッティングしながら加える。
よく混ぜた後、スピンダウンする。
(7) 作ったMixをMicro Ampのtubeに23μLずつ分注する。
(8) Mix 23μLずつ分注したら、サンプルを2μLずつ加えて、合計25μLにする。
(7) 23μL ＋ (8) 2μL ＝ 25μL
(9) Micro Ampに蓋をして、泡がないか、蓋がきちんとしまっているかを確認したのち、スピンダウンして液を側面から落とす。
(10) リアルタイム定量PCRシステム（MX3000P（アジレント・テクノロジー株式会社製））にセットする。
6. MX3000Pの結果を解析する [qPCR]
1. Harvest spleen, visceral fat, and subcutaneous fat from
37°C, 15 minutes (reverse transcription reaction)
85°C, 5 seconds (Heat inactivates reverse transcriptase)
4°C, ∞
5. Real-time PCR (SYBR)
(1) Prepare ice and samples (cDNA) (2) Remove controls and test primers from -20°C freezer and place on aluminum foil (3) Remove master mix from -20°C freezer and place on ice (4) Prepare a Micro Amp tube and lid (5) Prepare a light yellow tube for a few minutes of primer and write the name of the primer on the lid (6) Put the master mix and Make a primer mix Master mix 12.5 μL (7.5) × number of samples = μL
H2O , PCR Grade 9μL (5.4) × number of samples = μL
Primer FW 0.5 μL (0.4) × number of samples = μL
Primer RV 0.5 μL (0.4) × number of samples = μL
ROX Reference Dye II 0.5 μL (0.3) × number of samples = μL
* Add the primer to the tube containing the master mix while pipetting.
After mixing well, spin down.
(7) Dispense 23 μL of the prepared Mix into each Micro Amp tube.
(8) After dispensing 23 μL of Mix, add 2 μL of sample to make a total of 25 μL.
(7) 23 μL + (8) 2 μL = 25 μL
(9) Cover the Micro Amp, make sure there are no bubbles and the lid is closed properly, then spin down to drain the liquid from the sides.
(10) Set in a real-time quantitative PCR system (MX3000P (manufactured by Agilent Technologies)).
6. Analyze MX3000P results
（Krebs-Hense-leit-HEPES（KRHAG）buffer）
生理食塩水、5% BSA in 生理食塩水、1M KCl、1M CaCl2、1M KH2PO4、1M MgSO4、200mM Hepes、200mM glucose
（コラゲナーゼ）
18mg in 9mL KRHAG （事前に取り分けて-30℃ 使う直前に投入） (Krebs-Hense-leit-HEPES (KRHAG) buffer)
Physiological saline, 5% BSA in physiological saline, 1M KCl, 1M CaCl2, 1M KH2PO4 , 1M MgSO4, 200mM Hepes, 200mM glucose
(collagenase)
18mg in 9mL KRHAG (Available in advance and added just before use at -30℃)
生理食塩水、5% BSA in 生理食塩水、1M KCl、1M CaCl2、1M KH2PO4、1M MgSO4、200mM Hepes、200mM glucose
（コラゲナーゼ）
18mg in 9mL KRHAG （事前に取り分けて-30℃ 使う直前に投入） (Krebs-Hense-leit-HEPES (KRHAG) buffer)
Physiological saline, 5% BSA in physiological saline, 1M KCl, 1M CaCl2, 1M KH2PO4 , 1M MgSO4, 200mM Hepes, 200mM glucose
(collagenase)
18mg in 9mL KRHAG (Available in advance and added just before use at -30℃)
［FACS解析］
(脂肪組織間質血管細胞群（stromal vascular fraction:SVF）の回収)
1. 白色脂肪組織（white adipose tissue：WAT）回収
2. ミンス
3. コラゲナーゼに入れ 37℃ 27回/min
4. 20min 40min の時にタッピング
5. 45min でon ice
6. KRHAG 4mL 追加 ストッキングで濾す
7. 1,500rpm 5min 4℃
8. 上清を吸う。ペレットを2mL残し2mLチューブに移す
9. 1500rpm 5min 4℃
10. 上清を吸って KRHAG 2mL 追加
11. 1,500rpm 5min 4℃
12. 上清を吸って KRHAG 2mL 追加
13. 1,500rpm 5min 4℃
14. 上清を吸って FACS buffer 150μｌ追加
15. lyse Lysis Buffer 1.35mL
16. protect from light 15min 4℃
17. 1,500rpm 5min 4℃
18. 上清 捨てる FACS Buffer 1mL [FACS analysis]
(Recovery of adipose tissue stromal vascular fraction (SVF))
1. Collect white adipose tissue (WAT) 2.Mince 3. Place in collagenase 37°C 27 times/min
4. Tapping at20min 40min 5. On ice at 45min
6. Add 4mL KRHAG Strain through stocking 7. 1,500rpm 5min 4℃
8. Aspirate supernatant. Leave 2mL of the pellet and transfer to a2mL tube 9. 1500rpm 5min 4℃
10. Aspirate the supernatant and add 2 mL of KRHAG
11. 1,500rpm 5min 4℃
12. Aspirate the supernatant and add 2 mL of KRHAG
13. 1,500rpm 5min 4℃
14. Aspirate the supernatant and add 150 μl of FACS buffer
15. Lyse Lysis Buffer 1.35mL
16.protect fromlight 15min 4℃
17. 1,500rpm 5min 4℃
18. Discard supernatant FACS Buffer 1mL
(脂肪組織間質血管細胞群（stromal vascular fraction:SVF）の回収)
1. 白色脂肪組織（white adipose tissue：WAT）回収
2. ミンス
3. コラゲナーゼに入れ 37℃ 27回/min
4. 20min 40min の時にタッピング
5. 45min でon ice
6. KRHAG 4mL 追加 ストッキングで濾す
7. 1,500rpm 5min 4℃
8. 上清を吸う。ペレットを2mL残し2mLチューブに移す
9. 1500rpm 5min 4℃
10. 上清を吸って KRHAG 2mL 追加
11. 1,500rpm 5min 4℃
12. 上清を吸って KRHAG 2mL 追加
13. 1,500rpm 5min 4℃
14. 上清を吸って FACS buffer 150μｌ追加
15. lyse Lysis Buffer 1.35mL
16. protect from light 15min 4℃
17. 1,500rpm 5min 4℃
18. 上清 捨てる FACS Buffer 1mL [FACS analysis]
(Recovery of adipose tissue stromal vascular fraction (SVF))
1. Collect white adipose tissue (WAT) 2.
4. Tapping at
6. Add 4mL KRHAG Strain through stocking 7. 1,500
8. Aspirate supernatant. Leave 2mL of the pellet and transfer to a
10. Aspirate the supernatant and add 2 mL of KRHAG
11. 1,500
12. Aspirate the supernatant and add 2 mL of KRHAG
13. 1,500
14. Aspirate the supernatant and add 150 μl of FACS buffer
15. Lyse Lysis Buffer 1.35mL
16.protect from
17. 1,500
18. Discard supernatant FACS Buffer 1mL
（脾臓細胞の回収）
1. 脾臓 回収
2. ホモジナイズ
3. 1,500rpm 5min 4℃
4. 上清を吸って FACS buffer 150μｌ追加
5. lyse Lysis Buffer 1.35mL
6. protect from light 15min 4℃
7. 1,500rpm 5min 4℃
8. 上清 捨てる FACS Buffer 1mL (Collection of spleen cells)
1.Collect spleen 2. Homogenize 3. 1,500rpm 5min 4℃
4. Aspirate the supernatant and add 150 μl ofFACS buffer 5. Lyse Lysis Buffer 1.35 mL
6.protect fromlight 15min 4℃
7. 1,500rpm 5min 4℃
8. Discard supernatant FACS Buffer 1mL
1. 脾臓 回収
2. ホモジナイズ
3. 1,500rpm 5min 4℃
4. 上清を吸って FACS buffer 150μｌ追加
5. lyse Lysis Buffer 1.35mL
6. protect from light 15min 4℃
7. 1,500rpm 5min 4℃
8. 上清 捨てる FACS Buffer 1mL (Collection of spleen cells)
1.
4. Aspirate the supernatant and add 150 μl of
6.protect from
7. 1,500
8. Discard supernatant FACS Buffer 1mL
（染色）
1. FC Brock：2%FCS/PBS = (1：100)を加えた後15min 4℃
2. CD45-PECy7：2%FCS/PBS = (1：400)
F4/80-APCCy7：2%FCS/PBS = (1：80)
CD11c-PE：2%FCS/PBS = (1：50)
CD206-APC：2%FCS/PBS = (1：80)を加えた後30min 4℃
3. 2%FCS/PBS を200μL加える
1,500rpm 5min 4℃
4. 上清を捨て2%FCS/PBS を200μL加える
5. 1,500rpm 5min 4℃
6. 上清を捨て2% FCS/PBSを200μL加える
7. FACSにて測定（計測の直前に7AAD 1μLを加える） (staining)
1. FC Brock: 15min 4℃ after adding 2%FCS/PBS = (1:100)
2. CD45-PECy7: 2% FCS/PBS = (1:400)
F4/80-APCCy7: 2% FCS/PBS = (1:80)
CD11c-PE: 2% FCS/PBS = (1:50)
CD206-APC: 30min after adding 2%FCS/PBS = (1:80) 4℃
3. Add 200μL of 2%FCS/PBS 1,500rpm 5min 4℃
4. Discard the supernatant and add 200μL of 2%FCS/PBS 5. 1,500rpm 5min 4℃
6. Discard the supernatant and add 200 μL of 2% FCS/PBS 7. Measure with FACS (add 1 μL of 7AAD just before measurement)
1. FC Brock：2%FCS/PBS = (1：100)を加えた後15min 4℃
2. CD45-PECy7：2%FCS/PBS = (1：400)
F4/80-APCCy7：2%FCS/PBS = (1：80)
CD11c-PE：2%FCS/PBS = (1：50)
CD206-APC：2%FCS/PBS = (1：80)を加えた後30min 4℃
3. 2%FCS/PBS を200μL加える
1,500rpm 5min 4℃
4. 上清を捨て2%FCS/PBS を200μL加える
5. 1,500rpm 5min 4℃
6. 上清を捨て2% FCS/PBSを200μL加える
7. FACSにて測定（計測の直前に7AAD 1μLを加える） (staining)
1. FC Brock: 15min 4℃ after adding 2%FCS/PBS = (1:100)
2. CD45-PECy7: 2% FCS/PBS = (1:400)
F4/80-APCCy7: 2% FCS/PBS = (1:80)
CD11c-PE: 2% FCS/PBS = (1:50)
CD206-APC: 30min after adding 2%FCS/PBS = (1:80) 4℃
3. Add 200μL of 2%FCS/PBS 1,500
4. Discard the supernatant and add 200μL of 2%FCS/
6. Discard the supernatant and add 200 μL of 2% FCS/PBS 7. Measure with FACS (add 1 μL of 7AAD just before measurement)
＜実施例５：抗CD206抗体の代謝障害改善作用の評価＞
以下のようにして、本発明の抗CD206抗体の代謝障害改善作用を評価した。 <Example 5: Evaluation of metabolic disorder improving effect of anti-CD206 antibody>
The anti-CD206 antibody of the present invention was evaluated for its ameliorating effect on metabolic disorders as follows.
以下のようにして、本発明の抗CD206抗体の代謝障害改善作用を評価した。 <Example 5: Evaluation of metabolic disorder improving effect of anti-CD206 antibody>
The anti-CD206 antibody of the present invention was evaluated for its ameliorating effect on metabolic disorders as follows.
（高脂肪食負荷マウス）
C57BL/6Jマウス（日本クレア株式会社製)に、8週齢から16週間、高脂肪食（RESERCH DIETS社製 超高脂肪飼料60kcal%脂肪含有量、ラード使用）を負荷した、肥満の糖尿病のモデルマウスを作製した。 (high-fat diet-loaded mice)
C57BL/6J mice (manufactured by CLEA Japan, Inc.) were fed a high-fat diet (reserch diets ultra-high-fat diet containing 60 kcal% fat and lard) from 8 weeks of age to 16 weeks as an obese diabetic model. A mouse was produced.
C57BL/6Jマウス（日本クレア株式会社製)に、8週齢から16週間、高脂肪食（RESERCH DIETS社製 超高脂肪飼料60kcal%脂肪含有量、ラード使用）を負荷した、肥満の糖尿病のモデルマウスを作製した。 (high-fat diet-loaded mice)
C57BL/6J mice (manufactured by CLEA Japan, Inc.) were fed a high-fat diet (reserch diets ultra-high-fat diet containing 60 kcal% fat and lard) from 8 weeks of age to 16 weeks as an obese diabetic model. A mouse was produced.
［抗体投与］
PBSに希釈した被験抗体および対照としてPBSを、それぞれ1μg/マウス体重(g)の濃度で0,48,96時間の計3回腹腔内に投与した（No. 83 n=4）。投与4日（96時間）後、実験を行った。 [Antibody administration]
A test antibody diluted in PBS and PBS as a control were each intraperitoneally administered at a concentration of 1 μg/mouse body weight (g) three times at 0, 48, and 96 hours (No. 83 n=4). Experiments were performed 4 days (96 hours) after administration.
PBSに希釈した被験抗体および対照としてPBSを、それぞれ1μg/マウス体重(g)の濃度で0,48,96時間の計3回腹腔内に投与した（No. 83 n=4）。投与4日（96時間）後、実験を行った。 [Antibody administration]
A test antibody diluted in PBS and PBS as a control were each intraperitoneally administered at a concentration of 1 μg/mouse body weight (g) three times at 0, 48, and 96 hours (No. 83 n=4). Experiments were performed 4 days (96 hours) after administration.
［糖負荷試験］
1. 実験開始4時間前に餌を抜き、ケージを交換した
2. 1g/kgのブドウ糖をマウスの腹腔内に注射投与した
3. ブドウ糖投与前と投与後15,30,60,90,120分に尾静脈より採血した
4. 簡易血糖測定器にて血糖を測定した [Sugar tolerance test]
1. Food was withdrawn 4 hours before the start of the experiment and the cage was changed 2. 1g/kg glucose was injected intraperitoneally into themice 3. Tails were taken before and 15, 30, 60, 90 and 120 minutes after administration of glucose Blood was collected from a vein 4. Blood glucose was measured with a simple blood glucose meter
1. 実験開始4時間前に餌を抜き、ケージを交換した
2. 1g/kgのブドウ糖をマウスの腹腔内に注射投与した
3. ブドウ糖投与前と投与後15,30,60,90,120分に尾静脈より採血した
4. 簡易血糖測定器にて血糖を測定した [Sugar tolerance test]
1. Food was withdrawn 4 hours before the start of the experiment and the cage was changed 2. 1g/kg glucose was injected intraperitoneally into the
［インスリン負荷試験］
1. 実験開始4時間前に餌を抜き、ケージを交換した
2. 1単位/kgのインスリンをマウスの腹腔内に注射投与した
3. インスリン投与前と投与後15,30,45,60,90,120分に尾静脈より採血した
4. 簡易血糖測定器にて血糖を測定した [Insulin tolerance test]
1. Food was withdrawn 4 hours before the start of the experiment and the cage was changed 2. 1 unit/kg of insulin was injected intraperitoneally into themice 3. Before and after insulin administration15,30,45,60,90,120 4. Blood glucose was measured with a simple blood glucose meter.
1. 実験開始4時間前に餌を抜き、ケージを交換した
2. 1単位/kgのインスリンをマウスの腹腔内に注射投与した
3. インスリン投与前と投与後15,30,45,60,90,120分に尾静脈より採血した
4. 簡易血糖測定器にて血糖を測定した [Insulin tolerance test]
1. Food was withdrawn 4 hours before the start of the experiment and the cage was changed 2. 1 unit/kg of insulin was injected intraperitoneally into the
結果を図６に示す。本発明で作製した抗CD206抗体を投与したマウスでは、糖負荷試験の結果より、耐糖機能障害を改善することが示された。また、インスリン負荷試験の結果より、インスリン抵抗性を改善することが示された。したがって、本発明の抗CD206抗体は、代謝障害の予防または治療用医薬組成物として用いることができると考えられる。
The results are shown in Figure 6. The results of the glucose tolerance test showed that the anti-CD206 antibody produced in the present invention was administered to mice to improve impaired glucose tolerance. In addition, the results of insulin tolerance test showed that it improved insulin resistance. Therefore, it is believed that the anti-CD206 antibodies of the present invention can be used as pharmaceutical compositions for preventing or treating metabolic disorders.
＜実施例６：抗CD206抗体の2型糖尿病モデル動物での有効性の検討＞
肥満2型糖尿病モデルマウスへ濃度の異なる抗CD206抗体を皮下投与し、効果と効果の継続期間を試験した。
実施例５において作製した高脂肪食負荷マウスを2型糖尿病モデル動物として用いた。
PBSに希釈した被験抗体（No. 83 n=5～6）0.3, 1, 3, 10, 30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=5）0.3, 1, 3, 10, 30μg/マウス体重(g)を、それぞれ皮下に投与し、投与7, 10, 14日後に、実施例５と同様にして、糖負荷試験を行った。 <Example 6: Examination of efficacy of anti-CD206 antibody intype 2 diabetes model animals>
Different concentrations of anti-CD206 antibodies were subcutaneously administered toobese type 2 diabetes model mice, and the effect and duration of the effect were examined.
The high-fat diet-fed mice prepared in Example 5 were used astype 2 diabetes model animals.
Test antibody diluted in PBS (No. 83 n=5-6) 0.3, 1, 3, 10, 30 μg/mouse body weight (g) and test antibody diluted in PBS (Hel2 n=5) 0.3, 1, 3, 10, 30 μg/mouse body weight (g) was administered subcutaneously, and glucose tolerance test was performed in the same manner as in Example 5 7, 10, 14 days after administration.
肥満2型糖尿病モデルマウスへ濃度の異なる抗CD206抗体を皮下投与し、効果と効果の継続期間を試験した。
実施例５において作製した高脂肪食負荷マウスを2型糖尿病モデル動物として用いた。
PBSに希釈した被験抗体（No. 83 n=5～6）0.3, 1, 3, 10, 30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=5）0.3, 1, 3, 10, 30μg/マウス体重(g)を、それぞれ皮下に投与し、投与7, 10, 14日後に、実施例５と同様にして、糖負荷試験を行った。 <Example 6: Examination of efficacy of anti-CD206 antibody in
Different concentrations of anti-CD206 antibodies were subcutaneously administered to
The high-fat diet-fed mice prepared in Example 5 were used as
Test antibody diluted in PBS (No. 83 n=5-6) 0.3, 1, 3, 10, 30 μg/mouse body weight (g) and test antibody diluted in PBS (Hel2 n=5) 0.3, 1, 3, 10, 30 μg/mouse body weight (g) was administered subcutaneously, and glucose tolerance test was performed in the same manner as in Example 5 7, 10, 14 days after administration.
異なる濃度の被験抗体による結果を図７に、被験抗体および対象抗体の比較結果を図８に示す。本発明で作製した抗CD206抗体（30μg/BW）を皮下に単回投与することで、1週間耐糖能の改善効果が確認できた。
Figure 7 shows the results with different concentrations of the test antibody, and Figure 8 shows the comparison results of the test antibody and the target antibody. A single subcutaneous administration of the anti-CD206 antibody (30 μg/BW) prepared according to the present invention was confirmed to improve glucose tolerance for one week.
＜実施例７：抗CD206抗体の代謝障害改善作用の作用機序の解明＞
（CD206ノックアウトマウス）
CD206ノックアウトマウスはThe Jackson Laboratoryより購入した。なお、同腹子マウスをWTマウスとした。
これらを用いて、実施例５と同様に高脂肪食負荷を行った。 <Example 7: Elucidation of mechanism of action of anti-CD206 antibody to improve metabolic disorders>
(CD206 knockout mouse)
CD206 knockout mice were purchased from The Jackson Laboratory. The littermate mice were WT mice.
Using these, high-fat diet loading was performed in the same manner as in Example 5.
（CD206ノックアウトマウス）
CD206ノックアウトマウスはThe Jackson Laboratoryより購入した。なお、同腹子マウスをWTマウスとした。
これらを用いて、実施例５と同様に高脂肪食負荷を行った。 <Example 7: Elucidation of mechanism of action of anti-CD206 antibody to improve metabolic disorders>
(CD206 knockout mouse)
CD206 knockout mice were purchased from The Jackson Laboratory. The littermate mice were WT mice.
Using these, high-fat diet loading was performed in the same manner as in Example 5.
［糖負荷試験およびインスリン負荷試験］
WTマウスへPBSに希釈した被験抗体（No. 83 n=3）30μg/マウス体重(g)を、CD206KOマウスへPBSに希釈した被験抗体（No. 83 n=3）30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=3）30μg/マウス体重(g)を、それぞれ皮下に投与し、投与7日後に、実施例５と同様にして、糖負荷試験およびインスリン負荷試験を行った。 [Glucose tolerance test and insulin tolerance test]
Test antibody (No. 83 n=3) 30 μg/mouse weight (g) diluted in PBS to WT mice, Test antibody (No. 83 n=3) diluted in PBS toCD206KO mice 30 μg/mouse weight (g) and 30 μg/mouse body weight (g) of the target antibody (Hel2 n=3) diluted in PBS were administered subcutaneously, respectively, and 7 days after administration, glucose tolerance test and insulin tolerance test were performed in the same manner as in Example 5. rice field.
WTマウスへPBSに希釈した被験抗体（No. 83 n=3）30μg/マウス体重(g)を、CD206KOマウスへPBSに希釈した被験抗体（No. 83 n=3）30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=3）30μg/マウス体重(g)を、それぞれ皮下に投与し、投与7日後に、実施例５と同様にして、糖負荷試験およびインスリン負荷試験を行った。 [Glucose tolerance test and insulin tolerance test]
Test antibody (No. 83 n=3) 30 μg/mouse weight (g) diluted in PBS to WT mice, Test antibody (No. 83 n=3) diluted in PBS to
WTマウスへ被験抗体を、CD206KOマウスに被験抗体または対象抗体を投与した、各マウスの抗体投与前と抗体投与1週間後の血糖値の改善率（CD206KOマウスに対照抗体を投与した群を1として計算）を図９に示す。CD206KOマウスに被験抗体または対象抗体を投与したマウスでは有意な耐糖機能障害およびインスリン抵抗性の改善は見られなかった。一方、WTマウスへ被験抗体を投与した群では耐糖機能障害およびインスリン抵抗性の改善が認められた。これらより、抗CD206抗体の効果はCD206を介した効果であることが証明された。
The rate of improvement in blood glucose levels before antibody administration and one week after antibody administration in each mouse after administering the test antibody to WT mice and the test antibody or target antibody to CD206KO mice (the group administered the control antibody to CD206KO mice is regarded as 1) calculation) is shown in FIG. No significant improvement in glucose tolerance or insulin resistance was observed in CD206KO mice administered the test or subject antibodies. On the other hand, improvement in glucose tolerance dysfunction and insulin resistance was observed in the group administered the test antibody to WT mice. These results demonstrated that the effect of the anti-CD206 antibody was mediated by CD206.
＜実施例８：抗CD206抗体の初期の安全性の確認＞
肥満2型糖尿病モデルマウスへ抗CD206抗体を皮下投与し、安全性を試験した。
実施例５において作製した高脂肪食負荷マウスを2型糖尿病モデル動物として用いた。
PBSに希釈した被験抗体（No. 83 n=11）30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=10）30μg/マウス体重(g)を、それぞれ週1回計4回皮下に投与し、28日間の体重の変化および最終週の摂餌量を確認した。また、28日後の肝臓（Liver）・内臓脂肪（eWAT）・鼠径部皮下脂肪（iWAT）をマウスから回収し、組織重量を確認した。 <Example 8: Confirmation of initial safety of anti-CD206 antibody>
Anti-CD206 antibody was subcutaneously administered toobese type 2 diabetes model mice, and safety was tested.
The high-fat diet-fed mice prepared in Example 5 were used astype 2 diabetes model animals.
Test antibody (No. 83 n=11) diluted in PBS at 30 μg/mouse weight (g) and target antibody (Hel2 n=10) diluted in PBS at 30 μg/mouse weight (g) each once a week for a total of 4 times After subcutaneous administration, changes in body weight for 28 days and food intake in the final week were confirmed. Twenty-eight days later, liver (Liver), visceral fat (eWAT), and inguinal subcutaneous fat (iWAT) were collected from the mice, and tissue weights were confirmed.
肥満2型糖尿病モデルマウスへ抗CD206抗体を皮下投与し、安全性を試験した。
実施例５において作製した高脂肪食負荷マウスを2型糖尿病モデル動物として用いた。
PBSに希釈した被験抗体（No. 83 n=11）30μg/マウス体重(g)およびPBSに希釈した対象抗体（Hel2 n=10）30μg/マウス体重(g)を、それぞれ週1回計4回皮下に投与し、28日間の体重の変化および最終週の摂餌量を確認した。また、28日後の肝臓（Liver）・内臓脂肪（eWAT）・鼠径部皮下脂肪（iWAT）をマウスから回収し、組織重量を確認した。 <Example 8: Confirmation of initial safety of anti-CD206 antibody>
Anti-CD206 antibody was subcutaneously administered to
The high-fat diet-fed mice prepared in Example 5 were used as
Test antibody (No. 83 n=11) diluted in PBS at 30 μg/mouse weight (g) and target antibody (Hel2 n=10) diluted in PBS at 30 μg/mouse weight (g) each once a week for a total of 4 times After subcutaneous administration, changes in body weight for 28 days and food intake in the final week were confirmed. Twenty-eight days later, liver (Liver), visceral fat (eWAT), and inguinal subcutaneous fat (iWAT) were collected from the mice, and tissue weights were confirmed.
体重、摂餌量の結果を図１０に、組織重量の結果を図１１に示す。抗CD206抗体の効果が28日間持続するよう投与を続けたが、突然死や体重の減少、摂餌量の減少、異常行動は見られなかった。
なお、皮下脂肪の縮小が確認された。その他、組織所見に異常は見られなかった。 The results of body weight and food intake are shown in FIG. 10, and the results of tissue weight are shown in FIG. Administration of the anti-CD206 antibody continued for 28 days, but no sudden death, weight loss, decreased food consumption, or abnormal behavior was observed.
A reduction in subcutaneous fat was confirmed. No other abnormal tissue findings were observed.
なお、皮下脂肪の縮小が確認された。その他、組織所見に異常は見られなかった。 The results of body weight and food intake are shown in FIG. 10, and the results of tissue weight are shown in FIG. Administration of the anti-CD206 antibody continued for 28 days, but no sudden death, weight loss, decreased food consumption, or abnormal behavior was observed.
A reduction in subcutaneous fat was confirmed. No other abnormal tissue findings were observed.
本発明の抗CD206抗体またはその機能的断片は、耐糖機能障害、インスリン抵抗性および糖尿病等の予防または治療用医薬組成物として用いることができる。
The anti-CD206 antibody or functional fragment thereof of the present invention can be used as a pharmaceutical composition for prevention or treatment of impaired glucose tolerance, insulin resistance, diabetes, and the like.
＜配列表の説明＞
配列番号１：抗CD206抗体（No.10）の重鎖可変領域のアミノ酸配列
配列番号２：抗CD206抗体（No.10）の軽鎖可変領域のアミノ酸配列
配列番号３：抗CD206抗体（No.11）の重鎖可変領域のアミノ酸配列
配列番号４：抗CD206抗体（No.11）の軽鎖可変領域のアミノ酸配列
配列番号５：抗CD206抗体（No.27）の重鎖可変領域のアミノ酸配列
配列番号６：抗CD206抗体（No.27）の軽鎖可変領域のアミノ酸配列
配列番号７：抗CD206抗体（No.35）の重鎖可変領域のアミノ酸配列
配列番号８：抗CD206抗体（No.35）の軽鎖可変領域のアミノ酸配列
配列番号９：抗CD206抗体（No.40）の重鎖可変領域のアミノ酸配列
配列番号１０：抗CD206抗体（No.40）の軽鎖可変領域のアミノ酸配列
配列番号１１：抗CD206抗体（No.44）の重鎖可変領域のアミノ酸配列
配列番号１２：抗CD206抗体（No.44）の軽鎖可変領域のアミノ酸配列
配列番号１３：抗CD206抗体（No.47）の重鎖可変領域のアミノ酸配列
配列番号１４：抗CD206抗体（No.47）の軽鎖可変領域のアミノ酸配列
配列番号１５：抗CD206抗体（No.54）の重鎖可変領域のアミノ酸配列
配列番号１６：抗CD206抗体（No.54）の軽鎖可変領域のアミノ酸配列
配列番号１７：抗CD206抗体（No.59）の重鎖可変領域のアミノ酸配列
配列番号１８：抗CD206抗体（No.59）の軽鎖可変領域のアミノ酸配列
配列番号１９：抗CD206抗体（No.64）の重鎖可変領域のアミノ酸配列
配列番号２０：抗CD206抗体（No.64）の軽鎖可変領域のアミノ酸配列
配列番号２１：抗CD206抗体（No.72）の重鎖可変領域のアミノ酸配列
配列番号２２：抗CD206抗体（No.72）の軽鎖可変領域のアミノ酸配列
配列番号２３：抗CD206抗体（No.73）の重鎖可変領域のアミノ酸配列
配列番号２４：抗CD206抗体（No.73）の軽鎖可変領域のアミノ酸配列
配列番号２５：抗CD206抗体（No.83）の重鎖可変領域のアミノ酸配列
配列番号２６：抗CD206抗体（No.83）の軽鎖可変領域のアミノ酸配列
配列番号２７：抗CD206抗体（No.10）のHCDR1のアミノ酸配列
配列番号２８：抗CD206抗体（No.10）のHCDR2のアミノ酸配列
配列番号２９：抗CD206抗体（No.10）のHCDR3のアミノ酸配列
配列番号３０：抗CD206抗体（No.10）のLCDR1のアミノ酸配列
配列番号３１：抗CD206抗体（No.10）のLCDR3のアミノ酸配列
配列番号３２：抗CD206抗体（No.11）のHCDR1のアミノ酸配列
配列番号３３：抗CD206抗体（No.11）のHCDR2のアミノ酸配列
配列番号３４：抗CD206抗体（No.11）のHCDR3のアミノ酸配列
配列番号３５：抗CD206抗体（No.11）のLCDR1のアミノ酸配列
配列番号３６：抗CD206抗体（No.11）のLCDR3のアミノ酸配列
配列番号３７：抗CD206抗体（No.27）のHCDR1のアミノ酸配列
配列番号３８：抗CD206抗体（No.27）のHCDR2のアミノ酸配列
配列番号３９：抗CD206抗体（No.27）のHCDR3のアミノ酸配列
配列番号４０：抗CD206抗体（No.27）のLCDR1のアミノ酸配列
配列番号４１：抗CD206抗体（No.27）のLCDR3のアミノ酸配列
配列番号４２：抗CD206抗体（No.35）のHCDR1のアミノ酸配列
配列番号４３：抗CD206抗体（No.35）のHCDR2のアミノ酸配列
配列番号４４：抗CD206抗体（No.35）のHCDR3のアミノ酸配列
配列番号４５：抗CD206抗体（No.35）のLCDR1のアミノ酸配列
配列番号４６：抗CD206抗体（No.35）のLCDR3のアミノ酸配列
配列番号４７：抗CD206抗体（No.40）のHCDR1のアミノ酸配列
配列番号４８：抗CD206抗体（No.40）のHCDR2のアミノ酸配列
配列番号４９：抗CD206抗体（No.40）のHCDR3のアミノ酸配列
配列番号５０：抗CD206抗体（No.40）のLCDR1のアミノ酸配列
配列番号５１：抗CD206抗体（No.40）のLCDR3のアミノ酸配列
配列番号５２：抗CD206抗体（No.44）のHCDR1のアミノ酸配列
配列番号５３：抗CD206抗体（No.44）のHCDR2のアミノ酸配列
配列番号５４：抗CD206抗体（No.44）のHCDR3のアミノ酸配列
配列番号５５：抗CD206抗体（No.44）のLCDR1のアミノ酸配列
配列番号５６：抗CD206抗体（No.44）のLCDR3のアミノ酸配列
配列番号５７：抗CD206抗体（No.47）のHCDR1のアミノ酸配列
配列番号５８：抗CD206抗体（No.47）のHCDR2のアミノ酸配列
配列番号５９：抗CD206抗体（No.47）のHCDR3のアミノ酸配列
配列番号６０：抗CD206抗体（No.47）のLCDR1のアミノ酸配列
配列番号６１：抗CD206抗体（No.47）のLCDR3のアミノ酸配列
配列番号６２：抗CD206抗体（No.54）のHCDR1のアミノ酸配列
配列番号６３：抗CD206抗体（No.54）のHCDR2のアミノ酸配列
配列番号６４：抗CD206抗体（No.54）のHCDR3のアミノ酸配列
配列番号６５：抗CD206抗体（No.54）のLCDR1のアミノ酸配列
配列番号６６：抗CD206抗体（No.54）のLCDR3のアミノ酸配列
配列番号６７：抗CD206抗体（No.59）のHCDR1のアミノ酸配列
配列番号６８：抗CD206抗体（No.59）のHCDR2のアミノ酸配列
配列番号６９：抗CD206抗体（No.59）のHCDR3のアミノ酸配列
配列番号７０：抗CD206抗体（No.59）のLCDR1のアミノ酸配列
配列番号７１：抗CD206抗体（No.59）のLCDR3のアミノ酸配列
配列番号７２：抗CD206抗体（No.64）のHCDR1のアミノ酸配列
配列番号７３：抗CD206抗体（No.64）のHCDR2のアミノ酸配列
配列番号７４：抗CD206抗体（No.64）のHCDR3のアミノ酸配列
配列番号７５：抗CD206抗体（No.64）のLCDR1のアミノ酸配列
配列番号７６：抗CD206抗体（No.64）のLCDR3のアミノ酸配列
配列番号７７：抗CD206抗体（No.72）のHCDR1のアミノ酸配列
配列番号７８：抗CD206抗体（No.72）のHCDR2のアミノ酸配列
配列番号７９：抗CD206抗体（No.72）のHCDR3のアミノ酸配列
配列番号８０：抗CD206抗体（No.72）のLCDR1のアミノ酸配列
配列番号８１：抗CD206抗体（No.72）のLCDR3のアミノ酸配列
配列番号８２：抗CD206抗体（No.73）のHCDR1のアミノ酸配列
配列番号８３：抗CD206抗体（No.73）のHCDR2のアミノ酸配列
配列番号８４：抗CD206抗体（No.73）のHCDR3のアミノ酸配列
配列番号８５：抗CD206抗体（No.73）のLCDR1のアミノ酸配列
配列番号８６：抗CD206抗体（No.73）のLCDR3のアミノ酸配列
配列番号８７：抗CD206抗体（No.83）のHCDR1のアミノ酸配列
配列番号８８：抗CD206抗体（No.83）のHCDR2のアミノ酸配列
配列番号８９：抗CD206抗体（No.83）のHCDR3のアミノ酸配列
配列番号９０：抗CD206抗体（No.83）のLCDR1のアミノ酸配列
配列番号９１：抗CD206抗体（No.83）のLCDR3のアミノ酸配列
配列番９２：抗CD206抗体（No.10）の重鎖可変領域をコードする塩基配列
配列番号９３：抗CD206抗体（No.10）の軽鎖可変領域をコードする塩基配列
配列番号９４：抗CD206抗体（No.11）の重鎖可変領域をコードする塩基配列
配列番号９５：抗CD206抗体（No.11）の軽鎖可変領域をコードする塩基配列
配列番号９６：抗CD206抗体（No.27）の重鎖可変領域をコードする塩基配列
配列番号９７：抗CD206抗体（No.27）の軽鎖可変領域をコードする塩基配列
配列番号９８：抗CD206抗体（No.35）の重鎖可変領域をコードする塩基配列
配列番号９９：抗CD206抗体（No.35）の軽鎖可変領域をコードする塩基配列
配列番号１００：抗CD206抗体（No.40）の重鎖可変領域をコードする塩基配列
配列番号１０１：抗CD206抗体（No.40）の軽鎖可変領域をコードする塩基配列
配列番号１０２：抗CD206抗体（No.44）の重鎖可変領域をコードする塩基配列
配列番号１０３：抗CD206抗体（No.44）の軽鎖可変領域をコードする塩基配列
配列番号１０４：抗CD206抗体（No.47）の重鎖可変領域をコードする塩基配列
配列番号１０５：抗CD206抗体（No.47）の軽鎖可変領域をコードする塩基配列
配列番号１０６：抗CD206抗体（No.54）の重鎖可変領域をコードする塩基配列
配列番号１０７：抗CD206抗体（No.54）の軽鎖可変領域をコードする塩基配列
配列番号１０８：抗CD206抗体（No.59）の重鎖可変領域をコードする塩基配列
配列番号１０９：抗CD206抗体（No.59）の軽鎖可変領域をコードする塩基配列
配列番号１１０：抗CD206抗体（No.64）の重鎖可変領域をコードする塩基配列
配列番号１１１：抗CD206抗体（No.64）の軽鎖可変領域をコードする塩基配列
配列番号１１２：抗CD206抗体（No.72）の重鎖可変領域をコードする塩基配列
配列番号１１３：抗CD206抗体（No.72）の軽鎖可変領域をコードする塩基配列
配列番号１１４：抗CD206抗体（No.73）の重鎖可変領域をコードする塩基配列
配列番号１１５：抗CD206抗体（No.73）の軽鎖可変領域をコードする塩基配列
配列番号１１６：抗CD206抗体（No.83）の重鎖可変領域をコードする塩基配列
配列番号１１７：抗CD206抗体（No.83）の軽鎖可変領域をコードする塩基配列
配列番号１１８：マウスCD206コーディング領域の塩基配列 <Description of Sequence Listing>
SEQ ID NO: 1: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 2: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 3: Anti-CD206 antibody (No. 11) heavy chain variable region amino acid sequence SEQ ID NO: 4: light chain variable region amino acid sequence of anti-CD206 antibody (No. 11) SEQ ID NO: 5: heavy chain variable region amino acid sequence of anti-CD206 antibody (No. 27) SEQ ID NO: 6: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 27) SEQ ID NO: 7: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 35) SEQ ID NO: 8: Anti-CD206 antibody (No. 35) SEQ ID NO:9: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No.40) SEQ ID NO:10: Amino acid sequence of light chain variable region of anti-CD206 antibody (No.40) SEQ ID NO: 11: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 12: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 13: Anti-CD206 antibody (No. 47) SEQ ID NO: 14: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 15: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 16: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 17: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 59) SEQ ID NO: 18: Anti-CD206 antibody (No. 59) SEQ ID NO: 19: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 20: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 21: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 72) SEQ ID NO: 22: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 72) SEQ ID NO: 23: Anti-CD206 antibody (No. 73) SEQ ID NO: 24: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 73) SEQ ID NO: 25: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 83) SEQ ID NO: 26: Amino acid sequence of light chain variable region of anti-CD206 antibody (No.83) SEQ ID NO: 27: Amino acid sequence of HCDR1 of anti-CD206 antibody (No. 10) SEQ ID NO: 28: Anti-CD2 Amino acid sequence of HCDR2 of 06 antibody (No.10) SEQ ID NO:29: Amino acid sequence of HCDR3 of anti-CD206 antibody (No.10) SEQ ID NO:30: Amino acid sequence of LCDR1 of anti-CD206 antibody (No.10) SEQ ID NO:31: LCDR3 amino acid sequence of anti-CD206 antibody (No.10) SEQ ID NO:32: HCDR1 amino acid sequence of anti-CD206 antibody (No.11) SEQ ID NO:33: HCDR2 amino acid sequence of anti-CD206 antibody (No.11) SEQ ID NO:34 : amino acid sequence of HCDR3 of anti-CD206 antibody (No.11) SEQ ID NO:35: amino acid sequence of LCDR1 of anti-CD206 antibody (No.11) SEQ ID NO:36: amino acid sequence of LCDR3 of anti-CD206 antibody (No.11) SEQ ID NO: 37: HCDR1 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO:38: HCDR2 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO:39: HCDR3 amino acid sequence of anti-CD206 antibody (No.27) No. 40: LCDR1 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO: 41: LCDR3 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO: 42: HCDR1 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 43: HCDR2 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 44: HCDR3 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 45: LCDR1 amino acid of anti-CD206 antibody (No.35) Sequences SEQ ID NO: 46: LCDR3 amino acid sequence of anti-CD206 antibody (No. 35) SEQ ID NO: 47: HCDR1 amino acid sequence of anti-CD206 antibody (No. 40) SEQ ID NO: 48: HCDR2 of anti-CD206 antibody (No. 40) Amino acid sequences SEQ ID NO: 49: Amino acid sequence of HCDR3 of anti-CD206 antibody (No. 40) SEQ ID NO: 50: Amino acid sequence of LCDR1 of anti-CD206 antibody (No. 40) SEQ ID NO: 51: LCDR3 of anti-CD206 antibody (No. 40) SEQ ID NO: 52: HCDR1 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 53: HCDR2 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 54: anti-CD206 antibody (No.44) HCDR3 amino acid sequence SEQ ID NO: 55: LCDR1 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: No. 56: LCDR3 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 57: HCDR1 amino acid sequence of anti-CD206 antibody (No.47) SEQ ID NO: 58: HCDR2 amino acid sequence of anti-CD206 antibody (No.47) SEQ ID NO: 59: HCDR3 amino acid sequence of anti-CD206 antibody (No. 47) SEQ ID NO: 60: LCDR1 amino acid sequence of anti-CD206 antibody (No. 47) SEQ ID NO: 61: LCDR3 amino acid of anti-CD206 antibody (No. 47) Sequences SEQ ID NO:62: HCDR1 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO:63: HCDR2 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO:64: HCDR3 of anti-CD206 antibody (No.54) Amino acid sequences SEQ ID NO: 65: LCDR1 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO: 66: LCDR3 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO: 67: HCDR1 of anti-CD206 antibody (No.59) SEQ ID NO: 68: HCDR2 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO: 69: HCDR3 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO: 70: anti-CD206 antibody (No.59) LCDR1 amino acid sequence SEQ ID NO:71: LCDR3 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO:72: HCDR1 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:73: anti-CD206 antibody (No.64) SEQ ID NO:74: HCDR3 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:75: LCDR1 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:76: anti-CD206 antibody (No.64 ) SEQ ID NO: 77: HCDR1 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 78: HCDR2 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 79: anti-CD206 antibody (No. 72) HCDR3 amino acid sequence SEQ ID NO: 80: LCDR1 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 81: LCDR3 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 82: anti-CD206 antibody (No. 73) HCDR1 amino acid sequence SEQ ID NO:83: HCDR2 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 84: HCDR3 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 85: LCDR1 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 86: anti-CD206 antibody (No.73) LCDR3 amino acid sequence SEQ ID NO:87: HCDR1 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:88: HCDR2 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:89: anti-CD206 antibody (No.83) SEQ ID NO:90: LCDR1 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:91: LCDR3 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:92: anti-CD206 antibody (No.10 ) SEQ ID NO: 93: Nucleotide sequence encoding the light chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 94: Heavy chain variable region of anti-CD206 antibody (No. 11) SEQ ID NO:95: Nucleotide sequence encoding the light chain variable region of anti-CD206 antibody (No.11) SEQ ID NO:96: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.27) No. 97: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 27) SEQ ID NO: 98: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 35) SEQ ID NO: 99: Anti-CD206 antibody Nucleotide sequence encoding the light chain variable region of (No.35) SEQ ID NO: 100: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.40) SEQ ID NO: 101: Anti-CD206 antibody (No.40) Nucleotide sequence encoding light chain variable region SEQ ID NO: 102: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 103: Encoding light chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 104: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 105: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 106 : Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.54) SEQ ID NO: 107: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 108: Anti-CD206 antibody (No. 59) SEQ ID NO: 109: anti-CD206 antibody (No 59) SEQ ID NO: 110: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 111: Light chain of anti-CD206 antibody (No. 64) Nucleotide sequence encoding variable region SEQ ID NO: 112: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.72) SEQ ID NO: 113: Nucleotide encoding light chain variable region of anti-CD206 antibody (No.72) Sequence SEQ ID NO: 114: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.73) SEQ ID NO: 115: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No.73) SEQ ID NO: 116: Anti Nucleotide sequence encoding heavy chain variable region of CD206 antibody (No.83) SEQ ID NO: 117: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 83) SEQ ID NO: 118: Nucleotide sequence of mouse CD206 coding region
配列番号１：抗CD206抗体（No.10）の重鎖可変領域のアミノ酸配列
配列番号２：抗CD206抗体（No.10）の軽鎖可変領域のアミノ酸配列
配列番号３：抗CD206抗体（No.11）の重鎖可変領域のアミノ酸配列
配列番号４：抗CD206抗体（No.11）の軽鎖可変領域のアミノ酸配列
配列番号５：抗CD206抗体（No.27）の重鎖可変領域のアミノ酸配列
配列番号６：抗CD206抗体（No.27）の軽鎖可変領域のアミノ酸配列
配列番号７：抗CD206抗体（No.35）の重鎖可変領域のアミノ酸配列
配列番号８：抗CD206抗体（No.35）の軽鎖可変領域のアミノ酸配列
配列番号９：抗CD206抗体（No.40）の重鎖可変領域のアミノ酸配列
配列番号１０：抗CD206抗体（No.40）の軽鎖可変領域のアミノ酸配列
配列番号１１：抗CD206抗体（No.44）の重鎖可変領域のアミノ酸配列
配列番号１２：抗CD206抗体（No.44）の軽鎖可変領域のアミノ酸配列
配列番号１３：抗CD206抗体（No.47）の重鎖可変領域のアミノ酸配列
配列番号１４：抗CD206抗体（No.47）の軽鎖可変領域のアミノ酸配列
配列番号１５：抗CD206抗体（No.54）の重鎖可変領域のアミノ酸配列
配列番号１６：抗CD206抗体（No.54）の軽鎖可変領域のアミノ酸配列
配列番号１７：抗CD206抗体（No.59）の重鎖可変領域のアミノ酸配列
配列番号１８：抗CD206抗体（No.59）の軽鎖可変領域のアミノ酸配列
配列番号１９：抗CD206抗体（No.64）の重鎖可変領域のアミノ酸配列
配列番号２０：抗CD206抗体（No.64）の軽鎖可変領域のアミノ酸配列
配列番号２１：抗CD206抗体（No.72）の重鎖可変領域のアミノ酸配列
配列番号２２：抗CD206抗体（No.72）の軽鎖可変領域のアミノ酸配列
配列番号２３：抗CD206抗体（No.73）の重鎖可変領域のアミノ酸配列
配列番号２４：抗CD206抗体（No.73）の軽鎖可変領域のアミノ酸配列
配列番号２５：抗CD206抗体（No.83）の重鎖可変領域のアミノ酸配列
配列番号２６：抗CD206抗体（No.83）の軽鎖可変領域のアミノ酸配列
配列番号２７：抗CD206抗体（No.10）のHCDR1のアミノ酸配列
配列番号２８：抗CD206抗体（No.10）のHCDR2のアミノ酸配列
配列番号２９：抗CD206抗体（No.10）のHCDR3のアミノ酸配列
配列番号３０：抗CD206抗体（No.10）のLCDR1のアミノ酸配列
配列番号３１：抗CD206抗体（No.10）のLCDR3のアミノ酸配列
配列番号３２：抗CD206抗体（No.11）のHCDR1のアミノ酸配列
配列番号３３：抗CD206抗体（No.11）のHCDR2のアミノ酸配列
配列番号３４：抗CD206抗体（No.11）のHCDR3のアミノ酸配列
配列番号３５：抗CD206抗体（No.11）のLCDR1のアミノ酸配列
配列番号３６：抗CD206抗体（No.11）のLCDR3のアミノ酸配列
配列番号３７：抗CD206抗体（No.27）のHCDR1のアミノ酸配列
配列番号３８：抗CD206抗体（No.27）のHCDR2のアミノ酸配列
配列番号３９：抗CD206抗体（No.27）のHCDR3のアミノ酸配列
配列番号４０：抗CD206抗体（No.27）のLCDR1のアミノ酸配列
配列番号４１：抗CD206抗体（No.27）のLCDR3のアミノ酸配列
配列番号４２：抗CD206抗体（No.35）のHCDR1のアミノ酸配列
配列番号４３：抗CD206抗体（No.35）のHCDR2のアミノ酸配列
配列番号４４：抗CD206抗体（No.35）のHCDR3のアミノ酸配列
配列番号４５：抗CD206抗体（No.35）のLCDR1のアミノ酸配列
配列番号４６：抗CD206抗体（No.35）のLCDR3のアミノ酸配列
配列番号４７：抗CD206抗体（No.40）のHCDR1のアミノ酸配列
配列番号４８：抗CD206抗体（No.40）のHCDR2のアミノ酸配列
配列番号４９：抗CD206抗体（No.40）のHCDR3のアミノ酸配列
配列番号５０：抗CD206抗体（No.40）のLCDR1のアミノ酸配列
配列番号５１：抗CD206抗体（No.40）のLCDR3のアミノ酸配列
配列番号５２：抗CD206抗体（No.44）のHCDR1のアミノ酸配列
配列番号５３：抗CD206抗体（No.44）のHCDR2のアミノ酸配列
配列番号５４：抗CD206抗体（No.44）のHCDR3のアミノ酸配列
配列番号５５：抗CD206抗体（No.44）のLCDR1のアミノ酸配列
配列番号５６：抗CD206抗体（No.44）のLCDR3のアミノ酸配列
配列番号５７：抗CD206抗体（No.47）のHCDR1のアミノ酸配列
配列番号５８：抗CD206抗体（No.47）のHCDR2のアミノ酸配列
配列番号５９：抗CD206抗体（No.47）のHCDR3のアミノ酸配列
配列番号６０：抗CD206抗体（No.47）のLCDR1のアミノ酸配列
配列番号６１：抗CD206抗体（No.47）のLCDR3のアミノ酸配列
配列番号６２：抗CD206抗体（No.54）のHCDR1のアミノ酸配列
配列番号６３：抗CD206抗体（No.54）のHCDR2のアミノ酸配列
配列番号６４：抗CD206抗体（No.54）のHCDR3のアミノ酸配列
配列番号６５：抗CD206抗体（No.54）のLCDR1のアミノ酸配列
配列番号６６：抗CD206抗体（No.54）のLCDR3のアミノ酸配列
配列番号６７：抗CD206抗体（No.59）のHCDR1のアミノ酸配列
配列番号６８：抗CD206抗体（No.59）のHCDR2のアミノ酸配列
配列番号６９：抗CD206抗体（No.59）のHCDR3のアミノ酸配列
配列番号７０：抗CD206抗体（No.59）のLCDR1のアミノ酸配列
配列番号７１：抗CD206抗体（No.59）のLCDR3のアミノ酸配列
配列番号７２：抗CD206抗体（No.64）のHCDR1のアミノ酸配列
配列番号７３：抗CD206抗体（No.64）のHCDR2のアミノ酸配列
配列番号７４：抗CD206抗体（No.64）のHCDR3のアミノ酸配列
配列番号７５：抗CD206抗体（No.64）のLCDR1のアミノ酸配列
配列番号７６：抗CD206抗体（No.64）のLCDR3のアミノ酸配列
配列番号７７：抗CD206抗体（No.72）のHCDR1のアミノ酸配列
配列番号７８：抗CD206抗体（No.72）のHCDR2のアミノ酸配列
配列番号７９：抗CD206抗体（No.72）のHCDR3のアミノ酸配列
配列番号８０：抗CD206抗体（No.72）のLCDR1のアミノ酸配列
配列番号８１：抗CD206抗体（No.72）のLCDR3のアミノ酸配列
配列番号８２：抗CD206抗体（No.73）のHCDR1のアミノ酸配列
配列番号８３：抗CD206抗体（No.73）のHCDR2のアミノ酸配列
配列番号８４：抗CD206抗体（No.73）のHCDR3のアミノ酸配列
配列番号８５：抗CD206抗体（No.73）のLCDR1のアミノ酸配列
配列番号８６：抗CD206抗体（No.73）のLCDR3のアミノ酸配列
配列番号８７：抗CD206抗体（No.83）のHCDR1のアミノ酸配列
配列番号８８：抗CD206抗体（No.83）のHCDR2のアミノ酸配列
配列番号８９：抗CD206抗体（No.83）のHCDR3のアミノ酸配列
配列番号９０：抗CD206抗体（No.83）のLCDR1のアミノ酸配列
配列番号９１：抗CD206抗体（No.83）のLCDR3のアミノ酸配列
配列番９２：抗CD206抗体（No.10）の重鎖可変領域をコードする塩基配列
配列番号９３：抗CD206抗体（No.10）の軽鎖可変領域をコードする塩基配列
配列番号９４：抗CD206抗体（No.11）の重鎖可変領域をコードする塩基配列
配列番号９５：抗CD206抗体（No.11）の軽鎖可変領域をコードする塩基配列
配列番号９６：抗CD206抗体（No.27）の重鎖可変領域をコードする塩基配列
配列番号９７：抗CD206抗体（No.27）の軽鎖可変領域をコードする塩基配列
配列番号９８：抗CD206抗体（No.35）の重鎖可変領域をコードする塩基配列
配列番号９９：抗CD206抗体（No.35）の軽鎖可変領域をコードする塩基配列
配列番号１００：抗CD206抗体（No.40）の重鎖可変領域をコードする塩基配列
配列番号１０１：抗CD206抗体（No.40）の軽鎖可変領域をコードする塩基配列
配列番号１０２：抗CD206抗体（No.44）の重鎖可変領域をコードする塩基配列
配列番号１０３：抗CD206抗体（No.44）の軽鎖可変領域をコードする塩基配列
配列番号１０４：抗CD206抗体（No.47）の重鎖可変領域をコードする塩基配列
配列番号１０５：抗CD206抗体（No.47）の軽鎖可変領域をコードする塩基配列
配列番号１０６：抗CD206抗体（No.54）の重鎖可変領域をコードする塩基配列
配列番号１０７：抗CD206抗体（No.54）の軽鎖可変領域をコードする塩基配列
配列番号１０８：抗CD206抗体（No.59）の重鎖可変領域をコードする塩基配列
配列番号１０９：抗CD206抗体（No.59）の軽鎖可変領域をコードする塩基配列
配列番号１１０：抗CD206抗体（No.64）の重鎖可変領域をコードする塩基配列
配列番号１１１：抗CD206抗体（No.64）の軽鎖可変領域をコードする塩基配列
配列番号１１２：抗CD206抗体（No.72）の重鎖可変領域をコードする塩基配列
配列番号１１３：抗CD206抗体（No.72）の軽鎖可変領域をコードする塩基配列
配列番号１１４：抗CD206抗体（No.73）の重鎖可変領域をコードする塩基配列
配列番号１１５：抗CD206抗体（No.73）の軽鎖可変領域をコードする塩基配列
配列番号１１６：抗CD206抗体（No.83）の重鎖可変領域をコードする塩基配列
配列番号１１７：抗CD206抗体（No.83）の軽鎖可変領域をコードする塩基配列
配列番号１１８：マウスCD206コーディング領域の塩基配列 <Description of Sequence Listing>
SEQ ID NO: 1: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 2: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 3: Anti-CD206 antibody (No. 11) heavy chain variable region amino acid sequence SEQ ID NO: 4: light chain variable region amino acid sequence of anti-CD206 antibody (No. 11) SEQ ID NO: 5: heavy chain variable region amino acid sequence of anti-CD206 antibody (No. 27) SEQ ID NO: 6: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 27) SEQ ID NO: 7: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 35) SEQ ID NO: 8: Anti-CD206 antibody (No. 35) SEQ ID NO:9: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No.40) SEQ ID NO:10: Amino acid sequence of light chain variable region of anti-CD206 antibody (No.40) SEQ ID NO: 11: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 12: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 13: Anti-CD206 antibody (No. 47) SEQ ID NO: 14: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 15: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 16: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 17: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 59) SEQ ID NO: 18: Anti-CD206 antibody (No. 59) SEQ ID NO: 19: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 20: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 21: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 72) SEQ ID NO: 22: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 72) SEQ ID NO: 23: Anti-CD206 antibody (No. 73) SEQ ID NO: 24: Amino acid sequence of light chain variable region of anti-CD206 antibody (No. 73) SEQ ID NO: 25: Amino acid sequence of heavy chain variable region of anti-CD206 antibody (No. 83) SEQ ID NO: 26: Amino acid sequence of light chain variable region of anti-CD206 antibody (No.83) SEQ ID NO: 27: Amino acid sequence of HCDR1 of anti-CD206 antibody (No. 10) SEQ ID NO: 28: Anti-CD2 Amino acid sequence of HCDR2 of 06 antibody (No.10) SEQ ID NO:29: Amino acid sequence of HCDR3 of anti-CD206 antibody (No.10) SEQ ID NO:30: Amino acid sequence of LCDR1 of anti-CD206 antibody (No.10) SEQ ID NO:31: LCDR3 amino acid sequence of anti-CD206 antibody (No.10) SEQ ID NO:32: HCDR1 amino acid sequence of anti-CD206 antibody (No.11) SEQ ID NO:33: HCDR2 amino acid sequence of anti-CD206 antibody (No.11) SEQ ID NO:34 : amino acid sequence of HCDR3 of anti-CD206 antibody (No.11) SEQ ID NO:35: amino acid sequence of LCDR1 of anti-CD206 antibody (No.11) SEQ ID NO:36: amino acid sequence of LCDR3 of anti-CD206 antibody (No.11) SEQ ID NO: 37: HCDR1 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO:38: HCDR2 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO:39: HCDR3 amino acid sequence of anti-CD206 antibody (No.27) No. 40: LCDR1 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO: 41: LCDR3 amino acid sequence of anti-CD206 antibody (No.27) SEQ ID NO: 42: HCDR1 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 43: HCDR2 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 44: HCDR3 amino acid sequence of anti-CD206 antibody (No.35) SEQ ID NO: 45: LCDR1 amino acid of anti-CD206 antibody (No.35) Sequences SEQ ID NO: 46: LCDR3 amino acid sequence of anti-CD206 antibody (No. 35) SEQ ID NO: 47: HCDR1 amino acid sequence of anti-CD206 antibody (No. 40) SEQ ID NO: 48: HCDR2 of anti-CD206 antibody (No. 40) Amino acid sequences SEQ ID NO: 49: Amino acid sequence of HCDR3 of anti-CD206 antibody (No. 40) SEQ ID NO: 50: Amino acid sequence of LCDR1 of anti-CD206 antibody (No. 40) SEQ ID NO: 51: LCDR3 of anti-CD206 antibody (No. 40) SEQ ID NO: 52: HCDR1 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 53: HCDR2 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 54: anti-CD206 antibody (No.44) HCDR3 amino acid sequence SEQ ID NO: 55: LCDR1 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: No. 56: LCDR3 amino acid sequence of anti-CD206 antibody (No.44) SEQ ID NO: 57: HCDR1 amino acid sequence of anti-CD206 antibody (No.47) SEQ ID NO: 58: HCDR2 amino acid sequence of anti-CD206 antibody (No.47) SEQ ID NO: 59: HCDR3 amino acid sequence of anti-CD206 antibody (No. 47) SEQ ID NO: 60: LCDR1 amino acid sequence of anti-CD206 antibody (No. 47) SEQ ID NO: 61: LCDR3 amino acid of anti-CD206 antibody (No. 47) Sequences SEQ ID NO:62: HCDR1 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO:63: HCDR2 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO:64: HCDR3 of anti-CD206 antibody (No.54) Amino acid sequences SEQ ID NO: 65: LCDR1 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO: 66: LCDR3 amino acid sequence of anti-CD206 antibody (No.54) SEQ ID NO: 67: HCDR1 of anti-CD206 antibody (No.59) SEQ ID NO: 68: HCDR2 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO: 69: HCDR3 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO: 70: anti-CD206 antibody (No.59) LCDR1 amino acid sequence SEQ ID NO:71: LCDR3 amino acid sequence of anti-CD206 antibody (No.59) SEQ ID NO:72: HCDR1 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:73: anti-CD206 antibody (No.64) SEQ ID NO:74: HCDR3 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:75: LCDR1 amino acid sequence of anti-CD206 antibody (No.64) SEQ ID NO:76: anti-CD206 antibody (No.64 ) SEQ ID NO: 77: HCDR1 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 78: HCDR2 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 79: anti-CD206 antibody (No. 72) HCDR3 amino acid sequence SEQ ID NO: 80: LCDR1 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 81: LCDR3 amino acid sequence of anti-CD206 antibody (No. 72) SEQ ID NO: 82: anti-CD206 antibody (No. 73) HCDR1 amino acid sequence SEQ ID NO:83: HCDR2 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 84: HCDR3 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 85: LCDR1 amino acid sequence of anti-CD206 antibody (No.73) SEQ ID NO: 86: anti-CD206 antibody (No.73) LCDR3 amino acid sequence SEQ ID NO:87: HCDR1 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:88: HCDR2 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:89: anti-CD206 antibody (No.83) SEQ ID NO:90: LCDR1 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:91: LCDR3 amino acid sequence of anti-CD206 antibody (No.83) SEQ ID NO:92: anti-CD206 antibody (No.10 ) SEQ ID NO: 93: Nucleotide sequence encoding the light chain variable region of anti-CD206 antibody (No. 10) SEQ ID NO: 94: Heavy chain variable region of anti-CD206 antibody (No. 11) SEQ ID NO:95: Nucleotide sequence encoding the light chain variable region of anti-CD206 antibody (No.11) SEQ ID NO:96: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.27) No. 97: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 27) SEQ ID NO: 98: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 35) SEQ ID NO: 99: Anti-CD206 antibody Nucleotide sequence encoding the light chain variable region of (No.35) SEQ ID NO: 100: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No.40) SEQ ID NO: 101: Anti-CD206 antibody (No.40) Nucleotide sequence encoding light chain variable region SEQ ID NO: 102: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 103: Encoding light chain variable region of anti-CD206 antibody (No. 44) SEQ ID NO: 104: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 105: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 47) SEQ ID NO: 106 : Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.54) SEQ ID NO: 107: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 54) SEQ ID NO: 108: Anti-CD206 antibody (No. 59) SEQ ID NO: 109: anti-CD206 antibody (No 59) SEQ ID NO: 110: Nucleotide sequence encoding the heavy chain variable region of anti-CD206 antibody (No. 64) SEQ ID NO: 111: Light chain of anti-CD206 antibody (No. 64) Nucleotide sequence encoding variable region SEQ ID NO: 112: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.72) SEQ ID NO: 113: Nucleotide encoding light chain variable region of anti-CD206 antibody (No.72) Sequence SEQ ID NO: 114: Nucleotide sequence encoding heavy chain variable region of anti-CD206 antibody (No.73) SEQ ID NO: 115: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No.73) SEQ ID NO: 116: Anti Nucleotide sequence encoding heavy chain variable region of CD206 antibody (No.83) SEQ ID NO: 117: Nucleotide sequence encoding light chain variable region of anti-CD206 antibody (No. 83) SEQ ID NO: 118: Nucleotide sequence of mouse CD206 coding region
Claims (8)
- CD206に特異的に結合する抗体またはその機能的断片であって、
前記抗体が以下から選択される抗体である、抗体またはその機能的断片：
（ａ）配列番号２７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号２８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号２９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号３０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号３１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｂ）配列番号３７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号３８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号３９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＬＭＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｃ）配列番号４２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号４５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号４６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｄ）配列番号４７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号４８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号４９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＦＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｅ）配列番号５２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号５５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＡＴで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号５６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｆ）配列番号５７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号５８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号５９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｇ）配列番号６２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号６５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＫＡＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号６６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｈ）配列番号６７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号６８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号６９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号７０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号７１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｉ）配列番号７７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号７８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号７９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＡＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；
（ｊ）配列番号８２で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８３で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８４で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号８５で表されるアミノ酸配列からなる軽鎖LCDR1、
ＲＡＮで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号８６で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体；並びに
（ｋ）配列番号８７で表されるアミノ酸配列からなる重鎖HCDR1、
配列番号８８で表されるアミノ酸配列からなる重鎖HCDR2、
配列番号８９で表されるアミノ酸配列からなる重鎖HCDR3、
配列番号９０で表されるアミノ酸配列からなる軽鎖LCDR1、
ＹＴＳで表されるアミノ酸配列からなる軽鎖LCDR2、および
配列番号９１で表されるアミノ酸配列からなる軽鎖LCDR3を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206,
An antibody or functional fragment thereof, wherein said antibody is an antibody selected from:
(a) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 27;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 28;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 29;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 30;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:31;
(b) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 37;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 38;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 39;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 40;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by LMS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 41;
(c) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 42;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 43;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 44;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 45;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 46;
(d) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 47;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 48;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 49;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 50;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by FAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:51;
(e) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 52;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 53;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 54;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 55;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by AAT and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:56;
(f) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 57;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 58;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 59;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 60;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 61;
(g) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 62;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 63;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 64;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 65;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by KAS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:66;
(h) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 67;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 68;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 69;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 70;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:71;
(i) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 77;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 78;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 79;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 80;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by ATS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:81;
(j) heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO:82;
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 83;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 84;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 85;
an antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by RAN and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 86; and (k) a heavy chain HCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 87 ,
heavy chain HCDR2 consisting of the amino acid sequence represented by SEQ ID NO: 88;
heavy chain HCDR3 consisting of the amino acid sequence represented by SEQ ID NO: 89;
a light chain LCDR1 consisting of the amino acid sequence represented by SEQ ID NO: 90;
An antibody comprising a light chain LCDR2 consisting of the amino acid sequence represented by YTS and a light chain LCDR3 consisting of the amino acid sequence represented by SEQ ID NO:91. - 前記抗体が以下から選択される抗体である、請求項１に記載のCD206に特異的に結合する抗体またはその機能的断片：
（Ａ）配列番号１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｂ）配列番号５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｃ）配列番号７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｄ）配列番号９で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１０で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｅ）配列番号１１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｆ）配列番号１３で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１４で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｇ）配列番号１５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｈ）配列番号１７で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号１８で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｉ）配列番号２１で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２２で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；
（Ｊ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体；並びに
（Ｋ）配列番号２５で表されるアミノ酸配列からなる重鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する重鎖可変領域、および
配列番号２６で表されるアミノ酸配列からなる軽鎖可変領域または該アミノ酸配列と90％以上の配列同一性を有するアミノ酸配列からなり、CD206への結合活性を有する軽鎖可変領域を含む抗体。 An antibody or functional fragment thereof that specifically binds to CD206 according to claim 1, wherein said antibody is an antibody selected from:
(A) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 1 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 2 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(B) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 5 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 6 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(C) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 7 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(D) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(E) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 11 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(F) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 13 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 14 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(G) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 15 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 16 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(H) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 17 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 18 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(I) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 21 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity;
(J) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; an antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; (K) a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 25 or a heavy chain variable region consisting of an amino acid sequence having a sequence identity of 90% or more with the amino acid sequence and having CD206-binding activity; An antibody comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 26 or a light chain variable region consisting of an amino acid sequence having 90% or more sequence identity with said amino acid sequence and having CD206-binding activity. - 請求項１または２に記載のCD206に特異的に結合する抗体またはその機能的断片をコードする、単離されたDNA。 An isolated DNA encoding the antibody that specifically binds to CD206 according to claim 1 or 2 or a functional fragment thereof.
- 請求項１または２に記載のCD206に特異的に結合する抗体またはその機能的断片を有効成分として含む、CD206陽性M2マクロファージ細胞殺傷剤。 A CD206-positive M2 macrophage cell-killing agent comprising, as an active ingredient, the antibody that specifically binds to CD206 according to claim 1 or 2 or a functional fragment thereof.
- CDC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、請求項４に記載のCD206陽性M2マクロファージ細胞殺傷剤。 The CD206-positive M2 macrophage cell-killing agent according to claim 4, which has CD206-positive M2 macrophage cytotoxicity due to CDC activity.
- ADCC活性によるCD206陽性M2マクロファージ細胞障害作用を有する、請求項４に記載のCD206陽性M2マクロファージ細胞殺傷剤。 The CD206-positive M2 macrophage cell-killing agent according to claim 4, which has CD206-positive M2 macrophage cytotoxicity due to ADCC activity.
- 請求項１または２に記載のCD206に特異的に結合する抗体またはその機能的断片を有効成分として含む、代謝障害の予防または治療のための医薬組成物。 A pharmaceutical composition for preventing or treating metabolic disorders, comprising the antibody that specifically binds to CD206 according to claim 1 or 2 or a functional fragment thereof as an active ingredient.
- 前記代謝障害が、耐糖機能障害、インスリン抵抗性および糖尿病から選択される１種以上である、請求項７に記載の医薬組成物。 The pharmaceutical composition according to claim 7, wherein the metabolic disorder is one or more selected from impaired glucose tolerance, insulin resistance and diabetes.
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