WO2018235247A1 - Anticorps anti-cd10 - Google Patents

Anticorps anti-cd10 Download PDF

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WO2018235247A1
WO2018235247A1 PCT/JP2017/023104 JP2017023104W WO2018235247A1 WO 2018235247 A1 WO2018235247 A1 WO 2018235247A1 JP 2017023104 W JP2017023104 W JP 2017023104W WO 2018235247 A1 WO2018235247 A1 WO 2018235247A1
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antibody
present
amino acid
cells
acid sequence
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PCT/JP2017/023104
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Japanese (ja)
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奈津子 水谷
雅明 阿部
周二 松岡
興夫 樋野
奈穂美 大辻
良 波多野
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学校法人順天堂
日本抗体医薬株式会社
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Priority to JP2019524814A priority Critical patent/JP7002145B2/ja
Priority to PCT/JP2017/023104 priority patent/WO2018235247A1/fr
Publication of WO2018235247A1 publication Critical patent/WO2018235247A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material
    • C12N5/12—Fused cells, e.g. hybridomas

Definitions

  • the present invention relates to anti-CD10 antibodies.
  • CD10 is a membrane protein expressed on the surface of cells. Clinically, it is used, for example, as a tumor marker for B cell leukemia.
  • Non-Patent Document 1 describes that CD10 was expressed in leukemic blasts of a patient with pediatric acute lymphoblastic leukemia.
  • NL-1 is used as an antibody for detecting CD10.
  • Non-Patent Document 2 describes that NCL-CD10-270 was produced as a new antibody that recognizes CD10 of paraffin-embedded tissue.
  • CD10 has many unexplained parts, and some anti-CD10 antibodies have been reported as only some of the antibodies described in the above Non-patent Documents 1 and 2. In particular, with regard to anti-CD10 antibodies for the treatment of malignant tumors, sufficient research results have not been obtained.
  • the present invention has been made in view of the above circumstances, and it is an object of the present invention to provide an anti-CD10 antibody that exhibits excellent effects in the treatment of malignant tumors.
  • the present inventors conducted experiments to administer NEP1 antibody, which is an anti-CD10 antibody, to tumor-bearing mice, as described in the examples below. As a result, the survival time of tumor-bearing mice was surprisingly extended. And based on this result, the present invention was completed.
  • an anti-CD10 antibody having an amino acid sequence identical to the amino acid sequences of heavy chain CDRs 1 to 3 and light chain CDRs 1 to 3 of the antibody produced by the hybridoma of Accession No. NITE BP-02489 Provided.
  • This antibody can be used to treat malignancy.
  • a pharmaceutical composition for treating a malignant tumor which comprises the above-described anti-CD10 antibody.
  • This pharmaceutical composition can be used to treat malignant tumors.
  • a polynucleotide or a vector encoding the above-described anti-CD10 antibody a polynucleotide or a vector encoding the above-described anti-CD10 antibody.
  • the above-described anti-CD10 antibody can be produced using this polynucleotide or vector.
  • a cell comprising a polynucleotide encoding the above anti-CD10 antibody.
  • the cells can be used to produce the anti-CD10 antibody.
  • FIG. 1 shows the results of Western blot.
  • FIG. 2 shows the results of FACS analysis.
  • FIG. 3 shows the results of a tumor-bearing mouse treatment experiment.
  • the anti-CD10 antibody according to one embodiment of the present invention is an anti-CD10 antibody, for example, an anti-CD10 antibody having an amino acid sequence identical to the amino acid sequences of heavy chain CDRs 1-3 and light chain CDRs 1-3 of the antibody produced by NITE BP-02489 hybridoma.
  • CD10 antibody This anti-CD10 antibody can prolong the survival time of tumor-bearing mice, as demonstrated in the examples below. Since this anti-CD10 antibody shows therapeutic effect in animal experiments, it has a large clinical contribution in the treatment of malignant tumors. Moreover, this anti-CD10 antibody has an unexpectedly high therapeutic effect on malignant tumors.
  • CD10 is known as a membrane protein expressed on the surface of cells.
  • the CD 10 also includes other names such as CALLA, Neprilysin, etc., as long as the structure is substantially the same.
  • the amino acid sequence of CD10 may be, for example, the amino acid sequence of a protein represented by NCBI Accession No. NP — 009220.2 or UniProt KB Accession No. P08473.
  • an "anti-CD10 antibody” comprises an antibody having binding to CD10.
  • the method of producing this anti-CD10 antibody is not particularly limited, for example, it may be produced using a hybridoma of Accession No. NITE BP-02489.
  • the analysis of the base sequence of the nucleic acid encoding the variable region (including CDRs) of the anti-CD10 antibody produced from the hybridoma of accession number NITE BP-02489 can be performed, for example, through the following steps.
  • an "antibody” comprises a molecule or population thereof capable of specifically binding to a particular epitope on an antigen.
  • the antibody can exist in various forms, for example, full-length antibody (antibody having Fab region and Fc region), Fv antibody, Fab antibody, F (ab ') 2 antibody, Fab' antibody, diabody, single antibody Chain antibody (eg, scFv), dsFv, multivalent specific antibody (eg, bispecific antibody), peptide or polypeptide with antigen binding, chimeric antibody, mouse antibody, humanized antibody, human antibody, or those It may be one or more forms selected from the group consisting of the equivalent (or equivalent) of Antibodies also include modified or unmodified antibodies.
  • the modified antibody may be bound to an antibody and various molecules such as polyethylene glycol.
  • Modified antibodies can be obtained by chemically modifying antibodies using known techniques.
  • the antibody may also be a fusion protein.
  • the fusion protein may be one in which a polypeptide or an oligopeptide (eg, His tag) is linked to the N or C terminus of the antibody.
  • the fusion protein may also be a fusion of antibody partial sequences such as mouse or human.
  • the antibody may also be a drug-bound antibody.
  • the drug may be, for example, a cytotoxic agent or an anti-cancer agent.
  • Such modified antibodies, fusion proteins, antibody drug complexes are also included in one form of antibody.
  • the amino acid sequence, class or subclass of the antibody may be derived from, for example, humans or mammals other than humans (eg, rats, mice, rabbits, cattle, monkeys, etc.).
  • the antibody class is not particularly limited, and may be, for example, IgG, IgA, IgM, IgD or IgE.
  • the antibody subclass is not particularly limited, and may be, for example, IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2.
  • Antibodies also include, for example, isolated antibodies, purified antibodies, or recombinant antibodies. The antibodies can also be used, for example, in vitro or in vivo.
  • the "polyclonal antibody” can be generated, for example, by administering an immunogen containing an antigen of interest to mammals (eg, rats, mice, rabbits, cattle, monkeys, etc.) and the like. It is.
  • the administration of the immunogen may be co-infused with one or more immunizing agents, or an adjuvant.
  • Immunization protocols are known in the art, and are carried out by any method that induces an immune response, depending on the host organism etc. to be selected (Protein Experimental Handbook, Yodosha (2003): 86-91.).
  • monoclonal antibodies include antibodies wherein individual antibodies making up a population respond to substantially the same epitope.
  • the individual antibodies that make up the population may be antibodies that are substantially identical (naturally occurring mutations are acceptable).
  • the method for producing a monoclonal antibody is not particularly limited, but, for example, a method similar to the hybridoma method as disclosed in "Kohler G, Milstein C., Nature. 1975 Aug 7; 256 (5517): 495-497.” It may be produced by Alternatively, monoclonal antibodies may be made by methods similar to recombinant methods as described in US Pat. No. 4,816,567. Alternatively, the monoclonal antibody may be “Clackson et al., Nature.
  • an "Fv antibody” is an antibody that comprises an antigen recognition site. This region contains a dimer of one heavy chain variable domain and one light chain variable domain by noncovalent binding. In this configuration, the three CDRs of each variable domain can interact to form an antigen binding site on the surface of the VH-VL dimer.
  • the “Fab antibody” is, for example, about N-terminal half of the H chain and the entire L chain among fragments obtained by treating an antibody containing the Fab region and the Fc region with the proteolytic enzyme papain Is an antibody linked through some disulfide bonds.
  • Fab can be obtained, for example, by treating the anti-CD10 antibody according to the above-mentioned embodiment of the present invention containing a Fab region and an Fc region with the proteolytic enzyme papain.
  • F (ab ′) 2 antibody is, for example, a fragment corresponding to Fab in a fragment obtained by treating an antibody containing Fab region and Fc region with proteolytic enzyme pepsin
  • Antibodies that contain F (ab ′) 2 can be obtained, for example, by treating the anti-CD10 antibody according to the above-mentioned embodiment of the present invention containing a Fab region and an Fc region with the proteolytic enzyme pepsin.
  • it can be produced by thioether bond or disulfide bond of the following Fab ′.
  • the “Fab ′ antibody” is, for example, an antibody obtained by cleaving a disulfide bond in the hinge region of F (ab ′) 2 .
  • F (ab ') 2 can be obtained by treatment with a reducing agent dithiothreitol.
  • the "scFv antibody” is an antibody in which VH and VL are linked via a suitable peptide linker.
  • the scFv antibody obtains cDNAs encoding VH and VL of the anti-CD10 antibody according to the above-mentioned embodiment of the present invention, constructs a polynucleotide encoding VH-peptide linker-VL, and prepares the polynucleotide as a vector And can be produced using cells for expression.
  • a "diabody” is an antibody with bivalent antigen binding activity.
  • the bivalent antigen binding activities can be identical or one can be different antigen binding activities.
  • a polynucleotide encoding a scFv is constructed so that the amino acid sequence of the peptide linker has a length of 8 residues or less, and the obtained polynucleotide is incorporated into a vector and produced using cells for expression it can.
  • dsFv is an antibody in which a polypeptide having a cysteine residue introduced into VH and VL is linked via a disulfide bond between the above-mentioned cysteine residues.
  • the position to be introduced into the cysteine residue should be selected based on the prediction of the three-dimensional structure of the antibody according to the method shown by Reiter et al. (Reiter et al., Protein Eng. 1994 May; 7 (5): 697-704.). Can.
  • the “peptide or polypeptide having antigen binding property” is an antibody comprising VH, VL of the antibody, or CDRs 1, 2 or 3 thereof. Peptides containing multiple CDRs can be linked directly or via appropriate peptide linkers.
  • Fv antibody The above-mentioned Fv antibody, Fab antibody, F (ab ') 2 antibody, Fab' antibody, scFv antibody, diabody, dsFv antibody, peptide or polypeptide having antigen binding ability (hereinafter referred to as "Fv antibody etc.”
  • the production method of) is not particularly limited.
  • a DNA encoding a region such as an Fv antibody in the anti-CD10 antibody according to the embodiment of the present invention described above can be incorporated into an expression vector and produced using expression cells. Alternatively, they may be produced by a chemical synthesis method such as Fmoc method (fluorenylmethyloxycarbonyl method), tBOC method (t-butyloxycarbonyl method) or the like.
  • a "chimeric antibody” is, for example, one obtained by linking the variable region of an antibody between heterologous organisms and a constant region, and can be constructed by genetic recombination technology.
  • a mouse-human chimeric antibody can be produced, for example, by the method described in "Roguska et al., Proc Natl Acad Sci US A. 1994 Feb 1; 91 (3) 969-973.”.
  • a basic method for producing mouse-human chimeric antibodies is, for example, encoding the mouse leader and variable region sequences present in the cloned cDNA, and human antibody constant regions already present in mammalian cell expression vectors.
  • the mouse leader sequence and variable region sequence present in the cloned cDNA may be ligated to a sequence encoding a human antibody constant region and then ligated to a mammalian cell expression vector.
  • the fragments of the human antibody constant region can be those of the heavy chain constant region of any human antibody and the light chain constant region of the human antibody, eg, for human heavy chain, C ⁇ 1, C ⁇ 2, C ⁇ 3 or C ⁇ 4 As for the L chain one, C ⁇ or C ⁇ ⁇ ⁇ can be mentioned respectively.
  • a "humanized antibody” has, for example, one or more CDRs from a non-human species, and a framework region derived from human immunoglobulin, and further a constant region derived from human immunoglobulin.
  • CDR grafting (Ozaki et al., Blood. 1999 Jun 1; 93 (11): 3922-3930.), Re-surfacing (roguska et al., Proc Natl Acad Sci USA A.
  • FR shuffle (Damschroder et al., Mol Immunol. 2007 Apr; 44 (11): 3049-3060. Epub 2007 Jan 22.) and the like.
  • Amino acid residues of the human FR region may be substituted with the corresponding residues from the CDR donor antibody to alter or improve antigen binding. This FR substitution can be carried out by methods well known in the art (Riechmann et al., Nature. 1988 Mar 24; 332 (6162): 323-327.).
  • FR residues important for antigen binding may be identified by modeling the interaction of CDRs with FR residues.
  • abnormal FR residues may be identified at specific positions by sequence comparison.
  • variable region In one embodiment of the invention "heavy chain” is typically the main component of full-length antibodies.
  • the heavy chain is usually linked to the light chain by disulfide bonds and noncovalent bonds.
  • VH variable region
  • the amino acid sequence is not constant even in the same kind of antibody of the same class.
  • VH has a large affinity to the antigen. It is known to contribute.
  • a molecule consisting only of VH was produced in "Reiter et al., J Mol Biol. 1999 Jul 16; 290 (3): 685-98.”, It specifically bound to the antigen with high affinity. Is described.
  • Wolfson W Chem Biol. 2006 Dec; 13 (12): 1243-1244.
  • CDR complementarity determining region
  • Fv variable region: including heavy chain variable region (VH) and light chain variable region (VL)
  • CDRs include CDR1, CDR2 and CDR3 consisting of about 5 to 30 amino acid residues.
  • heavy chain CDRs particularly contribute to antibody binding to antigen.
  • CDR3 is known to have the highest contribution in antibody binding to antigen.
  • CDR CDR
  • the definition of CDR and the method for determining the position thereof are described in Kabat (Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. (1991)), or Chothia.
  • the definition (Chothia et al., J. Mol. Biol., 1987; 196: 901-917) may be adopted.
  • the definition of Kabat is taken as a preferred example.
  • it may be determined in consideration of both the definition of Kabat and the definition of Chothia for example, a portion including overlapping portions of CDRs according to each definition or portions including both CDRs according to each definition It can also be a CDR.
  • amino acid is a generic term for organic compounds having an amino group and a carboxyl group.
  • any amino acid in the amino acid sequence may be chemically modified.
  • any amino acid in the amino acid sequence may form a salt or a solvate.
  • any amino acid in the amino acid sequence may be L-form or D-form. Even in those cases, it can be said that the antibody according to the embodiment of the present invention comprises the above-mentioned "specific amino acid sequence".
  • Examples of chemical modifications that amino acids contained in proteins undergo in vivo include, for example, N-terminal modification (eg, acetylation, myristoylation etc.), C-terminal modification (eg amidification, glycosylphosphatidylinositol addition etc.), or side chain Modifications (eg, phosphorylation, glycosylation, etc.) and the like are known.
  • N-terminal modification eg, acetylation, myristoylation etc.
  • C-terminal modification eg amidification, glycosylphosphatidylinositol addition etc.
  • side chain Modifications eg, phosphorylation, glycosylation, etc.
  • One embodiment of the present invention is a polynucleotide or a vector encoding an anti-CD10 antibody according to the above-mentioned embodiment of the present invention.
  • Transformants can be produced by introducing this polynucleotide or vector into cells.
  • the transformant may be a cell of human or a mammal other than human (eg, rat, mouse, guinea pig, rabbit, bovine, monkey, etc.).
  • Mammalian cells include, for example, Chinese hamster ovary cells (CHO cells), monkey cells COS-7, human fetal kidney cells (eg, HEK 293 cells) and the like.
  • the transformant may be Escherichia genus, yeast or the like.
  • the polynucleotide or vector may be constructed so as to express an anti-CD10 antibody.
  • the polynucleotide or vector may contain, for example, components necessary for protein expression, such as a promoter, an enhancer, an origin of replication, or an antibiotic resistance gene.
  • the vector examples include a plasmid derived from E. coli (eg, pET-Blue), a plasmid derived from Bacillus subtilis (eg, pUB110), a yeast-derived plasmid (eg, pSH19), an animal cell expression plasmid (eg, pA1-11, pcDNA3.1- Bacteriophages such as V5 / His-TOPO), ⁇ phage, and vectors derived from viruses can be used.
  • the vector may be an expression vector or may be circular.
  • a method for introducing the above polynucleotide or vector into cells for example, calcium phosphate method, lipofection method, electroporation method, method by adenovirus, method by retrovirus, microinjection, etc. can be used (revised 4 th ed. New Genetic Engineering Handbook, Yodosha (2003): 152-179.).
  • a method for producing an antibody using cells for example, the method described in "Protein Experimental Handbook, Yodosha (2003): 128-142.” Can be used.
  • One embodiment of the present invention is a method for producing an anti-CD10 antibody, which comprises the step of growing cells containing the polynucleotide or vector according to the above-mentioned embodiment of the present invention.
  • the growing step includes a culturing step.
  • the production method may also include the step of recovering the anti-CD10 antibody.
  • the production method may also include the step of preparing a cell culture solution.
  • the production method may also include the step of purifying the anti-CD10 antibody.
  • purification of the antibody is carried out, for example, with ammonium sulfate, ethanol precipitation, protein A, protein G, gel filtration chromatography, anion, cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography Chromatography, affinity chromatography, hydroxylapatite chromatography, or lectin chromatography can be used (Protein Experimental Handbook, Yodosha (2003): 27-52.).
  • One embodiment of the present invention is a composition comprising the anti-CD10 antibody according to the above-mentioned embodiment of the present invention.
  • This composition can be used to treat malignancy.
  • One or more of various embodiments for example, the possibility of containing a carrier, etc.
  • the pharmaceutical composition described later are applicable to this composition.
  • One embodiment of the present invention is a pharmaceutical composition comprising the anti-CD10 antibody according to the above-mentioned embodiment of the present invention.
  • This pharmaceutical composition can be used to treat malignant tumors.
  • the pharmaceutical composition may comprise one or more pharmacologically acceptable carriers.
  • the pharmaceutical composition includes, for example, a pharmaceutical composition for treating a malignant tumor.
  • One embodiment of the present invention is a method for treating a malignant tumor, which comprises the step of administering to a patient an anti-CD10 antibody (or a pharmaceutical composition comprising an anti-CD10 antibody) according to the above-mentioned embodiment of the present invention.
  • One embodiment of the present invention is the use of the anti-CD10 antibody according to the above-mentioned embodiment of the present invention for producing a pharmaceutical composition.
  • malignant tumor includes, for example, a tumor which is generated by mutation of a normal cell. Malignant tumors can arise from any organ or tissue throughout the body.
  • the malignancy includes, for example, mesothelioma or malignant lymphoma.
  • the mesothelioma includes mesothelial cell-derived tumors.
  • Mesothelioma also includes, for example, pleural mesothelioma, peritoneal mesothelioma, pericardial mesothelioma, testicular mesothelioma.
  • the malignant lymphoma includes, for example, Hodgkin's lymphoma and non-Hodgkin's lymphoma.
  • treatment includes the ability to exert a symptom ameliorating effect, a suppressing effect, a recurrence suppressing effect or a preventing effect of a disease of a patient or one or more symptoms associated with the disease.
  • the “pharmaceutical composition” is produced, for example, by mixing the active ingredient and one or more pharmacologically acceptable carriers, and producing it by any method known in the pharmaceutical arts. May be The use of the pharmaceutical composition is not limited as long as it is used for treatment, and the active ingredient may be used alone, or may be a mixture of the active ingredient and an optional ingredient.
  • the shape of the carrier is not particularly limited, and may be, for example, a solid or a liquid (for example, buffer).
  • the content of the carrier may be, for example, a pharmaceutically effective amount.
  • the effective amount may be, for example, an amount sufficient for pharmaceutical stability or delivery of the active ingredient.
  • buffers are effective in stabilizing the active ingredient in the vial.
  • the route of administration of the pharmaceutical composition is preferably that which is effective in treatment, and may be, for example, intravenous administration.
  • the administration mode may be, for example, an injection.
  • Aqueous solutions for injection may, for example, be stored in vials.
  • the aqueous solution for injection may also be formulated with, for example, physiological saline or a buffer (eg, phosphate buffer).
  • the dose, administration interval, and administration method are not particularly limited, but can be appropriately selected depending on the age and weight of the patient, symptoms, target organs and the like.
  • the pharmaceutical composition also preferably contains a therapeutically effective amount, or an effective amount of an active ingredient that exerts a desired effect.
  • the therapeutic effect of the pharmaceutical composition may be evaluated, for example, by prolonging the survival time of tumor-bearing mice. In this case, for example, a test is performed on 7 tumor-bearing mice, and all the PBS-administered groups die on day 35 after the administration start date, and 5 or more in the anti-CD10 antibody group are alive. You may judge that there is an effect.
  • the therapeutic effect may also be evaluated by diagnostic imaging.
  • the “patient” is a human or a non-human mammal (eg, mouse, guinea pig, hamster, rat, rat, rat, rabbit, pig, sheep, goat, cow, horse, cat, dog, marmoset, Monkeys or one or more species such as chimpanzees).
  • the patient may also be a patient diagnosed as having a malignancy or a patient in need of treatment.
  • the anti-CD10 antibody according to one embodiment of the present invention comprises a monoclonal antibody.
  • a monoclonal antibody it can be efficiently acted on CD10 as compared to a polyclonal antibody.
  • An anti-CD10 antibody according to an embodiment of the present invention comprises an antibody fragment having CD10 binding activity (hereinafter sometimes referred to as “antigen binding fragment”).
  • antigen binding fragment may be one or more of the above-mentioned Fv antibody and the like.
  • the anti-CD10 antibody according to one embodiment of the present invention has, for example, an amino acid sequence identical to the amino acid sequence of the heavy chain variable region and light chain variable region of the antibody produced by the hybridoma of Accession No. NITE BP-02489 May be
  • the anti-CD10 antibody can also be used, for example, heavy chain FR1, 2, 3, 4, light chain FR1, 2, 3, 4, heavy chain Fab, light chain Fab, of the antibody produced by the hybridoma of Accession No. NITE BP-02489. It may have the same amino acid sequence as that of the Fc region, heavy chain or light chain.
  • the anti-CD10 antibody according to one embodiment of the present invention comprises an anti-CD10 antibody that suppresses the growth of malignant tumor cells.
  • the NEP1 antibody described later has a function of suppressing the growth of malignant tumor cells.
  • the anti-CD10 antibody according to one embodiment of the present invention includes an anti-CD10 antibody that suppresses the growth of malignant mesothelioma cells or malignant lymphoma cells.
  • the NEP1 antibody described below has a function of suppressing the growth of malignant mesothelioma cells or malignant lymphoma cells.
  • Malignant tumor cells may express CD10.
  • the anti-CD10 antibody according to one embodiment of the present invention includes an anti-CD10 antibody that recognizes CD10 expressed in malignant tumor cells.
  • An anti-CD10 antibody according to one embodiment of the present invention also includes an antibody that binds to a wild type or a mutant form of CD10. Variants include those that result from differences in DNA sequences between individuals, such as SNPs.
  • One embodiment of the present invention is a pharmaceutical composition for treating a malignant tumor, which comprises an anti-CD10 antibody.
  • This pharmaceutical composition can be used to treat malignant tumors.
  • One embodiment of the present invention is a growth inhibitor of malignant tumor, which comprises an anti-CD10 antibody. This growth inhibitor can be used to treat malignancy.
  • One embodiment of the present invention is a diagnostic agent for malignant tumor, which comprises the anti-CD10 antibody according to the above-mentioned embodiment of the present invention. Malignant tumors can be diagnosed efficiently by using this diagnostic agent.
  • One embodiment of the present invention is a method for diagnosing a malignant tumor, comprising the step of contacting a patient sample with the anti-CD10 antibody according to the above-mentioned embodiment of the present invention.
  • One embodiment of the present invention is a detection reagent for CD10, which comprises the anti-CD10 antibody according to the above-mentioned embodiment of the present invention. By using this reagent, CD10 can be detected efficiently.
  • One embodiment of the present invention is a method of detecting CD10, comprising the step of contacting a test sample with the anti-CD10 antibody according to the above-mentioned embodiment of the present invention.
  • One embodiment of the present invention is a kit comprising the anti-CD10 antibody according to the above-mentioned embodiment of the present invention. With this kit, for example, treatment of diseases, diagnosis or detection of CD10 can be performed.
  • the kit may, for example, comprise the composition, the pharmaceutical composition, the diagnostic agent, or the detection reagent according to the embodiments of the present invention described above, and the instruction manual, the buffer solution, the container Or a package may be included.
  • CDR 1-3 means CDR1, CDR2, and CDR3.
  • “or” is used when “at least one or more” of the items listed in the text can be adopted. The same applies to “or”. In the present specification, when “within the range of two values” is specified, the range also includes the two values themselves. As used herein, “A to B” includes A and B.
  • Example 1 Analysis by Western Blotting HUH-7 cells (hepatocellular carcinoma, obtained from RIKEN Cell Bank (Tsukuba City, Ibaraki Prefecture)) and its CD10 transfectant (HuH-7 transfected with human CD10 molecules)
  • the proteins recovered from each of the cells were separated by SDS-PADE and transferred to a PVDF membrane.
  • Pierce Fast Blocking Buffer Pierce Biotechnology Inc., Tokyo
  • one of the two membranes is with the NEP1 antibody and the other with the anti-CD10 antibody (NCL-CD10, Leica Microsystems, Tokyo)
  • NEP1 antibody the anti-CD10 antibody
  • NCL-CD10 Anti-CD10
  • NEP1 antibody has been internationally deposited as Accession No. NITE BP-02489 at the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation, Patent Microorganisms Depositary Center (Kisasada, Chiba Prefecture 2-5-8-122) (acceptance date: 2017) 12), and is a mouse anti-human monoclonal antibody produced from the hybridoma.
  • Example 2 FACS analysis Control mouse Ig G1 antibody (clone name MOPC) as a primary antibody against HuH-7 cells and their CD10 transfectants (HuH-7 cells transfected with human CD10 molecules) -21, TONBO bioscience San Diego, California), NEP1 antibody, anti-CD10 antibody (clone name B-E3, GEN-PROBE, San Diego, CA), positive control anti-HLA-A, B, C antibody (clone name W6 / 32) , abcam Cambridge, UK) for 30 minutes under ice cooling. Reaction was performed for 30 minutes under ice-cooling using Alexa 647-conjugated rat anti-mouse IgG as a secondary antibody. It analyzed by BD LSRFortessa cell analyzer (BD Bioscience).
  • the results of FACS analysis are shown in FIG.
  • the NEP1 antibody did not bind to HuH-7, but bound to cells transfecting HuH-7 with CD10. This indicates that the NEP1 antibody binds to the CD10 molecule on the cell surface.
  • Example 3 Tumor-bearing mouse treatment experiment NCI-H226 cells (malignant mesothelioma cells, obtained from American Type Culture Collection (ATCC) (Manassas, Virginia)) at 8 x 10 per BALB / c nu mouse Five injections were injected into the abdominal cavity. From 9 days on, administration of 100 ⁇ g / ml of NEP1 antibody per mouse or PBS (control group) was started, and administration was continued at intervals (every 2 to 3 days) shown in FIG. Starting from 7 animals in each group, survival curves were determined.
  • ATCC American Type Culture Collection

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Abstract

L'invention concerne un anticorps anti-CD10 présentant un excellent effet thérapeutique sur des tumeurs malignes. L'invention utilise un anticorps anti-CD10 qui possède une séquence d'acides aminés identique aux séquences d'acides aminés de la chaîne lourde CDRs 1-3 et de la chaîne légère CDRs 1-3 d'un anticorps produit par un hybridome représenté par le numéro d'accès NITE BP-02489.
PCT/JP2017/023104 2017-06-22 2017-06-22 Anticorps anti-cd10 WO2018235247A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022119932A1 (fr) * 2020-12-02 2022-06-09 Agilent Technologies, Inc. Anticorps anti-cd10 humains destinés à une utilisation dans des protocoles d'immunohistochimie (ihc) pour diagnostiquer un cancer
CN116731181A (zh) * 2023-07-17 2023-09-12 武汉爱博泰克生物科技有限公司 抗人cd10蛋白的兔单克隆抗体及其应用
WO2024040194A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BURNOR KJ ET AL.: "Expression of renal cell carcinoma-associated markers erythropoietin, CD 10, and renal cell carcinoma marker in diffuse malignant mesothelioma and metastatic renal cell carcinoma.", ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE, vol. 130, no. 6, 2006, pages 823 - 827, XP055557659 *
MIZUTANI N. ET AL.: "Establishment of anti- mesothelioma monoclonal antibodies.", BMC RESEARCH NOTES, vol. 9, no. 324, 2016, XP055557662 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022119932A1 (fr) * 2020-12-02 2022-06-09 Agilent Technologies, Inc. Anticorps anti-cd10 humains destinés à une utilisation dans des protocoles d'immunohistochimie (ihc) pour diagnostiquer un cancer
WO2024040194A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo
WO2024040195A1 (fr) 2022-08-17 2024-02-22 Capstan Therapeutics, Inc. Conditionnement pour l'ingénierie de cellules immunitaires in vivo
CN116731181A (zh) * 2023-07-17 2023-09-12 武汉爱博泰克生物科技有限公司 抗人cd10蛋白的兔单克隆抗体及其应用
CN116731181B (zh) * 2023-07-17 2024-01-12 武汉爱博泰克生物科技有限公司 抗人cd10蛋白的兔单克隆抗体及其应用

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