WO2021157601A1 - がんを有する対象においてがんを処置することに用いるための抗meflin抗体、および当該抗体を含む医薬組成物 - Google Patents
がんを有する対象においてがんを処置することに用いるための抗meflin抗体、および当該抗体を含む医薬組成物 Download PDFInfo
- Publication number
- WO2021157601A1 WO2021157601A1 PCT/JP2021/003851 JP2021003851W WO2021157601A1 WO 2021157601 A1 WO2021157601 A1 WO 2021157601A1 JP 2021003851 W JP2021003851 W JP 2021003851W WO 2021157601 A1 WO2021157601 A1 WO 2021157601A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- set forth
- antibody
- heavy chain
- light chain
- Prior art date
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 231
- 201000011510 cancer Diseases 0.000 title claims abstract description 129
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 46
- 229940049595 antibody-drug conjugate Drugs 0.000 claims abstract description 122
- 239000000611 antibody drug conjugate Substances 0.000 claims abstract description 114
- 229940127089 cytotoxic agent Drugs 0.000 claims abstract description 22
- 239000002254 cytotoxic agent Substances 0.000 claims abstract description 20
- 231100000599 cytotoxic agent Toxicity 0.000 claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims description 180
- 102000004169 proteins and genes Human genes 0.000 claims description 175
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 30
- 230000000694 effects Effects 0.000 claims description 28
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 22
- 206010039491 Sarcoma Diseases 0.000 claims description 22
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 22
- 201000002528 pancreatic cancer Diseases 0.000 claims description 22
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 22
- 108090000625 Cathepsin K Proteins 0.000 claims description 19
- 102000004171 Cathepsin K Human genes 0.000 claims description 19
- 201000008968 osteosarcoma Diseases 0.000 claims description 17
- 201000009410 rhabdomyosarcoma Diseases 0.000 claims description 14
- 206010033128 Ovarian cancer Diseases 0.000 claims description 13
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 13
- 239000003814 drug Substances 0.000 claims description 12
- 206010006187 Breast cancer Diseases 0.000 claims description 11
- 208000026310 Breast neoplasm Diseases 0.000 claims description 11
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 11
- 206010017758 gastric cancer Diseases 0.000 claims description 11
- 201000011549 stomach cancer Diseases 0.000 claims description 11
- 206010005003 Bladder cancer Diseases 0.000 claims description 10
- 206010009944 Colon cancer Diseases 0.000 claims description 10
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 10
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 10
- 206010029260 Neuroblastoma Diseases 0.000 claims description 10
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 10
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 10
- 208000006990 cholangiocarcinoma Diseases 0.000 claims description 10
- 201000004101 esophageal cancer Diseases 0.000 claims description 10
- 201000005202 lung cancer Diseases 0.000 claims description 10
- 208000020816 lung neoplasm Diseases 0.000 claims description 10
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 9
- 229960002173 citrulline Drugs 0.000 claims description 8
- 201000009030 Carcinoma Diseases 0.000 claims description 6
- 238000003776 cleavage reaction Methods 0.000 claims description 5
- 208000029742 colonic neoplasm Diseases 0.000 claims description 5
- 230000007017 scission Effects 0.000 claims description 5
- 206010004593 Bile duct cancer Diseases 0.000 claims description 4
- 208000026900 bile duct neoplasm Diseases 0.000 claims description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 3
- 206010073140 Clear cell sarcoma of soft tissue Diseases 0.000 claims description 3
- 208000006168 Ewing Sarcoma Diseases 0.000 claims description 3
- 201000008808 Fibrosarcoma Diseases 0.000 claims description 3
- 208000006644 Malignant Fibrous Histiocytoma Diseases 0.000 claims description 3
- 208000031839 Peripheral nerve sheath tumour malignant Diseases 0.000 claims description 3
- 208000015778 Undifferentiated pleomorphic sarcoma Diseases 0.000 claims description 3
- 201000000292 clear cell sarcoma Diseases 0.000 claims description 3
- 206010024627 liposarcoma Diseases 0.000 claims description 3
- 201000009020 malignant peripheral nerve sheath tumor Diseases 0.000 claims description 3
- 208000029974 neurofibrosarcoma Diseases 0.000 claims description 3
- 206010061692 Benign muscle neoplasm Diseases 0.000 claims description 2
- 108010016626 Dipeptides Proteins 0.000 claims description 2
- 201000004458 Myoma Diseases 0.000 claims description 2
- 206010042863 synovial sarcoma Diseases 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 200
- 235000018102 proteins Nutrition 0.000 description 141
- 210000001519 tissue Anatomy 0.000 description 66
- 101000977638 Homo sapiens Immunoglobulin superfamily containing leucine-rich repeat protein Proteins 0.000 description 46
- 102000057179 human ISLR Human genes 0.000 description 45
- 230000000259 anti-tumor effect Effects 0.000 description 39
- 241000699666 Mus <mouse, genus> Species 0.000 description 34
- 230000014509 gene expression Effects 0.000 description 33
- 101100126332 Mus musculus Islr gene Proteins 0.000 description 27
- 238000007901 in situ hybridization Methods 0.000 description 25
- 238000000034 method Methods 0.000 description 25
- 238000002054 transplantation Methods 0.000 description 23
- 238000010186 staining Methods 0.000 description 22
- 239000000427 antigen Substances 0.000 description 17
- 108091007433 antigens Proteins 0.000 description 17
- 102000036639 antigens Human genes 0.000 description 17
- 238000007920 subcutaneous administration Methods 0.000 description 17
- 241000699670 Mus sp. Species 0.000 description 15
- 239000012634 fragment Substances 0.000 description 15
- 210000002536 stromal cell Anatomy 0.000 description 15
- 230000037396 body weight Effects 0.000 description 14
- 210000002950 fibroblast Anatomy 0.000 description 14
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 13
- 238000001727 in vivo Methods 0.000 description 13
- 235000001014 amino acid Nutrition 0.000 description 11
- 231100000433 cytotoxic Toxicity 0.000 description 11
- 230000001472 cytotoxic effect Effects 0.000 description 11
- 230000003834 intracellular effect Effects 0.000 description 11
- 230000003013 cytotoxicity Effects 0.000 description 10
- 231100000135 cytotoxicity Toxicity 0.000 description 10
- 238000011532 immunohistochemical staining Methods 0.000 description 10
- NLMBVBUNULOTNS-HOKPPMCLSA-N [4-[[(2s)-5-(carbamoylamino)-2-[[(2s)-2-[6-(2,5-dioxopyrrol-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl n-[(2s)-1-[[(2s)-1-[[(3r,4s,5s)-1-[(2s)-2-[(1r,2r)-3-[[(1s,2r)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-o Chemical compound C1([C@H](O)[C@@H](C)NC(=O)[C@H](C)[C@@H](OC)[C@@H]2CCCN2C(=O)C[C@H]([C@H]([C@@H](C)CC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCC=2C=CC(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN3C(C=CC3=O)=O)C(C)C)=CC=2)C(C)C)OC)=CC=CC=C1 NLMBVBUNULOTNS-HOKPPMCLSA-N 0.000 description 9
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 9
- 125000003396 thiol group Chemical group [H]S* 0.000 description 9
- 238000011789 NOD SCID mouse Methods 0.000 description 8
- 239000000562 conjugate Substances 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 238000012137 double-staining Methods 0.000 description 8
- 210000004408 hybridoma Anatomy 0.000 description 8
- 229920001223 polyethylene glycol Polymers 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 7
- 238000003556 assay Methods 0.000 description 7
- 201000010099 disease Diseases 0.000 description 7
- -1 radioisotopes Substances 0.000 description 7
- 125000006850 spacer group Chemical group 0.000 description 7
- 238000001262 western blot Methods 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 239000002246 antineoplastic agent Substances 0.000 description 6
- 229910052805 deuterium Inorganic materials 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 210000004072 lung Anatomy 0.000 description 6
- 210000000496 pancreas Anatomy 0.000 description 6
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 5
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 5
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 description 5
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 5
- 239000002033 PVDF binder Substances 0.000 description 5
- 239000002202 Polyethylene glycol Substances 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- 230000004663 cell proliferation Effects 0.000 description 5
- 238000010166 immunofluorescence Methods 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical compound [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 4
- 241000699660 Mus musculus Species 0.000 description 4
- 239000004365 Protease Substances 0.000 description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 description 4
- 230000001605 fetal effect Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 210000003734 kidney Anatomy 0.000 description 4
- 108010093470 monomethyl auristatin E Proteins 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 230000010473 stable expression Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 108010006444 Leucine-Rich Repeat Proteins Proteins 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- 150000001408 amides Chemical group 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 210000003855 cell nucleus Anatomy 0.000 description 3
- 230000001461 cytolytic effect Effects 0.000 description 3
- 238000002073 fluorescence micrograph Methods 0.000 description 3
- 229930004094 glycosylphosphatidylinositol Natural products 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 210000004901 leucine-rich repeat Anatomy 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 2
- 241000699802 Cricetulus griseus Species 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 101150064122 ISLR gene Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 201000009036 biliary tract cancer Diseases 0.000 description 2
- 208000020790 biliary tract neoplasm Diseases 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 229940127093 camptothecin Drugs 0.000 description 2
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 230000005713 exacerbation Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000003125 immunofluorescent labeling Methods 0.000 description 2
- 230000005917 in vivo anti-tumor Effects 0.000 description 2
- 238000001638 lipofection Methods 0.000 description 2
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 208000037819 metastatic cancer Diseases 0.000 description 2
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 208000010125 myocardial infarction Diseases 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- IDBIFFKSXLYUOT-UHFFFAOYSA-N netropsin Chemical compound C1=C(C(=O)NCCC(N)=N)N(C)C=C1NC(=O)C1=CC(NC(=O)CN=C(N)N)=CN1C IDBIFFKSXLYUOT-UHFFFAOYSA-N 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 210000003668 pericyte Anatomy 0.000 description 2
- 230000008823 permeabilization Effects 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000004393 prognosis Methods 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012174 single-cell RNA sequencing Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 230000017423 tissue regeneration Effects 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 108700012359 toxins Proteins 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 229960003048 vinblastine Drugs 0.000 description 2
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- WOWDZACBATWTAU-FEFUEGSOSA-N (2s)-2-[[(2s)-2-(dimethylamino)-3-methylbutanoyl]amino]-n-[(3r,4s,5s)-1-[(2s)-2-[(1r,2r)-3-[[(1s,2r)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-n,3-dimethylbutanamide Chemical compound CC(C)[C@H](N(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)C1=CC=CC=C1 WOWDZACBATWTAU-FEFUEGSOSA-N 0.000 description 1
- INAUWOVKEZHHDM-PEDBPRJASA-N (7s,9s)-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-7-[(2r,4s,5s,6s)-5-hydroxy-6-methyl-4-morpholin-4-yloxan-2-yl]oxy-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound Cl.N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1 INAUWOVKEZHHDM-PEDBPRJASA-N 0.000 description 1
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- SVUOLADPCWQTTE-UHFFFAOYSA-N 1h-1,2-benzodiazepine Chemical compound N1N=CC=CC2=CC=CC=C12 SVUOLADPCWQTTE-UHFFFAOYSA-N 0.000 description 1
- SLAMLWHELXOEJZ-UHFFFAOYSA-N 2-nitrobenzoic acid Chemical compound OC(=O)C1=CC=CC=C1[N+]([O-])=O SLAMLWHELXOEJZ-UHFFFAOYSA-N 0.000 description 1
- 102100022464 5'-nucleotidase Human genes 0.000 description 1
- LGZKGOGODCLQHG-CYBMUJFWSA-N 5-[(2r)-2-hydroxy-2-(3,4,5-trimethoxyphenyl)ethyl]-2-methoxyphenol Chemical compound C1=C(O)C(OC)=CC=C1C[C@@H](O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-CYBMUJFWSA-N 0.000 description 1
- GANZODCWZFAEGN-UHFFFAOYSA-N 5-mercapto-2-nitro-benzoic acid Chemical compound OC(=O)C1=CC(S)=CC=C1[N+]([O-])=O GANZODCWZFAEGN-UHFFFAOYSA-N 0.000 description 1
- VGGWNGWXGFWLRK-UHFFFAOYSA-N 8,9-dihydro-1H-[1,3]oxazolo[4,5-i][1,2]benzodiazepine Chemical compound C1=CC=NNC2=C(OCN3)C3=CC=C21 VGGWNGWXGFWLRK-UHFFFAOYSA-N 0.000 description 1
- DALMAZHDNFCDRP-VMPREFPWSA-N 9h-fluoren-9-ylmethyl n-[(2s)-1-[[(2s)-5-(carbamoylamino)-1-[4-(hydroxymethyl)anilino]-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamate Chemical compound O=C([C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(C)C)NC1=CC=C(CO)C=C1 DALMAZHDNFCDRP-VMPREFPWSA-N 0.000 description 1
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 description 1
- 201000003076 Angiosarcoma Diseases 0.000 description 1
- 108010032595 Antibody Binding Sites Proteins 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 1
- 108090000712 Cathepsin B Proteins 0.000 description 1
- 102000004225 Cathepsin B Human genes 0.000 description 1
- 102000005600 Cathepsins Human genes 0.000 description 1
- 108010084457 Cathepsins Proteins 0.000 description 1
- 102100023126 Cell surface glycoprotein MUC18 Human genes 0.000 description 1
- 208000005243 Chondrosarcoma Diseases 0.000 description 1
- 229930188224 Cryptophycin Natural products 0.000 description 1
- 238000000018 DNA microarray Methods 0.000 description 1
- WDJUZGPOPHTGOT-OAXVISGBSA-N Digitoxin Natural products O([C@H]1[C@@H](C)O[C@@H](O[C@@H]2C[C@@H]3[C@@](C)([C@@H]4[C@H]([C@]5(O)[C@@](C)([C@H](C6=CC(=O)OC6)CC5)CC4)CC3)CC2)C[C@H]1O)[C@H]1O[C@@H](C)[C@H](O[C@H]2O[C@@H](C)[C@@H](O)[C@@H](O)C2)[C@@H](O)C1 WDJUZGPOPHTGOT-OAXVISGBSA-N 0.000 description 1
- 102000016607 Diphtheria Toxin Human genes 0.000 description 1
- 108010053187 Diphtheria Toxin Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 102100037241 Endoglin Human genes 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229940121710 HMGCoA reductase inhibitor Drugs 0.000 description 1
- 208000000435 Heart Rupture Diseases 0.000 description 1
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 description 1
- 101000623903 Homo sapiens Cell surface glycoprotein MUC18 Proteins 0.000 description 1
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 1
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 1
- 101000801254 Homo sapiens Tumor necrosis factor receptor superfamily member 16 Proteins 0.000 description 1
- 229940076838 Immune checkpoint inhibitor Drugs 0.000 description 1
- 102000037984 Inhibitory immune checkpoint proteins Human genes 0.000 description 1
- 108091008026 Inhibitory immune checkpoint proteins Proteins 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 208000032271 Malignant tumor of penis Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- 108010042309 Netropsin Proteins 0.000 description 1
- KYRVNWMVYQXFEU-UHFFFAOYSA-N Nocodazole Chemical compound C1=C2NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=CS1 KYRVNWMVYQXFEU-UHFFFAOYSA-N 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 208000002471 Penile Neoplasms Diseases 0.000 description 1
- 206010034299 Penile cancer Diseases 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 108010084592 Saporins Proteins 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 206010068771 Soft tissue neoplasm Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 208000024313 Testicular Neoplasms Diseases 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 102100033725 Tumor necrosis factor receptor superfamily member 16 Human genes 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 229940122803 Vinca alkaloid Drugs 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- LNHWXBUNXOXMRL-VWLOTQADSA-N belotecan Chemical compound C1=CC=C2C(CCNC(C)C)=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 LNHWXBUNXOXMRL-VWLOTQADSA-N 0.000 description 1
- 229950011276 belotecan Drugs 0.000 description 1
- 229940049706 benzodiazepine Drugs 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000000981 bystander Effects 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- LGZKGOGODCLQHG-UHFFFAOYSA-N combretastatin Natural products C1=C(O)C(OC)=CC=C1CC(O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-UHFFFAOYSA-N 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 108010006226 cryptophycin Proteins 0.000 description 1
- PSNOPSMXOBPNNV-VVCTWANISA-N cryptophycin 1 Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H]2[C@H](O2)C=2C=CC=CC=2)C/C=C/C(=O)N1 PSNOPSMXOBPNNV-VVCTWANISA-N 0.000 description 1
- PSNOPSMXOBPNNV-UHFFFAOYSA-N cryptophycin-327 Natural products C1=C(Cl)C(OC)=CC=C1CC1C(=O)NCC(C)C(=O)OC(CC(C)C)C(=O)OC(C(C)C2C(O2)C=2C=CC=CC=2)CC=CC(=O)N1 PSNOPSMXOBPNNV-UHFFFAOYSA-N 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 208000024558 digestive system cancer Diseases 0.000 description 1
- WDJUZGPOPHTGOT-XUDUSOBPSA-N digitoxin Chemical compound C1[C@H](O)[C@H](O)[C@@H](C)O[C@H]1O[C@@H]1[C@@H](C)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@@H]3C[C@@H]4[C@]([C@@H]5[C@H]([C@]6(CC[C@@H]([C@@]6(C)CC5)C=5COC(=O)C=5)O)CC4)(C)CC3)C[C@@H]2O)C)C[C@@H]1O WDJUZGPOPHTGOT-XUDUSOBPSA-N 0.000 description 1
- 229960000648 digitoxin Drugs 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 201000010231 gastrointestinal system cancer Diseases 0.000 description 1
- UIVFUQKYVFCEKJ-OPTOVBNMSA-N gimatecan Chemical compound C1=CC=C2C(\C=N\OC(C)(C)C)=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UIVFUQKYVFCEKJ-OPTOVBNMSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 239000012274 immune-checkpoint protein inhibitor Substances 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 230000002601 intratumoral effect Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000007914 intraventricular administration Methods 0.000 description 1
- 229960004768 irinotecan Drugs 0.000 description 1
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 208000012804 lymphangiosarcoma Diseases 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 206010027191 meningioma Diseases 0.000 description 1
- 229960001156 mitoxantrone Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 210000000651 myofibroblast Anatomy 0.000 description 1
- 229950006344 nocodazole Drugs 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 210000002705 pancreatic stellate cell Anatomy 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 210000004180 plasmocyte Anatomy 0.000 description 1
- 229930001119 polyketide Natural products 0.000 description 1
- 150000003881 polyketide derivatives Chemical class 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- YUOCYTRGANSSRY-UHFFFAOYSA-N pyrrolo[2,3-i][1,2]benzodiazepine Chemical compound C1=CN=NC2=C3C=CN=C3C=CC2=C1 YUOCYTRGANSSRY-UHFFFAOYSA-N 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 210000001525 retina Anatomy 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000002363 skeletal muscle cell Anatomy 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 238000011706 wistar kyoto rat Methods 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
- A61K47/68031—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being an auristatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/77—Internalization into the cell
Definitions
- the present invention relates to an anti-MEFLIN antibody for use in treating cancer in a subject having cancer, and a pharmaceutical composition containing the antibody.
- MEFLIN is a leucine-rich repeat (LRR) encoded by the ISLR (Immunoglobulin superior-rich repeat) gene obtained from a library of human genes that are predominantly expressed in the retina as compared to the brain. It is a membrane protein containing a (Ig) -like domain (Non-Patent Documents 1 and 2). It is also known by the ISLR name similar to the gene name (Non-Patent Document 1). MEFLIN exists bound to the cell membrane surface by a GPI (glycosylphosphatidylinositol) anchor, but it has also been reported that it is cleaved near the cell membrane and secreted extracellularly (Non-Patent Document 2).
- GPI glycosepiderglycosylphosphatidylinositol
- MEFLIN is a protein specifically expressed in mesenchymal stem cells (Non-Patent Document 2). Many molecules such as CD105, CD73, CD90, CD146, and CD271 are known as markers for mesenchymal stem cells, and MEFLIN is reported to be the most specific marker molecule for mesenchymal stem cells. (Non-Patent Document 2, Patent Document 1).
- Non-Patent Document 2 Mesenchymal stem cells expressing MEFLIN are abundant around blood vessels in all organs or in connective tissue, and are present in osteoblasts, chondroblasts, lipoblasts, skeletal muscle cells, myofibroblasts, and nerve cells. Has the ability to differentiate into (Non-Patent Document 2). There is also a report that mesenchymal stem cells are almost the same cells as pericytes (vascular pericytes) and fibroblasts around blood vessels (Non-Patent Document 3).
- Non-Patent Document 4 fibroblasts derived from mesenchymal stem cells are known to proliferate around the cancer cells and are called cancer-associated fibroblasts (or CAF) (Non-Patent Document 4).
- Cancer-related fibroblasts are observed in tissues of almost all cancer types, but are known to show remarkable proliferation especially in intractable cancers such as pancreatic cancer, cholangiocarcinoma, breast cancer, and poorly differentiated gastrointestinal tract cancer (non-cancer).
- Patent Documents 4 and 5 MEFLIN is positive for cancer-related fibroblasts, which can be examined by in situ hybridization for detecting mRNA derived from the ISLR gene or immunohistochemical staining using an antibody (Non-Patent Document 5).
- Biopsy materials or surgical materials derived from cancer patients are used for in situ hybridization and immunohistochemical staining.
- MEFLIN is known to be specifically expressed in cancer-related fibroblasts in cancer tissues, and is not expressed in cancer cells, vascular endothelial cells, smooth muscle cells, blood cell lineage cells, and nerve cells (Non-Patent Documents). 5).
- Non-Patent Documents 5 and 2 the pancreatic cancer patient group in which MEFLIN-positive cancer-related fibroblasts account for 20% or more of all cancer-related fibroblasts shows a better prognosis than the patient group in which less than 20%. It has also been reported that the therapeutic effect (response rate) of immune checkpoint inhibitors is low in a group of patients in which MEFLIN-positive cancer-related fibroblasts account for less than 20% of all cancer-related fibroblasts (Patent Document 2).
- MEFLIN-positive mesenchymal stem cells or fibroblasts are known to be important cells for tissue repair of many organs. For example, in a mouse myocardial infarction model, a high degree of accumulation of MEFLIN-positive cells is observed in the acute phase after myocardial infarction (Non-Patent Document 6). Expression of MEFLIN in these fibroblasts is essential for myocardial repair, and cardiac rupture is observed in mice lacking the ISLR gene (Non-Patent Document 6). There is also a report that MEFLIN-positive fibroblasts function suppressively on fibrosis and tissue sclerosis after tissue repair (Non-Patent Document 6).
- the present invention provides an anti-MEFLIN antibody for use in treating cancer in a subject having cancer, and a pharmaceutical composition containing the antibody.
- the cancer can be a sarcoma and a MEFLIN-negative cancer.
- the present inventors have found that an antibody-drug conjugate of an antibody that binds to the MEFLIN protein and a cytotoxic agent exerts an antitumor effect on sarcoma.
- the present invention is based on these findings.
- a pharmaceutical composition for use in treating cancer which comprises an antibody-drug conjugate (ADC) of an antibody that binds to MEFLIN and a cytotoxic agent.
- ADC antibody-drug conjugate
- the ADC is an ADC in which an antibody and a drug are linked via a linker, and the linker has a cleavage site that is cleaved inside the cell. ..
- An antibody that binds to MEFLIN is selected from the group consisting of the following: (1A) Variable heavy chain including heavy chain CDR1 having the amino acid sequence set forth in SEQ ID NO: 1, heavy chain CDR2 having the amino acid sequence set forth in SEQ ID NO: 2, and heavy chain CDR3 having the amino acid sequence set forth in SEQ ID NO: 3.
- a light chain variable comprising a region and a light chain CDR1 having the amino acid sequence set forth in SEQ ID NO: 4, a light chain CDR2 having the amino acid sequence set forth in SEQ ID NO: 5, and a light chain CDR3 having the amino acid sequence set forth in SEQ ID NO: 6.
- Antibodies with regions (1B) An antibody having a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region having the amino acid sequence set forth in SEQ ID NO: 8; (1C) An antibody that competes for binding of the antibody of (1B) above and the MEFLIN protein; and (1D) An antibody that binds the antibody of (1B) above and an epitope that overlaps on the MEFLIN protein; (2A) A heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 9, the heavy chain CDR2 set forth in SEQ ID NO: 10, and the heavy chain CDR3 set forth in SEQ ID NO: 11.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 12, the light chain CDR2 set forth in SEQ ID NO: 13, and the light chain CDR3 set forth in SEQ ID NO: 14.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 20, the light chain CDR2 set forth in SEQ ID NO: 21, and the light chain CDR3 set forth in SEQ ID NO: 22;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 28, the light chain CDR2 set forth in SEQ ID NO: 29, and the light chain CDR3 set forth in SEQ ID NO: 30;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 36, the light chain CDR2 set forth in SEQ ID NO: 37, and the light chain CDR3 set forth in SEQ ID NO: 38;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 44, the light chain CDR2 set forth in SEQ ID NO: 45, and the light chain CDR3 set forth in SEQ ID NO: 46;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 52, the light chain CDR2 set forth in SEQ ID NO: 53, and the light chain CDR3 set forth in SEQ ID NO: 54;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 60, the light chain CDR2 set forth in SEQ ID NO: 61, and the light chain CDR3 set forth in SEQ ID NO: 62;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 68, the light chain CDR2 set forth in SEQ ID NO: 69, and the light chain CDR3 set forth in SEQ ID NO: 70;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 76, the light chain CDR2 set forth in SEQ ID NO: 77, and the light chain CDR3 set forth in SEQ ID NO: 78;
- 10B An antibody having the heavy chain variable region set forth in SEQ ID NO: 79 and the light chain variable region set forth in SEQ ID NO: 80;
- 10C Antibodies that compete for binding of the antibody of (10B) above to the MEFLIN protein; and
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 84, the light chain CDR2 set forth in SEQ ID NO: 85, and the light chain CDR3 set forth in SEQ ID NO: 86;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 92, the light chain CDR2 set forth in SEQ ID NO: 93, and the light chain CDR3 set forth in SEQ ID NO: 94; (12B) An antibody having the heavy chain variable region set forth in SEQ ID NO: 95 and the light chain variable region set forth in SEQ ID NO: 96; (12C) Antibodies that compete for binding of the antibody of (12B) above to the MEFLIN protein; and (12D) The antibody of (12B) above and an antibody that binds to an overlapping epitope on the MEFLIN protein; (13A) A heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 97, the heavy chain CDR2 set forth in SEQ ID NO: 98, and the heavy chain CDR3 set forth in SEQ ID NO: 99.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 100, the light chain CDR2 set forth in SEQ ID NO: 101, and the light chain CDR3 set forth in SEQ ID NO: 102;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 108, the light chain CDR2 set forth in SEQ ID NO: 109, and the light chain CDR3 set forth in SEQ ID NO: 110;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 116, the light chain CDR2 set forth in SEQ ID NO: 117, and the light chain CDR3 set forth in SEQ ID NO: 118;
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 124, the light chain CDR2 set forth in SEQ ID NO: 125, and the light chain CDR3 set forth in SEQ ID NO: 126;
- a heavy chain variable region comprising the heavy chain CDR1 described, the heavy chain CDR2 set forth in SEQ ID NO: 130, and the heavy chain CDR3 set forth in SEQ ID NO: 131.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 132, the light chain CDR2 set forth in SEQ ID NO: 133, and the light chain CDR3 set forth in SEQ ID NO: 134;
- (17C) An antibody that competes for binding of the antibody of (17B) above and the MEFLIN protein; and (17D) an antibody that binds the antibody of (17B) above and an overlapping epitope on the MEFLIN protein.
- a pharmaceutical composition comprising an antibody-drug conjugate (ADC) of the antibody according to the above [4] and a cytotoxic agent.
- ADC antibody-drug conjugate
- the pharmaceutical composition according to the above [5] which is used for treating cancer.
- Sarcomas include mucosal fibrosarcoma, malignant fibrous histiocytoma, fatty sarcoma, smooth sarcoma, rhabdomyosarcoma, neuroblastoma, malignant peripheral nerve sheath tumor, Ewing sarcoma, epithelial sarcoma, clear cell sarcoma,
- the cancer is selected from the group consisting of breast cancer, pancreatic cancer, lung cancer, colon cancer, stomach cancer, bile duct cancer, ovarian cancer, bladder cancer, and esophageal cancer, as described above [ 10]
- the pharmaceutical composition according to. [13] The pharmaceutical composition according to any one of the above [10] to [12], wherein the antibody does not have internalizing activity.
- FIG. 1 shows the results of Western blotting showing the binding property between the human MEFLIN protein and the monoclonal antibody obtained from various clones.
- FIG. 1 information on the clone name and the fragment of MEFLIN (the region of the fragment is indicated by the amino acid number) is shown.
- FIG. 2 shows the results of Western blotting showing the binding property between the human MEFLIN protein and the monoclonal antibody obtained from various clones.
- FIG. 2 information on the clone name and the fragment of MEFLIN (the region of the fragment is indicated by the amino acid number) is shown.
- FIG. 3 is a fluorescence microscope image showing the intracellular uptake (internalization) activity of monoclonal antibodies obtained from various clones.
- FIG. 4 is a fluorescence microscope image showing the intracellular uptake (internalization) activity of monoclonal antibodies obtained from various clones. Cell nuclei were stained with DAPI and the antibody was detected with Alexa488-labeled antibody.
- FIG. 5 is a fluorescence microscope image showing the intracellular uptake (internalization) activity of monoclonal antibodies obtained from various clones. Cell nuclei were stained with DAPI and the antibody was detected with Alexa488-labeled antibody.
- FIG. 4 is a fluorescence microscope image showing the intracellular uptake (internalization) activity of monoclonal antibodies obtained from various clones. Cell nuclei were stained with DAPI and the antibody was detected with Alexa488-labeled antibody.
- FIG. 6 shows the in vitro cell proliferation inhibitory activity of the prepared antibody-drug conjugate (ADC) on human MEFLIN protein overexpressing cells (HEK293 cells).
- FIG. 6 also shows the FACS analysis results of the anti-MEFLIN monoclonal antibody-treated group and the untreated group (negative control) against human MEFLIN protein overexpressing cells (HEK293 cells).
- the horizontal axis shows the expression level of MEFLIN protein, and the vertical axis shows the frequency of cells.
- FIG. 7 shows the in vitro cell proliferation inhibitory activity of ADC against human rhabdomyosarcoma cell line (KYM-1 cells).
- FIG. 7 also shows the FACS analysis results of the anti-MEFLIN monoclonal antibody-treated group and the untreated group (negative control) against the human rhabdomyosarcoma cell line (KYM-1 cells) used.
- the horizontal axis shows the expression level of MEFLIN protein, and the vertical axis shows the frequency of cells.
- FIG. 8 shows the in vivo antitumor effect of ADC on a cancer-bearing model mouse subcutaneously transplanted with a rhabdomyosarcoma cell line (KYM-1). The arrows in the graph indicate the timing of administration.
- FIG. 8 also shows the expression of human MEFLIN in transplanted rhabdomyosarcoma tissue.
- FIG. 9 shows the in vivo antitumor effect of ADC on a cancer-bearing model mouse subcutaneously transplanted with an osteosarcoma cell line (HsOs1). The arrows in the graph indicate the timing of administration.
- FIG. 9 also shows the expression of human MEFLIN in transplanted osteosarcoma tissue.
- FIG. 10 shows the antitumor effect of ADC in vivo on a cancer-bearing model mouse subcutaneously transplanted with a pancreatic cancer cell line (BxPC-3). The arrows in the graph indicate the timing of administration.
- FIG. 10 also shows the expression of mouse MEFLIN in transplanted pancreatic cancer tissues and stroma.
- FIG. 11 shows the antitumor effect of ADC in vivo on a cancer-bearing model mouse subcutaneously transplanted with a lung cancer cell line (A549). The arrows in the graph indicate the timing of administration.
- FIG. 11 also shows the expression of mouse MEFLIN in transplanted lung cancer tissues and stroma.
- FIG. 12 shows the antitumor effect of ADC in vivo on a cancer-bearing model mouse subcutaneously transplanted with a neuroblastoma cell line (NB-1). The arrows in the graph indicate the timing of administration.
- FIG. 12 also shows the expression of human MEFLIN in transplanted neuroblastoma tissue.
- FIG. 13 shows the antitumor effect of ADC in vivo on a cancer-bearing model mouse subcutaneously transplanted with a colorectal cancer cell line (DLD-1). The arrows in the graph indicate the timing of administration.
- FIG. 13 also shows the expression of mouse MEFLIN in transplanted colorectal cancer tissues and stroma.
- FIG. 14 shows the antitumor effect of ADC in vivo on a cancer-bearing model mouse subcutaneously transplanted with a gastric cancer cell line (MKN45). The arrows in the graph indicate the timing of administration.
- FIG. 14 also shows the expression of mouse MEFLIN in transplanted gastric cancer tissues and stroma.
- FIG. 15 shows the results of analysis of single-cell RNA sequencing data (Tabula Muris) using mouse pancreas deposited on the Internet. It is shown that the MEFLIN-positive cell population and the cathepsin K-positive cell population in the mouse pancreas match (arrows).
- FIG. 16 shows the results of analysis of single-cell RNA sequencing data (Tabula Muris) using mouse lungs deposited on the Internet. It is shown that the MEFLIN-positive cell population and the cathepsin K-positive cell population in the mouse lung match (arrows).
- FIG. 17 shows the expression of cathepsin K in an exogenous mouse MEFLIN-expressing CHO cell line.
- FIG. 18 shows the results of immunofluorescence double staining using an anti-MEFLIN monoclonal antibody and an anti-cathepsin K antibody on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an osteosarcoma cell line (HsOs1). .. It indicates that cathepsin K (red) is secreted around MEFLIN-positive cells (green) (arrow).
- FIG. 19 immunofluorescence double staining using an anti-MEFLIN monoclonal antibody and an anti-cathepsin K antibody was performed on the tumor tissue of a cancer-bearing model mouse in which a pancreatic cancer cell line (BxPC-3) was skin-transplanted. The results are shown.
- FIG. 20 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with a cholangiocarcinoma cell line (HuCCT1). The arrows in the graph indicate the timing of administration.
- FIG. 20 also shows the expression of mouse MEFLIN in transplanted cholangiocarcinoma tissues and interstitium.
- FIG. 21 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with a bladder cancer cell line (T24). The arrows in the graph indicate the timing of administration.
- FIG. 21 also shows the expression of mouse MEFLIN in transplanted bladder cancer tissues and stroma.
- FIG. 22 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an ovarian cancer cell line (OV-90). The arrows in the graph indicate the timing of administration.
- FIG. 22 also shows the expression of mouse MEFLIN in transplanted ovarian cancer tissues and stroma.
- FIG. 23 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an esophageal cancer cell line (KYSE). The arrows in the graph indicate the timing of administration.
- FIG. 22 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an esophageal cancer cell line (KYSE). The arrows in the graph indicate the timing of administration.
- FIG. 23 also shows the expression of mouse MEFLIN in transplanted esophageal cancer tissues and stroma.
- FIG. 24 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an osteosarcoma cell line (HsOs1). The arrows in the graph indicate the timing of administration.
- FIG. 25 shows the antitumor effect of ADC in vivo on the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with a breast cancer cell line (MCF7). The arrows in the graph indicate the timing of administration.
- FIG. 25 also shows the expression of mouse MEFLIN in transplanted breast cancer tissues and stroma.
- FIG. 26 shows the results of subjecting a tissue section of a human pancreatic cancer surgical specimen to immunohistology (IHC) staining using an anti-MEFLIN antibody, and the result of staining by in situ hybridization (ISH).
- IHC immunohistology
- ISH in situ hybridization
- the "object” means a mammal, and may be a human in particular.
- treatment is used in the sense of including treatment (therapeutic treatment) and prevention (preventive treatment).
- treatment means the treatment, cure, prevention, improvement of remission, or reduction of the rate of progression of a disease or disorder.
- prevention means reducing the likelihood of developing a disease or condition, or delaying the onset of a disease or condition.
- disease means a symptomatology for which treatment is beneficial.
- cancer means a malignant tumor.
- antibody means immunoglobulin and includes polyclonal antibody and monoclonal antibody.
- Preferred antibodies are monoclonal antibodies.
- the origin of the antibody is not particularly limited, and examples thereof include non-human animal antibody, non-human mammalian antibody, and human antibody.
- the antibody may be a chimeric antibody, a humanized antibody, or a human antibody.
- the antibody may be a bispecific antibody.
- the antibody used as a medicine can be preferably a chimeric antibody, more preferably a humanized antibody, and even more preferably a human antibody.
- the bispecific antibody can be a monoclonal antibody, preferably a chimeric antibody, more preferably a humanized antibody, and even more preferably a human antibody.
- the "therapeutically effective amount” means the amount of a drug effective for treating (preventing or treating) a disease or condition.
- a therapeutically effective amount of a drug slows the rate of exacerbation of a symptom of a disease or condition, stops the exacerbation of the symptom, ameliorates the symptom, cures the symptom, or develops or develops the symptom. It is possible to suppress.
- “competing” means competing for binding to another binding antibody with respect to binding to an antigen. Competition can occur when two antibody binding sites overlap for an antigen.
- Such antibodies can be obtained by immunization with the epitope as described above, and / or by competing assays to determine if the binding of one antibody to the antigen is reduced by the other antibody. Obtainable.
- the "antibody-drug conjugate” (hereinafter, also referred to as "ADC") means a substance in which an antibody and a cytotoxic agent are linked.
- ADC the antibody and the cytotoxic agent can be linked via a suitable linker.
- cytotoxic agents chemotherapeutic agents, radioisotopes, and toxins can be used.
- the ADC also includes a conjugate of the antigen-binding fragment of the antibody with the drug.
- the term "antigen-binding fragment” means a part of an antibody that maintains its binding property to an antigen.
- the antigen-binding fragment may contain a heavy chain variable region, a light chain variable region, or both of the antibodies of the present invention.
- the antigen-binding fragment may be chimeric or humanized.
- Examples of the antigen-binding fragment include Fab, Fab', F (ab') 2 , Fv, scFv (single chain Fv), diabody, sc (Fv) 2 (single chain (Fv) 2 ).
- Fragments of such antibodies are not particularly limited, but can be obtained, for example, by treating the antibody with an enzyme.
- Fab can be obtained by digesting the antibody with papain.
- digestion of the antibody with pepsin can give F (ab') 2
- further reduction can give Fab'.
- an antigen-binding fragment of such an antibody can be used.
- MEFLIN or “Meflin” is a protein also called a leucine-rich repeat-containing immunoglobulin superfamily (ISLR).
- Human MEFLIN may have an amino acid sequence registered under GenBank Accession No. BAA859070.1.
- MEFLIN can be a MEFLIN having an amino acid sequence corresponding to the amino acid sequence registered under GenBank Accession No. BAA859070.1 (eg, human MEFLIN. If you want to identify the animal species from which it is derived, then MEFLIN Alternatively, it is expressed as hMEFLIN), mouse MEFLIN (mMEFLIN), or the like.
- the antibody and the cytotoxic agent are linked via a linker.
- a chemotherapeutic agent for example, an anticancer agent such as a commercially available anticancer agent, for example, oristatin (auristatin E, oristatin F phenylenediamine (AFP), monomethyloristatin E, monomethylori) Statin F and its derivatives), maytancinoids DM1 and DM4 and their derivatives), camptothecin (SN-38, irinotecan, roottecan, DB67, BMP1350, ST1481, CKD602, topotecan and exotecin, and their derivatives), DNA sub Groove-binding agents (engine, lexitropcin, duocalmycin and their derivatives), taxan (pacrytaxel and docetaxel and their derivatives), polyketide (discodelmolide and its derivatives), anthraquinone
- an anticancer agent such as a commercially available anticancer agent, for example,
- cytotoxic agent in the ADC of the present invention for example, camptothecin, particularly SN-38, or execan can be used. Any cytotoxic agent that is used for the treatment of cancer can be used.
- cytotoxic agent a pharmaceutically acceptable salt, solvate (for example, hydrate), ester, or prodrug of the cytotoxic agent may be used.
- the ADC linker can be a non-cleavable linker or a cleavable linker. Such a linker can be appropriately selected and synthesized by those skilled in the art in the preparation of the ADC.
- Cleavable linkers also include linkers having a degradable bond such as an ester bond.
- Cleavable linkers include linkers having protease cleavage sites such as cleaving moieties such as valine-citrulline or valine-alanine peptides. The peptide region consisting of valine-citrulline can be cleaved by proteases such as cathepsin B.
- the linker may have a first spacer introduced, for example, between the antibody and the cleaving moiety, eg, polyethylene glycol (PEG), eg, about 5-40 repeat units per molecule. PEG can be used as the first spacer.
- a second spacer may be introduced between the cleaving moiety and the cytotoxic agent, for example p-aminobenzyloxycarbonyl (PABC) can be used as the second spacer.
- PEG polyethylene glycol
- PABC p-aminobenzyloxycarbonyl
- Cleavable linkers are physiologically stable except at the site of cleavage in the cancerous tissue (particularly, they are physiologically stable until they reach the cancerous tissue).
- the linker comprises a first spacer and a cleaving moiety.
- the linker comprises a first spacer, a cleaving moiety, and a second spacer.
- the linker comprises PEG, a cleaving moiety, and PABC.
- the binding of the antibody to the linker can be, for example, linked to the sulfhydryl group of the antibody via a maleimide group.
- the antibody is linked to the anti-cancer agent via its sulfhydryl group with a linker having a maleimide-PEG-cleavable moiety.
- the antibody is linked to the anti-cancer agent via its sulfhydryl group with a linker having maleimide-PEG-cleavable moiety-PABC.
- the ADC may have the structure represented by formula (II) below. The ADC of the present invention is considered to be useful as a therapeutic agent for cancer.
- an antibody having an amino acid sequence of heavy chain CDR1 to 3 and light chain CDR1 to 3 of an antibody produced from a clone selected from the following is provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- an antibody having an amino acid sequence of a heavy chain variable region and a light chain variable region of an antibody produced from a clone selected from the following can be provided.
- the antibody may preferably be a human chimeric antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 1, the heavy chain CDR2 set forth in SEQ ID NO: 2, and the heavy chain CDR3 set forth in SEQ ID NO: 3.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 4, the light chain CDR2 set forth in SEQ ID NO: 5, and the light chain CDR3 set forth in SEQ ID NO: 4;
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 9, the heavy chain CDR2 set forth in SEQ ID NO: 10, and the heavy chain CDR3 set forth in SEQ ID NO: 11.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 12, the light chain CDR2 set forth in SEQ ID NO: 13, and the light chain CDR3 set forth in SEQ ID NO: 14.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 17, the heavy chain CDR2 set forth in SEQ ID NO: 18, and the heavy chain CDR3 set forth in SEQ ID NO: 19.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 20, the light chain CDR2 set forth in SEQ ID NO: 21, and the light chain CDR3 set forth in SEQ ID NO: 22; (3B) An antibody having the heavy chain variable region set forth in SEQ ID NO: 23 and the light chain variable region set forth in SEQ ID NO: 24; (3C) Antibodies that compete for binding of the antibody of (3B) above and the MEFLIN protein; and (3D) Antibodies that bind the antibody of (3B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 25, the heavy chain CDR2 set forth in SEQ ID NO: 26, and the heavy chain CDR3 set forth in SEQ ID NO: 27.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 28, the light chain CDR2 set forth in SEQ ID NO: 29, and the light chain CDR3 set forth in SEQ ID NO: 30;
- (4D) Antibodies that bind the antibody of (4B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- the present invention is an antibody that binds to a MEFLIN protein.
- (5D) Antibodies that bind the antibody of (5B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 41, the heavy chain CDR2 set forth in SEQ ID NO: 42, and the heavy chain CDR3 set forth in SEQ ID NO: 43.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 44, the light chain CDR2 set forth in SEQ ID NO: 45, and the light chain CDR3 set forth in SEQ ID NO: 46;
- (6D) Antibodies that bind the antibody of (6B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 49, the heavy chain CDR2 set forth in SEQ ID NO: 50, and the heavy chain CDR3 set forth in SEQ ID NO: 51.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 52, the light chain CDR2 set forth in SEQ ID NO: 53, and the light chain CDR3 set forth in SEQ ID NO: 54;
- (7D) Antibodies that bind the antibody of (7B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 57, the heavy chain CDR2 set forth in SEQ ID NO: 58, and the heavy chain CDR3 set forth in SEQ ID NO: 59.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 60, the light chain CDR2 set forth in SEQ ID NO: 61, and the light chain CDR3 set forth in SEQ ID NO: 62; (8B) An antibody having the heavy chain variable region set forth in SEQ ID NO: 63 and the light chain variable region set forth in SEQ ID NO: 64; (8C) Antibodies that compete for binding of the antibody of (8B) above and the MEFLIN protein; and (8D) Antibodies that bind the antibody of (8B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 65, the heavy chain CDR2 set forth in SEQ ID NO: 66, and the heavy chain CDR3 set forth in SEQ ID NO: 67.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 68, the light chain CDR2 set forth in SEQ ID NO: 69, and the light chain CDR3 set forth in SEQ ID NO: 70; (9B) An antibody having the heavy chain variable region set forth in SEQ ID NO: 71 and the light chain variable region set forth in SEQ ID NO: 72; (9C) Antibodies that compete for binding of the antibody of (9B) above and the MEFLIN protein; and (9D) Antibodies that bind the antibody of (9B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 73, the heavy chain CDR2 set forth in SEQ ID NO: 74, and the heavy chain CDR3 set forth in SEQ ID NO: 75.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 76, the light chain CDR2 set forth in SEQ ID NO: 77, and the light chain CDR3 set forth in SEQ ID NO: 78;
- (10D) Antibodies that bind the antibody of (10B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 81, the heavy chain CDR2 set forth in SEQ ID NO: 82, and the heavy chain CDR3 set forth in SEQ ID NO: 83.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 84, the light chain CDR2 set forth in SEQ ID NO: 85, and the light chain CDR3 set forth in SEQ ID NO: 86;
- (11D) Antibodies that bind the antibody of (11B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 89, the heavy chain CDR2 set forth in SEQ ID NO: 90, and the heavy chain CDR3 set forth in SEQ ID NO: 91.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 92, the light chain CDR2 set forth in SEQ ID NO: 93, and the light chain CDR3 set forth in SEQ ID NO: 94; (12B) An antibody having the heavy chain variable region set forth in SEQ ID NO: 95 and the light chain variable region set forth in SEQ ID NO: 96; (12C) Antibodies that compete for binding of the antibody of (12B) above and the MEFLIN protein; and (12D) Antibodies that bind the antibody of (12B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 97, the heavy chain CDR2 set forth in SEQ ID NO: 98, and the heavy chain CDR3 set forth in SEQ ID NO: 99.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 100, the light chain CDR2 set forth in SEQ ID NO: 101, and the light chain CDR3 set forth in SEQ ID NO: 102;
- (13D) an antibody that binds the antibody of (13B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 105, the heavy chain CDR2 set forth in SEQ ID NO: 106, and the heavy chain CDR3 set forth in SEQ ID NO: 107.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 108, the light chain CDR2 set forth in SEQ ID NO: 109, and the light chain CDR3 set forth in SEQ ID NO: 110;
- (14D) Antibodies that bind the antibody of (14B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 113, the heavy chain CDR2 set forth in SEQ ID NO: 114, and the heavy chain CDR3 set forth in SEQ ID NO: 115.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 116, the light chain CDR2 set forth in SEQ ID NO: 117, and the light chain CDR3 set forth in SEQ ID NO: 118;
- (15D) Antibodies that bind the antibody of (15B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 121, the heavy chain CDR2 set forth in SEQ ID NO: 122, and the heavy chain CDR3 set forth in SEQ ID NO: 123.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 124, the light chain CDR2 set forth in SEQ ID NO: 125, and the light chain CDR3 set forth in SEQ ID NO: 126;
- (16D) Antibodies that bind the antibody of (16B) above and an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- a heavy chain variable region comprising the heavy chain CDR1 set forth in SEQ ID NO: 129, the heavy chain CDR2 set forth in SEQ ID NO: 130, and the heavy chain CDR3 set forth in SEQ ID NO: 131.
- An antibody having a light chain variable region comprising the light chain CDR1 set forth in SEQ ID NO: 132, the light chain CDR2 set forth in SEQ ID NO: 133, and the light chain CDR3 set forth in SEQ ID NO: 134;
- An antibody that competes for binding of the antibody of (17B) above to the MEFLIN protein; and (17D) an antibody that binds the antibody of (17B) above to an overlapping epitope on the MEFLIN protein are provided.
- the antibody is preferably a human chimeric antibody, preferably a humanized antibody.
- the antibody that binds to the MEFLIN protein binds to the human MEFLIN protein. In some aspects of the invention, the antibody that binds to the MEFLIN protein specifically binds to the human MEFLIN protein.
- specifically binding is meant binding to a MEFLIN protein with a stronger affinity than binding to at least one other protein.
- Specifically binding includes the case of binding to a human MEFLIN protein with a stronger affinity than that of a mouse MEFLIN protein.
- Competing antibodies can be identified by a cross-blocking assay, preferably a competing ELISA assay.
- the antigen is coated, for example, on a microtiter plate, to which the presence of a candidate competing antibody is added and incubated to form a bond between the antigen and the candidate antibody.
- the antibody can be further added to the well, incubated, washed, and the amount of the desired antibody bound can be quantified to determine whether or not the antibody has competed. In case of competition, less labeling should remain in the wells.
- HDX-MS hydrogen-deuterium exchange mass spectrometry
- the exchange of amide protons in protein complexes with deuterium can be detected in the presence of heavy water.
- the amide protons on the interaction surface of the protein complex are less likely to be exchanged for deuterium (or the exchange rate is slower), and the amide protons in the exposed regions are more likely to be exchanged for deuterium (or the exchange rate). Is fast).
- the region where the decrease in the exchange rate of hydrogen to deuterium is observed can be considered as the region that binds to the antibody, thereby the interaction surface on the binding partner of the antibody ( The epitope region) can be determined.
- the epitope region By determining the epitope regions of each of the two antibodies by the above method, it is possible to determine whether or not the epitope regions overlap.
- the exchange of hydrogen to deuterium can be detected by those skilled in the art by mass spectrometry.
- the antibody that binds to the MEFLIN protein is an antibody that has intracellular anatomicalizing activity. In some aspects of the invention, the antibody that binds to the MEFLIN protein is an antibody that does not have intracellular anatomicalizing activity. In one aspect of the invention, the cancer cell is MEFLIN protein positive and the antibody that binds to the MEFLIN protein is an antibody that has intracellular anatomical activity. In one aspect of the invention, the cancer cell is MEFLIN protein positive and the antibody that binds to the MEFLIN protein is an antibody that does not have intracellular anatomicalizing activity.
- the cancer stromal cells are MEFLIN protein positive and the antibody that binds to the MEFLIN protein is an antibody that has intracellular anatomical activity. In one aspect of the invention, the cancer stromal cells are MEFLIN protein positive and the antibody that binds to the MEFLIN protein is an antibody that does not have intracellular anatomicalizing activity.
- the ADC may have a cleaving linker. In these embodiments, the ADC may have a non-cleavable linker. If the payload of the ADC is prodrugized to be cytotoxic for the first time after cleavage of the linker, the linker is preferably a cleaving linker.
- the ADCs of the invention have an internalizing activity and have a cleaving linker that cleaves intracellularly. In some embodiments, the ADCs of the invention have no internalizing activity and have a cleaving linker that cleaves extracellularly.
- Whether or not the antibody has internalizing activity can be confirmed by an in vitro test.
- cells having the MEFLIN protein on the cell surface are contacted with an antibody that binds to the MEFLIN protein, incubated for a sufficient time (for example, about 15 minutes) for the antibody to internalize, and then in the culture medium and in the culture medium. It can be confirmed by removing the antibody bound to the cell surface by washing (for example, an aqueous solution containing 0.5 M NaCl and 3% by volume acetic acid), and then staining the antibody internalized inside the cell.
- Staining of the antibody internalized into cells can be performed in the same manner as staining of intracellular proteins. Staining of the antibody internalized into cells can be detected based on the label using, for example, a labeled secondary antibody.
- Cancer tissue contains cancer cells (ie, malignant tumor cells) and cancer stromal cells.
- MEFLIN is also expressed in malignant tumors (sarcomas) and benign tumors derived from mesenchymal stem cells or similar cells. Specifically, it is expressed on cancer cells in almost all cases of non-epithelial tumors such as osteosarcoma, chondrosarcoma, liposarcoma, rhabdomyosarcoma, undifferentiated polymorphic sarcoma, aggressive fibromatoma, and meningioma. Was confirmed. Therefore, the ADCs of the present invention can be used to treat malignant tumors (sarcomas) derived from mesenchymal stem cells or similar cells.
- the ADCs of the present invention can also be used to treat benign tumors.
- cancers cancers that develop from epithelial cells
- the MEFLIN protein is expressed in stromal cells (eg, CAF) in most cancers, even if the cancer cells themselves are MEFLIN protein negative. did. Therefore, by targeting the stromal cells of the carcinoma, the ADCs of the present invention can be used to treat the carcinoma.
- the expression of MEFLIN can be examined by, for example, in situ hybridization method or immunohistochemical staining using an antibody, but other methods such as RT-PCR method, Western blotting method, DNA microarray method, RNA sequence analysis method and the like can also be used. It is possible.
- the cancer to be treated with the ADC or the pharmaceutical composition of the present invention is not particularly limited, but cancer, for example, cancer originating from epithelial cells (cancer tumor), for example, lung cancer, pancreatic cancer, head and neck. Hmm, prostate cancer, bladder cancer, breast cancer, esophageal cancer, stomach cancer, colon cancer, uterine cancer, ovarian cancer, skin cancer, thyroid cancer, thoracic adenocarcinoma, kidney cancer, testicular cancer, Included are penile cancer, liver cancer, biliary tract cancer, biliary tract cancer, and these metastatic cancers (or cancer cells).
- cancer for example, cancer originating from epithelial cells (cancer tumor), for example, lung cancer, pancreatic cancer, head and neck. Hmm, prostate cancer, bladder cancer, breast cancer, esophageal cancer, stomach cancer, colon cancer, uterine cancer, ovarian cancer, skin cancer, thyroid cancer, thoracic adenocarcinoma, kidney cancer, testicular cancer, Include
- Cancers to be treated with the ADC or pharmaceutical composition of the present invention also include retroperitoneal tumors, vascular / lymphangiosarcoma, and metastatic cancers (or cancer cells) thereof. Cancers to be treated with the ADC or pharmaceutical composition of the present invention also include brain tumors and bone and soft tissue tumors. These cancers include cells whose cancer cells are MEFLIN protein positive or whose cancer stroma is MEFLIN protein positive even though the cancer cells are MEFLIN protein negative. According to the present invention, MEFLIN protein-positive cells are recruited into the cancer stroma. Therefore, according to the present invention, the cancer (or cancer cell) to be treated of the ADC or pharmaceutical composition of the present invention may be MEFLIN negative.
- MEFLIN protein-positive cells are recruited into the stromal of MEFLIN protein-negative cancer, and the ADC of the present invention targets the MEFLIN protein-positive cells recruited to the stromal and cancer (or cancer cells). ) Itself can exert antitumor activity against the cancer regardless of whether it is MEFLIN protein positive or negative. This can be explained by the bystander effect.
- the presence of MEFLIN protein-positive cells in the stroma of the cancer may be confirmed by examining the expression of MEFLIN protein in tissue donations.
- the cancer to be treated is MEFLIN protein negative, such as the presence of MEFLIN protein positive cells in the stroma of the cancer.
- the cancer to be treated of the ADC or pharmaceutical composition of the present invention is a MEFLIN protein-positive cancer (for example, sarcoma, for example, mucosal fibrosarcoma, malignant fibrous histiocytoma, liposarcoma, etc. It may be smooth myoma, rhabdomyosarcoma, malignant peripheral nerve sheath tumor, Ewing sarcoma, epithelial sarcoma, clear cell sarcoma, synovial sarcoma, and osteosarcoma).
- the cancer to be treated is MEFLIN protein positive and there are MEFLIN protein positive cells in the stroma of the cancer.
- the cancer to be treated is MEFLIN protein positive and no MEFLIN protein positive cells are detected in the stroma of the cancer.
- the cancer to be treated is a cancer that has been evaluated or presumed to be MEFLIN-positive, or the cancer cells themselves are MEFLIN-negative, but there are MEFLIN-positive cells in the stroma. It can be a cancer that has been evaluated or inferred.
- the bispecific antibody can be, for example, an antibody having a binding affinity for cancer cells and immune cells, respectively.
- the bispecific antibody can be, for example, an antibody that binds to surface antigens on T cells such as MEFLIN protein and CD3 protein.
- Bispecific antibodies can be IgG type, and diabody type. This embodiment may be effective, for example, for MEFLIN protein-positive cancer cells.
- the pharmaceutical composition comprises the ADC of the invention and an excipient.
- the pharmaceutical composition of the present invention can be administered by an administration method such as intravenous administration, subcutaneous administration, intratumoral administration, intraperitoneal administration, intraventricular administration, and intramuscular administration.
- the dose can be appropriately determined by the doctor in consideration of the patient's age, gender, body weight, severity of the disease, and the like.
- the use of the ADC of the present invention in the manufacture of a medicament for use in treating cancer is provided.
- the cancer to be treated can be a cancer (eg, carcinoma) that is MEFLIN protein negative and in which MEFLIN protein positive cells are present in the stroma of the cancer.
- the cancer can be a MEFLIN protein-positive cancer (eg, a sarcoma).
- a subject can be identified or selected by immunohistological staining with an anti-MEFLIN antibody in a tissue sample or tissue section thereof obtained from the subject or detection of MEFLIN mRNA by in situ hybridization.
- ADC of the present invention for use in a method of treating cancer is provided.
- ADCs of the present invention may be provided for use in methods of treating cancer.
- Example 1 Preparation of antibody and analysis of the characteristics of the obtained monoclonal antibody
- a monoclonal antibody that recognizes the human MEFLIN protein was prepared, and the binding characteristics of the antibody were confirmed by various assays.
- Mouse Meflin stable expression human fetal kidney (HEK293) cell line (hereinafter: mMeflin HEK293 cell line) and mouse Meflin stable expression Chinese hamster ovary (CHO) cell line (hereinafter “mMeflin CHO cell line”) by Flp-In TM system (Thermo Fisher) ") was established.
- mMeflin CHO cell line mouse Meflin stable expression Chinese hamster ovary (CHO) cell line
- hMeflin HEK293 cell line mouse Meflin stable expression Chinese hamster ovary (CHO) cell line
- hMEFLIN CHO cell line human MEFLIN stable expression Chinese hamster ovary
- Human MEFLIN recombinant protein was used as an antigen for animal immunization for producing human MEFLIN antibody.
- Human MEFLIN recombinant protein was obtained by a mammalian cell expression system.
- the human MEFLIN gene was introduced into a human fetal kidney-derived cell line (Expi293F cells, Thermo Fisher) acclimatized to suspension culture by the lipofection method (lipofectamine2000, invitrogen).
- lipofection method lipofectamine2000, invitrogen.
- the GPI anchor sequence at the carboxyl terminus of the MEFLIN gene was removed to allow subsequent purification, and a histidine tag was introduced therein.
- the cell culture medium 5 to 8 days after gene transfer is collected, filtered (0.22 ⁇ m), and then subjected to a column packed with a carrier (Ni Excel, GE Healthcare) that specifically binds a histidine-tagged protein.
- the MEFLIN protein in the culture medium is bound by.
- MEFLIN protein bound by buffer solution containing imidazole (500 mM) was competitively eluted.
- the final human MEFLIN recombinant protein was obtained by removing imidazole from the eluate containing the MEFLIN protein by ultrafiltration (dialysis using phosphate buffer).
- the purity of the purified human MEFLIN recombinant protein was measured by subjecting the protein to Coomassie Brilliant Blue staining after SDS-PAGE electrophoresis.
- the soles of the hind limbs of WKY rats were immunized once with 100 ⁇ g of recombinant human MEFLIN protein (purification degree 95%).
- plasma cells present in the iliac lymph nodes and a myeloma (SP2 / 0) cell line were electrically fused to prepare a hybridoma.
- HAT medium (MPB) and BM-condimed H1 (Roche) were added to High glucose DMEM (Nacalai), and hybridomas were screened.
- a monoclonal antibody was prepared in the same manner as in the preparation of a rat anti-human MEFLIN monoclonal antibody, except that 50 ⁇ g of recombinant human MEFLIN protein (purification degree 95%) was immunized once on the C57BL / 6 mouse ridge muscle.
- a pRP-CMV vector in which a DNA encoding a human MEFLIN protein or a fragment thereof was inserted into a human fetal kidney (HEK293) cell line was transiently expressed by a lipofection method to prepare a cytolytic solution.
- the fragment is the fragment shown in FIG. 1 or 2, and has a G196 tag and a His tag at the C-terminal.
- SDS-PAGE was performed using the cytolytic solution, and the electrophoresed protein was transferred to the PVDF membrane.
- the PVDF membrane was reacted with 5% skim milk / PBS at room temperature for 1 hour.
- As the primary antibody a culture supernatant of each hybridoma clone diluted 50-fold with PBS was used.
- an anti-G196 antibody was used as the primary antibody. It was reacted with the PVDF membrane at 4 ° C. overnight in the presence of the primary antibody. The PVDF membrane was then washed with 0.05% Tween / PBS.
- an anti-rat or anti-mouse antibody-horseradish peroxidase (HRP) conjugate was used as the secondary antibody. The secondary antibody was diluted 1000-fold with PBS and reacted with the PVDF membrane at room temperature for 45 minutes. After washing with 0.05% Tween / PBS, the HRP labeling the antibody was made to emit light with ECL (GE Healthcare), and the image was captured in a computer with a CCD imager (Las4000, GE Healthcare).
- the hMeflin CHO cell line or mMeflin CHO cell line was cultured at 37 ° C. and 5% CO 2 for 3 hours in an assay solution containing 20 mM HEPS and 0.1% BSA in F-12 Glutamax culture medium (Gibco). Each hybridoma supernatant was diluted 10-fold with the assay solution, added to the above CHO cell line, and cultured for 15 minutes.
- the CHO cell line was washed with PBS (4 ° C.) and washed with a washing solution (MilliQ-H 2 O containing 0.5 M NaCl, 3% acetic acid) to remove the antibody bound on the cell membrane.
- the CHO cell line was reacted with 4% paraformaldehyde (PFA) for 10 minutes to fix it, and then reacted with 0.1% TiritonX-100 for 5 minutes to perform cell membrane permeabilization treatment.
- PFA paraformaldehyde
- As the secondary antibody a secondary antibody using an anti-rat or anti-mouse antibody-Alexa488 conjugate was diluted 400-fold with PBS and reacted with cells subjected to cell membrane permeabilization for 30 minutes at room temperature. After washing with PBS, it was reacted with DAPI at room temperature for 5 minutes, washed with PBS, and images were acquired with a confocal microscope (LSM700, ZEISS).
- ISH staining in situ hybridization method
- ISLR-positive tumor tissues those in which 20% or more of the cells having a fibroblast-like morphology infiltrating the cancer stroma are ISLR-positive are designated as ISLR-positive tumor tissues, and those in less than that are negative tumor tissues. Grouped as. If a signal was observed in at least a part of the cytoplasm, it was considered to be ISLR positive.
- an anti-MEFLIN monoclonal antibody and an anti-catepsin K antibody were used as the primary antibody, and an anti-rat antibody-Alexa488 conjugate and an anti-mouse antibody-Alexa594 conjugate were used as the secondary antibody.
- Invitrogen was used as an immunofluorescent double staining method. The intracellular nuclei were stained with DAPI. MEFLIN (green), cathepsin K (red), nucleus (blue). Images were acquired with a confocal microscope (LSM700, ZEISS).
- the region with the highest tumor component is selected by microscopic examination from the histopathological specimens obtained from cancer patients, and the expression of MEFLIN (also referred to as "ISLR") is immunohistochemical staining (hereinafter referred to as "IHC staining"). I looked it up in. Among the examined tissues, the tissues were observed with a medium-magnification visual field (20x objective lens), and 20% or more of the cells having a fibroblast-like morphology infiltrating the tumor stroma were ISLR-positive. Tumor tissue was defined, and less was grouped as negative tumor tissue. If a signal was observed in at least a part of the cytoplasm, it was considered to be ISLR positive. An anti-rat or anti-mouse antibody-horseradish peroxidase (HRP) conjugate was used as the secondary antibody (ImmPRESS, VECTORS LABORATORIES).
- HRP anti-rat or anti-mouse antibody-horseradish peroxidase
- FIG. 1 The results of confirming the binding of each monoclonal antibody to each human MEFLIN protein fragment by Western blotting were as shown in FIG.
- a rat or mouse-derived monoclonal antibody produced from clone 27-7 (hereinafter, simply referred to as “27-7 antibody”) is bound to the amino acid regions 1 to 343 of human MEFLIN. It was bound to the 1st to 399th amino acid regions. This suggests that the 27-7 antibody binds to the amino acid region 344 to 399 of human MEFLIN.
- FIG. 1 suggests that the 34-4 antibody binds to the 232-343th amino acid region; the 35-9 antibody is suggested to be the 1st to 146th amino acid region; the 27-8 antibody.
- the 3-2 antibody, the 11-8 antibody, the 12-2 antibody, the 19-7 antibody, and the 23-1 antibody bind to the amino acid regions 344 to 399 of human MEFLIN. It is suggested that the 13-1 antibody binds to the 147-231 amino acid region; the 16-5 antibody, 22-3 antibody, and 32-1 antibody bind to the 1-146th amino acid region. It was suggested to do.
- FIGS. 3-5 The internalizing activity of each antibody clone was confirmed. The results were as shown in FIGS. 3-5. As shown in FIGS. 3-5, 21-3 antibody, 25-1 antibody, 8-2 antibody, 27-7 antibody, 27-8 antibody, 34-4 antibody, 35-9 antibody, 46-3 antibody, With 3-2 antibody, 11-8 antibody, 12-2 antibody, 16-5 antibody, 19-7 antibody, 22-3 antibody, 23-1 antibody and 32-1 antibody, a strong signal derived from the antibody is emitted inside the cell. Observed.
- MEFLIN protein in various cancer tissues was as shown in Table 10.
- Table 10 shows the number of confirmed tissue samples, the number of samples that were positive, and the positive rate (%).
- the expression of human MEFLIN (ISLR) in the tumor stroma was as shown in Table 11.
- Table 11 shows the number of confirmed tissue samples, the number of samples that were positive, and the positive rate (%). In addition, 2 out of 7 cases of gastric cancer were positive.
- human MEFLIN was negative in tumor cells of pancreatic cancer, lung cancer, breast cancer, colon cancer, gastric cancer, bile duct cancer, ovarian cancer, bladder cancer and esophageal cancer.
- Example 2 Preparation of antibody-drug conjugate (ADC) and its in vivo drug efficacy test
- ADC antibody-drug conjugate
- a conjugate of the antibody prepared in Example 1 and a cytotoxic agent was prepared.
- the prepared conjugate was administered to a cancer-bearing mouse model, and the antitumor effect was confirmed.
- Antibodies were purified from the antibody-containing hybridoma supernatant concentrated by high-density culture of hybridomas using Mono Spin TM L ProG (GL Science). The disulfide bond of the antibody was cleaved by performing a reduction treatment with 1.0 mM DTT at 25 ° C. for 30 minutes, and VcMMAE (mc-vc-PAB-MMAE, MedChemExpress) or mc-PEG 12-vc-PABC was applied to the exposed thiol group. -MMAE was added at room temperature and purified by limit filtration. VcMMAE is a compound registered under CAS Registry Number 646502-53-6.
- VcMMAE monomethyl auristatin E (MMAE) is bound to valine-citrulline linker via p-aminobenzoyloxycarbonyl (PBAC), and when valine-citrulline is cleaved by a protease such as catepsin B, MMAE is released. It is a mechanism to be released.
- the valine-citrulline linker is modified with a maleimide caproyl group and can react with the thiol group of the antibody cysteine via the maleimide group.
- mc-PEG 12- vc-PABC-MMAE is similar to VcMMAE except that it is mediated by PEG, and the mechanism of MMAE release is also the same as VcMMAE.
- the mc-vc-PAB-MMAE has the structure of the following formula (I).
- the resulting ADC may have the structure of formula (II) below.
- n is a natural number from 1 to 8.
- the mc-PEG 12- vc-PABC-MMAE has the structure of the following formula (III).
- Cytotoxicity of ADC to human MEFLIN-expressing HEK293 cells was tested. Specifically, the hMeflin HEK293 cell line was seeded on a plate at 1.0 ⁇ 10 4 / well and cultured at 37 ° C. and 5% CO 2 for 1 day. Next, ADCs of each concentration were added, and the cells were cultured for 2 days, and the cell proliferation ability was evaluated by the MTT test. Cell proliferation ability was evaluated by measuring the absorbance of the sample at a wavelength of 590 nm with an absorptiometer (POWERSCAN4, DS PHARMA BIOMEDICAL) in the MMT test.
- an absorptiometer (POWERSCAN4, DS PHARMA BIOMEDICAL)
- Meflin in the hMeflin HEK293 cell line was evaluated by FACS analysis.
- clone number when the clone number is simply described in the figure, it means that the monoclonal antibody itself was used, and when the clone number is described immediately before or after the ADC, the antibody is the clone.
- CTL ADC or isotype control IgG ADC means that the ADC used is an isotype control IgG antibody.
- Rat IgG2a BioLegend was used as an isotype control antibody.
- ADC Cytotoxicity Against Human Rhabdomyosarcoma Cell Lines Similar to the above, ADC cytotoxicity against rhabdomyosarcoma (KYM-1) cell lines was tested. The results were as shown in FIG. As shown in FIG. 7, it was clear from FACS analysis that the rhabdomyosarcoma cell line strongly expressed the human MEFLIN protein. In addition, ADC27-7 and ADC34-4 significantly reduced the proliferation of the KYM-1 cell line.
- the cytotoxicity of ADC in a subcutaneous transplant model of a rhabdomyosarcoma cell line was tested using a cancer-bearing mouse model.
- ADC was intraperitoneally administered to mice having a tumor volume of about 100 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of tumors formed by transplantation of KYM cell lines was performed.
- HsOs1 Meflin-positive osteosarcoma
- ADC cytotoxic endogenous Meflin-negative human pancreatic cancer (BxPC-3) cell line 1.0 ⁇ 10 7 cells of female nude mice (BALB / cSlc nu / nu)
- BxPC-3 cell line 1.0 ⁇ 10 7 cells of female nude mice (BALB / cSlc nu / nu)
- ADC was intraperitoneally administered to mice having a tumor volume of about 150 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of the tumor formed by transplantation of the BxPC-3 cell line was performed.
- pancreatic cancer cells themselves were MEFLIN-negative, but mouse MEFLIN-positive cells were observed in the tumor stromal cells. It was also revealed that 21-3ADC and 25-1ADC have strong antitumor activity against pancreatic cancer. Since the original tissue was MEFLIN-negative, it was revealed that MEFLIN-positive cells were recruited to the vicinity of the tumor. It was also suggested that ADC exerts antitumor activity against tumors.
- Neuroblastoma ADC in a subcutaneous transplant model of cell lines Cytotoxicity addition, were implanted subcutaneously into endogenous Meflin positive neuroblastoma (NB-1) cell lines 1.0 ⁇ 10 7 pieces of the back of female NOD SCID mice ( Each group n 5).
- DLD-1 cytotoxic endogenous Meflin-negative human colorectal cancer
- ISH staining of tumors formed by transplantation of MKN45 cell line was performed.
- FIG. 15 analyzed single-cell RNA sequence data (Tabula Muris) (see Non-Patent Document 7) of a cell population derived from mouse pancreas published on the Internet.
- tSNE t-distributed Stochastic Neighbor Embedding; t-distributed stochastic neighborhood embedding
- multidimensional data composed of a large amount of RNA expression level data is compressed to one dimension by nonlinear conversion. Represents the expressed numerical value. That is, in FIG.
- cells can be clustered based on the gene expression profile by developing a large amount of RNA expression level data expressed in individual cells in a two-dimensional plane by tSNE1 and tSNE2.
- tSNE1 and tSNE2 RNA expression level data expressed in individual cells in a two-dimensional plane.
- tSNE1 and tSNE2 it is possible to understand what kind of cell cluster the individual cells indicated by the circles form. This makes it possible to confirm whether or not the cell group expressing a specific factor matches the cell group expressing another specific factor in the clustering method, and in addition, which cell the cell group is expressed in. It is also possible to decide whether to correspond to the species.
- Pancreatic stellate cells in the mouse pancreas express both MEFLIN and cathepsin K, and that the MEFLIN-positive cell population and the cathepsin K-positive cell population match in the mouse pancreas. (See the arrow in the figure).
- FIG. 16 analyzed single-cell RNA sequence data (Tabula Muris) (see Non-Patent Document 7) of a cell population derived from mouse lung published on the Internet. It is shown that both MEFLIN and cathepsin K are expressed in stromal cells of mouse lungs, indicating that the MEFLIN-positive cell population and the cathepsin K-positive cell population match in the mouse lung (in the figure). See arrow). (See the arrow in the figure).
- the full length of the mouse MEFLIN protein was expressed in CHO cells, and Western blotting was attempted on the cytolytic solution. The results were as shown in FIG. As shown in FIG. 17, expression of the full length of the mouse MEFLIN protein in CHO cells increased the expression of endogenous cathepsin K.
- FIG. 18 shows immunofluorescence using an anti-MEFLIN monoclonal antibody and an anti-catepsin K antibody against the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with an osteosarcoma cell line (HsOs1). The result of performing the double staining method is shown. It indicates that cathepsin K (red) is secreted around MEFLIN-positive cells (green) (arrow).
- FIG. 19 shows anti-MEFLIN monoclonal antibody and anti-catepsin K antibody against the tumor tissue of a cancer-bearing model mouse subcutaneously transplanted with a pancreatic cancer cell line (BxPC-3). The result of performing the immunofluorescent double staining method used is shown. It indicates that cathepsin K (red) is secreted around MEFLIN-positive cells (green) (arrow).
- MEFLIN-negative cancer cells when human MEFLIN-negative cancer cells are transplanted into mice, MEFLIN-positive cells are recruited to tissues containing the cancer cells, and the recruited MEFLIN-positive cells are targeted.
- the ADC has been shown to be beneficial in treating cancer. It has also been shown that ADCs that target the human MEFLIN protein do not necessarily require internalizing activity. Further, in the stroma containing MEFLIN protein-positive cells, an enzyme such as cathepsin that cleaves the valine-citrulline linker is present, and it is considered that the ADC that is not taken up into the cells exhibits cytotoxicity.
- ADC was administered to the tail vein to mice having a tumor volume of about 250 mm 3 (5 mg / kg body weight, 3 times in total at 4-day intervals).
- ISH staining of tumors formed by transplantation of HuCCT1 cell line was performed.
- the results were as shown in FIG. As shown in the histological image of FIG. 20, the cholangiocarcinoma cells themselves were MEFLIN-negative, but mouse MEFLIN-positive cells were observed in the tumor stromal cells. It was also revealed that 21-3ADC and 25-1ADC have strong antitumor activity against bile duct cancer. Since the original tissue was MEFLIN-negative, it was revealed that MEFLIN-positive cells were recruited to the vicinity of the tumor. It was also suggested that ADC exerts antitumor activity against tumors.
- ADC was administered to the tail vein to mice having a tumor volume of about 300 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of the tumor formed by transplantation of the T24 cell line was performed.
- the bladder cancer cells themselves were MEFLIN-negative, but mouse MEFLIN-positive cells were observed in the tumor stromal cells. It was also revealed that 21-3ADC and 25-1ADC have weak antitumor activity against bladder cancer. Since the original tissue was MEFLIN-negative, it was revealed that MEFLIN-positive cells were recruited to the vicinity of the tumor. It was also suggested that ADC exerts antitumor activity against tumors.
- ADC was intraperitoneally administered to mice having a tumor volume of about 100 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of the tumor formed by transplantation of the OV-90 cell line was performed.
- the results were as shown in FIG. As shown in the histological image of FIG. 22, the ovarian cancer cells themselves were MEFLIN-negative, but mouse MEFLIN-positive cells were observed in the tumor stromal cells. It was also revealed that 21-3ADC and 25-1ADC have strong antitumor activity against ovarian cancer. Since the original tissue was MEFLIN-negative, it was revealed that MEFLIN-positive cells were recruited to the vicinity of the tumor. It was also suggested that ADC exerts antitumor activity against tumors.
- ADC was intraperitoneally administered to mice having a tumor volume of about 200 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of tumors formed by transplantation of KYSE cell lines was performed.
- HsOs1 cytotoxic endogenous Meflin-positive osteosarcoma
- ADC was administered to the tail vein to mice having a tumor volume of about 250 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of tumors formed by transplantation of HsOs1 cell line was performed.
- ADC was administered to the tail vein to mice having a tumor volume of about 300 mm 3 (5 mg / kg body weight, 5 times in total at 4-day intervals).
- ISH staining of tumors formed by transplantation of MCF-7 cell lines was performed.
- the results were as shown in FIG. As shown in the histological image of FIG. 25, the breast cancer cells themselves were MEFLIN-negative, but mouse MEFLIN-positive cells were observed in the tumor stromal cells. It was also revealed that 21-3ADC and 25-1ADC have weak antitumor activity against breast cancer. Since the original tissue was MEFLIN-negative, it was revealed that MEFLIN-positive cells were recruited to the vicinity of the tumor. It was also suggested that ADC exerts antitumor activity against tumors.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Immunology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Endocrinology (AREA)
- Reproductive Health (AREA)
- Pulmonology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
[1]MEFLINに結合する抗体と細胞傷害剤との抗体-薬物コンジュゲート(ADC)を含む、がんを処置することに用いるための医薬組成物。
[2]前記抗体は、インターナライズ活性を有する、上記[1]に記載の医薬組成物。
[3]ADCが、抗体と薬物とがリンカーを介して連結されたADCであり、リンカーが、細胞内で開裂される切断部位を有する、上記[1]または[2]に記載の医薬組成物。
[4]MEFLINに結合する抗体であって、以下からなる群から選択される抗体:
(1A)配列番号1に記載のアミノ酸配列を有する重鎖CDR1、配列番号2に記載のアミノ酸配列を有する重鎖CDR2、および配列番号3に記載のアミノ酸配列を有する重鎖CDR3を含む重鎖可変領域と、配列番号4に記載のアミノ酸配列を有する軽鎖CDR1、配列番号5に記載のアミノ酸配列を有する軽鎖CDR2、および配列番号6に記載のアミノ酸配列を有する軽鎖CDR3を含む軽鎖可変領域とを有する抗体;
(1B)配列番号7に記載のアミノ酸配列を有する重鎖可変領域と、配列番号8に記載のアミノ酸配列を有する軽鎖可変領域とを有する抗体;
(1C)上記(1B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(1D)上記(1B)の抗体とMEFLINタンパク質上で重なるエピトープに結合する抗体;
(2A)配列番号9に記載の重鎖CDR1と、配列番号10に記載の重鎖CDR2と、配列番号11に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号12に記載の軽鎖CDR1と、配列番号13に記載の軽鎖CDR2と、配列番号14に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(2B)配列番号15に記載の重鎖可変領域と、配列番号16に記載の軽鎖可変領域を有する抗体;
(2C)上記(2B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(2D)上記(2B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(3A)配列番号17に記載の重鎖CDR1と、配列番号18に記載の重鎖CDR2と、配列番号19に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号20に記載の軽鎖CDR1と、配列番号21に記載の軽鎖CDR2と、配列番号22に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(3B)配列番号23に記載の重鎖可変領域と、配列番号24に記載の軽鎖可変領域を有する抗体;
(3C)上記(3B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(3D)上記(3B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(4A)配列番号25に記載の重鎖CDR1と、配列番号26に記載の重鎖CDR2と、配列番号27に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号28に記載の軽鎖CDR1と、配列番号29に記載の軽鎖CDR2と、配列番号30に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(4B)配列番号31に記載の重鎖可変領域と、配列番号32に記載の軽鎖可変領域を有する抗体;
(4C)上記(4B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(4D)上記(4B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(5A)配列番号33に記載の重鎖CDR1と、配列番号34に記載の重鎖CDR2と、配列番号35に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号36に記載の軽鎖CDR1と、配列番号37に記載の軽鎖CDR2と、配列番号38に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(5B)配列番号39に記載の重鎖可変領域と、配列番号40に記載の軽鎖可変領域を有する抗体;
(5C)上記(5B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(5D)上記(5B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(6A)配列番号41に記載の重鎖CDR1と、配列番号42に記載の重鎖CDR2と、配列番号43に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号44に記載の軽鎖CDR1と、配列番号45に記載の軽鎖CDR2と、配列番号46に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(6B)配列番号47に記載の重鎖可変領域と、配列番号48に記載の軽鎖可変領域を有する抗体;
(6C)上記(6B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(6D)上記(6B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(7A)配列番号49に記載の重鎖CDR1と、配列番号50に記載の重鎖CDR2と、配列番号51に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号52に記載の軽鎖CDR1と、配列番号53に記載の軽鎖CDR2と、配列番号54に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(7B)配列番号55に記載の重鎖可変領域と、配列番号56に記載の軽鎖可変領域を有する抗体;
(7C)上記(7B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(7D)上記(7B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(8A)配列番号57に記載の重鎖CDR1と、配列番号58に記載の重鎖CDR2と、配列番号59に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号60に記載の軽鎖CDR1と、配列番号61に記載の軽鎖CDR2と、配列番号62に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(8B)配列番号63に記載の重鎖可変領域と、配列番号64に記載の軽鎖可変領域を有する抗体;
(8C)上記(8B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(8D)上記(8B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(9A)配列番号65に記載の重鎖CDR1と、配列番号66に記載の重鎖CDR2と、配列番号67に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号68に記載の軽鎖CDR1と、配列番号69に記載の軽鎖CDR2と、配列番号70に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(9B)配列番号71に記載の重鎖可変領域と、配列番号72に記載の軽鎖可変領域を有する抗体;
(9C)上記(9B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;
および、
(9D)上記(9B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(10A)配列番号73に記載の重鎖CDR1と、配列番号74に記載の重鎖CDR2と、配列番号75に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号76に記載の軽鎖CDR1と、配列番号77に記載の軽鎖CDR2と、配列番号78に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(10B)配列番号79に記載の重鎖可変領域と、配列番号80に記載の軽鎖可変領域を有する抗体;
(10C)上記(10B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(10D)上記(10B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(11A)配列番号81に記載の重鎖CDR1と、配列番号82に記載の重鎖CDR2と、配列番号83に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号84に記載の軽鎖CDR1と、配列番号85に記載の軽鎖CDR2と、配列番号86に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(11B)配列番号87に記載の重鎖可変領域と、配列番号88に記載の軽鎖可変領域を有する抗体;
(11C)上記(11B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(11D)上記(11B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(12A)配列番号89に記載の重鎖CDR1と、配列番号90に記載の重鎖CDR2と、配列番号91に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号92に記載の軽鎖CDR1と、配列番号93に記載の軽鎖CDR2と、配列番号94に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(12B)配列番号95に記載の重鎖可変領域と、配列番号96に記載の軽鎖可変領域を有する抗体;
(12C)上記(12B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(12D)上記(12B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(13A)配列番号97に記載の重鎖CDR1と、配列番号98に記載の重鎖CDR2と、配列番号99に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号100に記載の軽鎖CDR1と、配列番号101に記載の軽鎖CDR2と、配列番号102に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(13B)配列番号103に記載の重鎖可変領域と、配列番号104に記載の軽鎖可変領域を有する抗体;
(13C)上記(13B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(13D)上記(13B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(14A)配列番号105に記載の重鎖CDR1と、配列番号106に記載の重鎖CDR2と、配列番号107に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号108に記載の軽鎖CDR1と、配列番号109に記載の軽鎖CDR2と、配
列番号110に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(14B)配列番号111に記載の重鎖可変領域と、配列番号112に記載の軽鎖可変領域を有する抗体;
(14C)上記(14B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(14D)上記(14B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(15A)配列番号113に記載の重鎖CDR1と、配列番号114に記載の重鎖CDR2と、配列番号115に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号116に記載の軽鎖CDR1と、配列番号117に記載の軽鎖CDR2と、配列番号118に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(15B)配列番号119に記載の重鎖可変領域と、配列番号120に記載の軽鎖可変領域を有する抗体;
(15C)上記(15B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(15D)上記(15B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(16A)配列番号121に記載の重鎖CDR1と、配列番号122に記載の重鎖CDR2と、配列番号123記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号124に記載の軽鎖CDR1と、配列番号125に記載の軽鎖CDR2と、配列番号126に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(16B)配列番号127に記載の重鎖可変領域と、配列番号128に記載の軽鎖可変領域を有する抗体;
(16C)上記(16B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(16D)上記(16B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;並びに
(17A)配列番号129に記載の重鎖CDR1と、配列番号130に記載の重鎖CDR2と、配列番号131記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号132に記載の軽鎖CDR1と、配列番号133に記載の軽鎖CDR2と、配列番号134に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(17B)配列番号135に記載の重鎖可変領域と、配列番号136に記載の軽鎖可変領域を有する抗体;
(17C)上記(17B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(17D)上記(17B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体。
[5]上記[4]に記載の抗体と、細胞傷害剤との抗体-薬物コンジュゲート(ADC)を含む、医薬組成物。
[6]がんを処置することに用いるための、上記[5]に記載の医薬組成物。
[7]がんが、肉腫である、上記[1]~[3]および[6]のいずれかに記載の医薬組成物。
[8]がんが、MEFLIN陽性である肉腫である、上記[7]に記載の医薬組成物。
[9]肉腫が、粘膜線維肉腫、悪性線維性組織球腫、脂肪肉腫、平滑筋肉腫、横紋筋肉腫、神経芽腫、悪性末梢神経鞘腫瘍、ユーイング肉腫、類上皮肉腫、明細胞肉腫、滑膜肉腫、および骨肉腫からなる群から選択される肉腫である、上記[7]または[8]に記載の医薬組成物。
[10]がんが、癌腫である、上記[1]~[3]および[6]のいずれかに記載の医薬組成物。
[11]がんが、乳がん、膵臓がん、肺がん、大腸がん、胃がん、胆管がん、卵巣がん、膀胱がん、および食道がんからなる群から選択されるがんである、上記[10]に記載の医薬組成物。
[12]がんが、MEFLIN陰性であり、当該がん周囲の間質がMEFLIN陽性の細胞を含む、上記[1]~[3]、[6]、[10]および[11]のいずれかに記載の医薬組成物。
[13]抗体が、インターナライズ活性を有しない、上記[10]~[12]のいずれかに記載の医薬組成物。
[14]抗体が、インターナライズ活性を有する、上記[7]~[9]のいずれかに記載の医薬組成物。
[15]ADCが、抗体と薬物とがリンカーを介して連結されたADCであり、リンカーが、開裂性リンカーである、上記[13]に記載の医薬組成物。
[16]リンカーが、カテプシンKにより開裂する開裂性リンカーである、上記[15]に記載の医薬組成物。
[17]リンカーが、バリン-シトルリンのジペプヂドを含み、カテプシンK存在下で切断される、上記[16]に記載の医薬組成物。
[18]リンカーが、非開裂性リンカーである、上記のいずれかに記載の医薬組成物。
ある態様では、リンカーは、第1のスペーサーおよび開裂性部分を含む。ある態様では、リンカーは、第1のスペーサー、開裂性部分、および第2のスペーサーを含む。ある特定の対象では、リンカーは、PEG、開裂性部分、およびPABCを含む。
抗体とリンカーとの結合は、例えば抗体のスルフヒドリル基にマレイミド基を介して連結することができる。
ある態様では、抗体は、そのスルフヒドリル基を介して、マレイミド-PEG-開裂性部分を有するリンカーで抗がん剤と連結されている。ある態様では、抗体は、そのスルフヒドリル基を介して、マレイミド-PEG-開裂性部分-PABCを有するリンカーで抗がん剤と連結されている。
ある態様では、ADCは、以下の式(II)に示される構造を有し得る。
本発明のADCはがんの治療薬として有用であると考えられる。
(1A)配列番号1に記載の重鎖CDR1と、配列番号2に記載の重鎖CDR2と、配列番号3に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号4に記載の軽鎖CDR1と、配列番号5に記載の軽鎖CDR2と、配列番号4に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(1B)配列番号7に記載の重鎖可変領域と、配列番号8に記載の軽鎖可変領域を有する抗体;
(1C)上記(1B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(1D)上記(1B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(2A)配列番号9に記載の重鎖CDR1と、配列番号10に記載の重鎖CDR2と、配列番号11に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号12に記載の軽鎖CDR1と、配列番号13に記載の軽鎖CDR2と、配列番号14に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(2B)配列番号15に記載の重鎖可変領域と、配列番号16に記載の軽鎖可変領域を有する抗体;
(2C)上記(2B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(2D)上記(2B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(3A)配列番号17に記載の重鎖CDR1と、配列番号18に記載の重鎖CDR2と、配列番号19に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号20に記載の軽鎖CDR1と、配列番号21に記載の軽鎖CDR2と、配列番号22に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(3B)配列番号23に記載の重鎖可変領域と、配列番号24に記載の軽鎖可変領域を有する抗体;
(3C)上記(3B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(3D)上記(3B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(4A)配列番号25に記載の重鎖CDR1と、配列番号26に記載の重鎖CDR2と、配列番号27に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号28に記載の軽鎖CDR1と、配列番号29に記載の軽鎖CDR2と、配列番号30に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(4B)配列番号31に記載の重鎖可変領域と、配列番号32に記載の軽鎖可変領域を有する抗体;
(4C)上記(4B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(4D)上記(4B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(5A)配列番号33に記載の重鎖CDR1と、配列番号34に記載の重鎖CDR2と、
配列番号35に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号36に記載の軽鎖CDR1と、配列番号37に記載の軽鎖CDR2と、配列番号38に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(5B)配列番号39に記載の重鎖可変領域と、配列番号40に記載の軽鎖可変領域を有する抗体;
(5C)上記(5B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(5D)上記(5B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(6A)配列番号41に記載の重鎖CDR1と、配列番号42に記載の重鎖CDR2と、配列番号43に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号44に記載の軽鎖CDR1と、配列番号45に記載の軽鎖CDR2と、配列番号46に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(6B)配列番号47に記載の重鎖可変領域と、配列番号48に記載の軽鎖可変領域を有する抗体;
(6C)上記(6B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(6D)上記(6B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(7A)配列番号49に記載の重鎖CDR1と、配列番号50に記載の重鎖CDR2と、配列番号51に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号52に記載の軽鎖CDR1と、配列番号53に記載の軽鎖CDR2と、配列番号54に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(7B)配列番号55に記載の重鎖可変領域と、配列番号56に記載の軽鎖可変領域を有する抗体;
(7C)上記(7B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(7D)上記(7B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(8A)配列番号57に記載の重鎖CDR1と、配列番号58に記載の重鎖CDR2と、配列番号59に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号60に記載の軽鎖CDR1と、配列番号61に記載の軽鎖CDR2と、配列番号62に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(8B)配列番号63に記載の重鎖可変領域と、配列番号64に記載の軽鎖可変領域を有する抗体;
(8C)上記(8B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(8D)上記(8B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(9A)配列番号65に記載の重鎖CDR1と、配列番号66に記載の重鎖CDR2と、配列番号67に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号68に記載の軽鎖CDR1と、配列番号69に記載の軽鎖CDR2と、配列番号70に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(9B)配列番号71に記載の重鎖可変領域と、配列番号72に記載の軽鎖可変領域を有する抗体;
(9C)上記(9B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(9D)上記(9B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(10A)配列番号73に記載の重鎖CDR1と、配列番号74に記載の重鎖CDR2と、配列番号75に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号76に記載の軽鎖CDR1と、配列番号77に記載の軽鎖CDR2と、配列番号78に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(10B)配列番号79に記載の重鎖可変領域と、配列番号80に記載の軽鎖可変領域を有する抗体;
(10C)上記(10B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(10D)上記(10B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(11A)配列番号81に記載の重鎖CDR1と、配列番号82に記載の重鎖CDR2と、配列番号83に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号84に記載の軽鎖CDR1と、配列番号85に記載の軽鎖CDR2と、配列番号86に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(11B)配列番号87に記載の重鎖可変領域と、配列番号88に記載の軽鎖可変領域を有する抗体;
(11C)上記(11B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(11D)上記(11B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(12A)配列番号89に記載の重鎖CDR1と、配列番号90に記載の重鎖CDR2と、配列番号91に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号92に記載の軽鎖CDR1と、配列番号93に記載の軽鎖CDR2と、配列番号94に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(12B)配列番号95に記載の重鎖可変領域と、配列番号96に記載の軽鎖可変領域を有する抗体;
(12C)上記(12B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(12D)上記(12B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(13A)配列番号97に記載の重鎖CDR1と、配列番号98に記載の重鎖CDR2と、配列番号99に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号100に記載の軽鎖CDR1と、配列番号101に記載の軽鎖CDR2と、配列番号102に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(13B)配列番号103に記載の重鎖可変領域と、配列番号104に記載の軽鎖可変領域を有する抗体;
(13C)上記(13B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(13D)上記(13B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(14A)配列番号105に記載の重鎖CDR1と、配列番号106に記載の重鎖CDR2と、配列番号107に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号108に記載の軽鎖CDR1と、配列番号109に記載の軽鎖CDR2と、配列番号110に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(14B)配列番号111に記載の重鎖可変領域と、配列番号112に記載の軽鎖可変領域を有する抗体;
(14C)上記(14B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(14D)上記(14B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(15A)配列番号113に記載の重鎖CDR1と、配列番号114に記載の重鎖CDR2と、配列番号115に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号116に記載の軽鎖CDR1と、配列番号117に記載の軽鎖CDR2と、配列番号118に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(15B)配列番号119に記載の重鎖可変領域と、配列番号120に記載の軽鎖可変領域を有する抗体;
(15C)上記(15B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(15D)上記(15B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(16A)配列番号121に記載の重鎖CDR1と、配列番号122に記載の重鎖CDR2と、配列番号123記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号124に記載の軽鎖CDR1と、配列番号125に記載の軽鎖CDR2と、配列番号126に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(16B)配列番号127に記載の重鎖可変領域と、配列番号128に記載の軽鎖可変領域を有する抗体;
(16C)上記(16B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(16D)上記(16B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
(17A)配列番号129に記載の重鎖CDR1と、配列番号130に記載の重鎖CDR2と、配列番号131記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号132に記載の軽鎖CDR1と、配列番号133に記載の軽鎖CDR2と、配列番号134に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(17B)配列番号135に記載の重鎖可変領域と、配列番号136に記載の軽鎖可変領域を有する抗体;
(17C)上記(17B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(17D)上記(17B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体
が提供される。当該抗体は、好ましくは、ヒトキメラ抗体であり、好ましくは、ヒト化抗体であり得る。
本実施例では、ヒトMEFLINタンパク質を認識するモノクローナル抗体を調製し、当該抗体の結合特性を様々なアッセイによって確認した。
Flp-InTMシステム(Thermo Fisher)によってマウスMeflin安定発現ヒト胎児腎(HEK293)細胞株(以下:mMeflin HEK293細胞株)とマウスMeflin安定発現チャイニーズハムスター卵巣(CHO)細胞株(以下「mMeflin CHO細胞株」という)を樹立した。同様に、ヒトMEFLIN安定発現ヒト胎児腎(HEK293)細胞株(以下:hMeflin HEK293細胞株)とヒトMEFLIN安定発現チャイニーズハムスター卵巣(CHO)細胞株(以下「hMEFLIN CHO細胞株」という)を樹立した。
ヒトMEFLIN抗体を作製するための動物免疫にはヒトMEFLIN組換えタンパク質を抗原に用いた。ヒトMEFLIN組換えタンパク質は哺乳類細胞発現系によって取得した。例えば、浮遊培養に馴化させたヒト胎児腎臓由来細胞株(Expi293F細胞、Thermo Fisher)にヒトMEFLIN遺伝子をリポフェクション法(lipofectamine2000, invitrogen)によって導入した。後の精製を可能にするためにMEFLIN遺伝子のカルボキシル末端部のGPIアンカー配列を除去し、ここにヒスチジンタグを導入した。遺伝子導入5~8日後の細胞培養液を回収し、フィルター(0.22マイクロメートル)処理後に、ヒスチジンタグ付きタンパク質を特異的に結合する担体(Ni Excel, GE Healthcare)を充填したカラムに供することによって培養液中のMEFLINタンパク質を結合させる。カラム洗浄後に、イミダゾール(500 mM)を含むバッファー液によって結合したMEFLINタンパク質を競合的に溶出させた。MEFLINタンパク質を含む溶出液から限外ろ過(リン酸緩衝液を用いた透析)によってイミダゾールを除去することによって最終的なヒトMEFLIN組換えタンパク質が得られた。精製されたヒトMEFLIN組換えタンパク質の純度は、タンパク質をSDS-PAGE電気泳動後Coomassie Brilliant Blue染色法に供することによって測定した。
WKYラット後肢の足底に組換えヒトMEFLINタンパク質(精製度95%)100μgを1回免疫した。2週間後に腸骨リンパ節に存在する形質細胞とミエローマ(SP2/0)細胞株を電気融合し、ハイブリドーマを作製した。培養液はHigh glucose DMEM(ナカライ)にHAT培地(MPB)とBM condimed H1(Roche)を添加し、ハイブリドーマのスクリーニングを行なった。
C57BL/6マウス尾根部筋肉に組換えヒトMEFLINタンパク質(精製度95%)50μgを1回免疫する以外は、ラット抗ヒトMEFLINモノクローナル抗体の作製と同様の方法でモノクローナル抗体を作製した。
組換えヒトMEFLINタンパク質を用いたELISA、hMEFLIN CHO細胞株を用いた免疫蛍光染色(IF)、hMEFLIN CHO細胞株を用いたフローサイトメトリー、hMeflin HEK293細胞株より作製した細胞溶解液を用いた免疫沈降、およびウェスタンブロッティングにより、各方法において抗原を認識するモノクローナル抗体を産生するハイブリドーマをスクリーニングした。
ヒト胎児腎(HEK293)細胞株にヒトMEFLINタンパク質またはその断片をコードするDNAが挿入されたpRP-CMVベクターをリポフェクション法により一過性に発現させ、細胞溶解液を作成した。断片は、図1または2に示される断片であり、C末端にG196タグおよびHisタグを有する。当該細胞溶解液を用いたSDS-PAGEを行い、泳動したタンパク質をPVDF膜へと転写した。当該PVDF膜を5%スキムミルク/PBSを用いて常温で1時間反応させた。1次抗体としては、各ハイブリドーマクローンの培養上清をPBSで50倍希釈したものを用いた。陽性対象においては、1次抗体として抗G196抗体を用いた。1次抗体存在下で当該PVDF膜とともに4℃で一晩反応させた。その後、PVDF膜を0.05%Tween/PBSで洗浄した。2次抗体としては、抗ラットまたは抗マウス抗体-セイヨウワサビペルオキシダーゼ(HRP)コンジュゲートを用いた。2次抗体は、PBSで1000倍希釈して、当該PVDF膜と常温で45分反応させた。0.05%Tween/PBSで洗浄後、抗体を標識するHRPをECL(GEヘルスケア)で発光させ、CCDイメージャー(Las4000、GEヘルスケア)で画像をコンピュータに取り込んだ。
F-12 Glutamax培養液(Gibco)に20mM HEPSと0.1%BSAを添加したアッセイ溶液中でhMeflin CHO細胞株又はmMeflin CHO細胞株を37℃、5%CO2で3時間培養した。各ハイブリドーマ上清をアッセイ溶液で10倍希釈し、上記CHO細胞株に添加し15分間培養した。PBS(4℃)で上記CHO細胞株を洗浄し、洗浄溶液(0.5M NaCl, 3%酢酸含有のMilliQ-H2O)で洗浄して、細胞膜上に結合している抗体を除去した。上記CHO細胞株を4%パラホルムアルデヒド(PFA)で10分反応させ固定し、次に0.1%TiritonX-100で5分反応させ、細胞膜透過化処理を行った。2次抗体としては抗ラットまたは抗マウス抗体-Alexa488コンジュゲートを用いた2次抗体をPBSで400倍希釈し、細胞膜透過化処理した細胞と常温で30分反応させた。PBSで洗浄後、DAPIと常温で5分反応させ、PBSで洗浄し、共焦点顕微鏡(LSM700、ZEISS)で画像を取得した。
肉腫患者またはがん腫患者から得られた病理組織標本から鏡検下で腫瘍成分が最も多い領域を選別し、MEFLIN(「ISLR」ともいう)の発現をIn situハイブリダイゼーション法(以下「ISH染色法」という)で調べた(WO2017/22472参照)。調べた組織の内、中倍率視野(20倍対物レンズ)で組織を観察し、肉腫組織に関しては腫瘍細胞の形態を有する細胞の20%以上がISLR陽性であるものをISLR陽性の腫瘍組織とし、それより少ないものを陰性の腫瘍組織としてグループ分けした。同様に、がん腫においてはがん間質に浸潤する線維芽細胞様形態を有する細胞の20%以上がISLR陽性であるものをISLR陽性の腫瘍組織とし、それより少ないものを陰性の腫瘍組織としてグループ分けした。細胞質の少なくとも一部においてシグナルが観察される場合には、ISLR陽性であるとした。
骨肉腫細胞株(HsOs1)、膵臓がん細胞株(BxPC-3)を皮下移植した担がんモデルマウスの腫瘍組織をサンプリングし、10%ホルマリンで固定後にパラフィンで包埋した。2μmの厚さに薄切しパラフィン包埋組織をスライドガラスに静置し、キシレンとエタノールにより脱パラフィンを行い、pH6.0の抗原賦活化液(Leica)により抗原の賦活化を行った。次に、ブロッキングを行った後に、抗MEFLINモノクローナル抗体と抗カテプシンK抗体(ab37259、abcam)を1次抗体として使用し、抗ラット抗体-Alexa488コンジュゲートと抗マウス抗体-Alexa594コンジュゲートを2次抗体として使用し(Invitrogen)、免疫蛍光二重染色法を行った。細胞内の核はDAPI染色を行った。MEFLIN(緑色)、カテプシンK(赤色)、核(青色)である。共焦点顕微鏡(LSM700、ZEISS)で画像を取得した。
がん腫患者から得られた病理組織標本から鏡検下で腫瘍成分が最も多い領域を選別し、MEFLIN(「ISLR」ともいう)の発現を免疫組織染色法(以下「IHC染色法」という)で調べた。調べた組織の内、中倍率視野(20倍対物レンズ)で組織を観察し、がん間質に浸潤する線維芽細胞様形態を有する細胞の20%以上がISLR陽性であるものをISLR陽性の腫瘍組織とし、それより少ないものを陰性の腫瘍組織としてグループ分けした。細胞質の少なくとも一部においてシグナルが観察される場合には、ISLR陽性であるとした。抗ラット又は抗マウス抗体-セイヨウワサビペルオキシダーゼ(HRP)コンジュゲートを2次抗体として使用した(ImmPRESS, VECTORS LABORATORIES)。
ウェスタンブロッティングによって各モノクローナル抗体と各ヒトMEFLINタンパク質断片との結合を確認した結果は図1に示される通りであった。図1に示されるように、クローン27-7から産生されるラットまたはマウス由来モノクローナル抗体(以下、単に「27-7抗体」という)は、ヒトMEFLINの1~343番目のアミノ酸領域には結合せず、1~399番目のアミノ酸領域に結合した。このことから、27-7抗体は、ヒトMEFLINの344~399番目のアミノ酸領域に結合することが示唆された。同様に、図1から、34-4抗体は232~343番目のアミノ酸領域に結合することが示唆され、;35-9抗体は1~146番目のアミノ酸領域することが示唆され;27-8抗体、41-10抗体、および46-3抗体はヒトMEFLINの344~399番目のアミノ酸領域に結合することが示唆された。また、図2に示されるように、3-2抗体、11-8抗体、12-2抗体、19-7抗体、および23-1抗体は、ヒトMEFLINの344~399番目のアミノ酸領域に結合することが示唆され;13-1抗体は147~231番目のアミノ酸領域に結合することが示唆され;16-5抗体、22-3抗体、および32-1抗体は1~146番目のアミノ酸領域に結合することが示唆された。
本実施例では、実施例1で作製した抗体と細胞傷害剤とのコンジュゲートを作製した。また、作製したコンジュゲートについて、担がんマウスモデルに投与して、抗腫瘍効果を確認した。
ハイブリドーマの高密度培養によって濃縮した抗体含有ハイブリドーマ上清からMono Spin(商標) L ProG(GL Sciense)を用いて抗体を精製した。1.0mM DTTによる還元処理を25℃で30分間行うことにより抗体のジスルフィド結合を切断し、露出したチオール基にVcMMAE(mc-vc-PAB-MMAE、MedChemExpress)またはmc-PEG12-vc-PABC-MMAEを常温で付加させ、限界ろ過によって精製した。VcMMAEは、CAS番号646502-53-6の下で登録される化合物である。VcMMAEは、モノメチルオーリスタチンE(MMAE)がp-アミノベンゾイルオキシカルボニル(PBAC)を介して、バリン-シトルリンリンカーと結合しており、カテプシンB等のプロテアーゼによってバリン-シトルリンが切断されるとMMAEが放出される仕組みとなっている。バリン-シトルリンリンカーは、マレイミドカプロイル基で修飾されており、マレイミド基を介して抗体のシステインが有するチオール基と反応することができる。mc-PEG12-vc-PABC-MMAEは、PEGが介在する以外は、VcMMAEと同様であり、MMAE放出のメカニズムもVcMMAEと同じである。薬物抗体比率(抗体1分子に対してコンジュゲートされる細胞傷害剤の個数、DAR)は、還元処理後の抗体のチオール基個数と、抗体にVcMMAEを付加させた後のチオール基個数の差を算出することによって決定した。具体的には、露出したチオール基に5,5’-ジチオビス(2-ニトロ安息香酸、同仁化学研究所)を反応させ、安定な5-メルカプト-2-ニトロ安息香酸を生成する。この生成したチオールの吸光度(λmax=412 nm、ε=1.55×104)からチオール基を定量する。
まず、ヒトMEFLIN発現HEK293細胞に対するADCの細胞傷害性を試験した。具体的には、hMeflin HEK293細胞株を1.0×104/ウェルでプレートに播種し、37℃、5%CO2で1日培養した。次に、各濃度のADCを添加し、2日培養し、MTT試験により細胞増殖能を評価した。細胞増殖能は、MMT試験においてサンプルを波長590nmにおける吸光度を吸光度計(POWERSCAN4、DS PHARMA BIOMEDICAL)で測定することにより評価した。また、hMeflin HEK293細胞株におけるMeflin発現量をFACS解析で評価した。以下、図中で単にクローン番号が記載されている場合には、モノクローナル抗体そのものが用いられたことを意味し、ADCの直前または直後にクローン番号が記載されている場合には、抗体が当該クローンであるADCが用いされたことを意味し、CTL ADCまたはアイソタイプコントロールIgG ADCは、抗体がアイソタイプコントロールIgG抗体であるADCが用いられたことを意味する。アイソタイプコントロール抗体としてラットIgG2a(BioLegend)を用いた。
上記と同様に、横紋筋肉腫(KYM-1)細胞株に対するADCの細胞傷害性を試験した。結果は図7に示されるとおりであった。図7に示されるように、横紋筋肉腫細胞株は、ヒトMEFLINタンパク質を強く発現していることがFACS解析から明らかであった。また、ADC27-7およびADC34-4が、KYM-1細胞株の増殖を有意に低下させた。
次に、担がんマウスモデルを用いてインビボでのADCによる細胞傷害性を試験した。KYM-1細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約100mm3前後の腫瘍体積を有するマウスに対してADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、KYM細胞株の移植によって形成された腫瘍のISH染色を行った。
さらに、内因性Meflin陽性骨肉腫(HsOs1)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約75mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、HsOs1細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト膵臓がん(BxPC-3)細胞株1.0×107個をメス ヌードマウス(BALB/cSlc nu/nu)の背部に皮下移植した(各群n=5)。約150mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、BxPC-3細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト肺がん(A549)細胞株1.0×107個をメス ヌードマウス(BALB/cSlc nu/nu)の背部に皮下移植した(各群n=5)。約100mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、A549細胞株の移植によって形成された腫瘍のISH染色を行った。
さらに、内因性Meflin陽性神経芽腫(NB-1)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約250mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、NB-1細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト大腸がん(DLD-1)細胞株1.0×107個をメス ヌードマウス(BALB/cSlc nu/nu)の背部に皮下移植した(各群n=5)。約200mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、DLD-1細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト胃がん(MKN45)細胞株1.0×107個をメス ヌードマウス(BALB/cSlc nu/nu)の背部に皮下移植した(各群n=5)。約200mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、MKN45細胞株の移植によって形成された腫瘍のISH染色を行った。
図15は、インターネット上に公開されたマウス膵臓に由来する細胞集団のシングルセルRNAシーケンスデータ(Tabula Muris)(非特許文献7参照)を解析した。縦軸、横軸にそれぞれ示すtSNE(t-distributed Stochastic Neighbor Embedding;t分布型確率的近傍埋め込み)は、多量のRNA発現量データから構成される多次元データを非線形変換により1次元まで圧縮して表した数値を表す。つまり、図15は、個々の細胞で発現する多量のRNA発現量データをtSNE1とtSNE2による2次元平面に展開することで、遺伝子発現プロファイルに基づいて細胞をクラスタリングすることができる。この手法では、例えば、丸印で示される個々の細胞がどのような細胞クラスターを形成しているのか理解することができる。これにより、当該クラスタリング手法では、特定因子が発現する細胞群と、他の特定因子が発現する細胞群が一致するかどうかを確認することが可能となり、これに加えて、当該細胞群がどの細胞種に対応するかを決定することも可能となる。図15は、マウス膵臓の膵星細胞(Pancreatic stellate cell)がMEFLINおよびカテプシンKの両者を発現することを示しており、マウス膵臓内において、MEFLIN陽性細胞集団とカテプシンK陽性細胞集団が一致することを示す(図中の矢印参照)。
図16は、インターネット上に公開されたマウス肺に由来する細胞集団のシングルセルRNAシーケンスデータ(Tabula Muris)(非特許文献7参照)を解析した。マウス肺の間質細胞(Stromal cell)にMEFLINとカテプシンKの両者が発現することを示しており、マウス肺において、MEFLIN陽性細胞集団とカテプシンK陽性細胞集団が一致することを示す(図中の矢印参照)。
(図中の矢印参照)。
図18は、骨肉腫細胞株(HsOs1)を皮下移植した担がんモデルマウスの腫瘍組織に対して抗MEFLINモノクローナル抗体と抗カテプシンK抗体を用いた免疫蛍光二重染色法を行った結果を示す。MEFLIN陽性細胞(緑色)の周りにカテプシンK(赤色)が分泌されていることを示す(矢印)。
図19は、膵臓がん細胞株(BxPC―3)を皮下移植した担がんモデルマウスの腫瘍組織に対して抗MEFLINモノクローナル抗体と抗カテプシンK抗体を用いた免疫蛍光二重染色法を行った結果を示す。MEFLIN陽性細胞(緑色)の周りにカテプシンK(赤色)が分泌されていることを示す(矢印)。
内因性Meflin陰性ヒト胆管がん(HuCCT1)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約250mm3前後の腫瘍体積を有するマウスに対して、ADCを尾静脈投与した(5mg/kg体重、4日間隔で計3回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、HuCCT1細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト膀胱がん(T24)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=4)。約300mm3前後の腫瘍体積を有するマウスに対して、ADCを尾静脈投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、T24細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト卵巣がん(OV-90)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約100mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、OV-90細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト食道がん(KYSE)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=5)。約200mm3前後の腫瘍体積を有するマウスに対して、ADCを腹腔内投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、KYSE細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陽性骨肉腫(HsOs1)細胞株1.0×107個をメスNOD SCIDマウスの背部に皮下移植した(各群n=4)。約250mm3前後の腫瘍体積を有するマウスに対して、ADCを尾静脈投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、HsOs1細胞株の移植によって形成された腫瘍のISH染色を行った。
内因性Meflin陰性ヒト乳がん(MCF-7)細胞株1.0×107個をメス ヌードマウス(BALB/cSlc nu/nu)の背部に皮下移植した(各群n=5)。約300mm3前後の腫瘍体積を有するマウスに対して、ADCを尾静脈投与した(5mg/kg体重、4日間隔で計5回投与)。腫瘍の長さ(L)及び幅(W)の測定は電動ノギスで測定し、体積はV=L×W2×0.52に従い計算した。また、MCF-7細胞株の移植によって形成された腫瘍のISH染色を行った。
対象サンプルには、ヒト膵臓がん手術検体の連続切片を用いた。ヒト膵臓がん手術検体の連続切片に対して抗MEFLIN抗体を用いた免疫組織染色を実施し、ヒト組織に対して抗MEFLIN抗体を用いることでMEFLINの発現を確認した。また、同連続切片に対してIn situハイブリダイゼーションによるヒトMEFLINのmRNAの発現を確認した。
Claims (18)
- MEFLINに結合する抗体と細胞傷害剤との抗体-薬物コンジュゲート(ADC)を含む、がんを処置することに用いるための医薬組成物。
- 前記抗体は、インターナライズ活性を有する、請求項1に記載の医薬組成物。
- ADCが、抗体と薬物とがリンカーを介して連結されたADCであり、リンカーが、細胞内で開裂される切断部位を有する、請求項1または2に記載の医薬組成物。
- MEFLINに結合する抗体であって、以下からなる群から選択される抗体:
(1A)配列番号1に記載のアミノ酸配列を有する重鎖CDR1、配列番号2に記載のアミノ酸配列を有する重鎖CDR2、および配列番号3に記載のアミノ酸配列を有する重鎖CDR3を含む重鎖可変領域と、配列番号4に記載のアミノ酸配列を有する軽鎖CDR1、配列番号5に記載のアミノ酸配列を有する軽鎖CDR2、および配列番号6に記載のアミノ酸配列を有する軽鎖CDR3を含む軽鎖可変領域とを有する抗体;
(1B)配列番号7に記載のアミノ酸配列を有する重鎖可変領域と、配列番号8に記載のアミノ酸配列を有する軽鎖可変領域とを有する抗体;
(1C)上記(1B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(1D)上記(1B)の抗体とMEFLINタンパク質上で重なるエピトープに結合する抗体;
(2A)配列番号9に記載の重鎖CDR1と、配列番号10に記載の重鎖CDR2と、配列番号11に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号12に記載の軽鎖CDR1と、配列番号13に記載の軽鎖CDR2と、配列番号14に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(2B)配列番号15に記載の重鎖可変領域と、配列番号16に記載の軽鎖可変領域を有する抗体;
(2C)上記(2B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(2D)上記(2B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(3A)配列番号17に記載の重鎖CDR1と、配列番号18に記載の重鎖CDR2と、配列番号19に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号20に記載の軽鎖CDR1と、配列番号21に記載の軽鎖CDR2と、配列番号22に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(3B)配列番号23に記載の重鎖可変領域と、配列番号24に記載の軽鎖可変領域を有する抗体;
(3C)上記(3B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(3D)上記(3B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(4A)配列番号25に記載の重鎖CDR1と、配列番号26に記載の重鎖CDR2と、配列番号27に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号28に記載の軽鎖CDR1と、配列番号29に記載の軽鎖CDR2と、配列番号30に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(4B)配列番号31に記載の重鎖可変領域と、配列番号32に記載の軽鎖可変領域を有
する抗体;
(4C)上記(4B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(4D)上記(4B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(5A)配列番号33に記載の重鎖CDR1と、配列番号34に記載の重鎖CDR2と、配列番号35に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号36に記載の軽鎖CDR1と、配列番号37に記載の軽鎖CDR2と、配列番号38に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(5B)配列番号39に記載の重鎖可変領域と、配列番号40に記載の軽鎖可変領域を有する抗体;
(5C)上記(5B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(5D)上記(5B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(6A)配列番号41に記載の重鎖CDR1と、配列番号42に記載の重鎖CDR2と、配列番号43に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号44に記載の軽鎖CDR1と、配列番号45に記載の軽鎖CDR2と、配列番号46に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(6B)配列番号47に記載の重鎖可変領域と、配列番号48に記載の軽鎖可変領域を有する抗体;
(6C)上記(6B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(6D)上記(6B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(7A)配列番号49に記載の重鎖CDR1と、配列番号50に記載の重鎖CDR2と、配列番号51に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号52に記載の軽鎖CDR1と、配列番号53に記載の軽鎖CDR2と、配列番号54に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(7B)配列番号55に記載の重鎖可変領域と、配列番号56に記載の軽鎖可変領域を有する抗体;
(7C)上記(7B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(7D)上記(7B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(8A)配列番号57に記載の重鎖CDR1と、配列番号58に記載の重鎖CDR2と、配列番号59に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号60に記載の軽鎖CDR1と、配列番号61に記載の軽鎖CDR2と、配列番号62に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(8B)配列番号63に記載の重鎖可変領域と、配列番号64に記載の軽鎖可変領域を有する抗体;
(8C)上記(8B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(8D)上記(8B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(9A)配列番号65に記載の重鎖CDR1と、配列番号66に記載の重鎖CDR2と、
配列番号67に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号68に記載の軽鎖CDR1と、配列番号69に記載の軽鎖CDR2と、配列番号70に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(9B)配列番号71に記載の重鎖可変領域と、配列番号72に記載の軽鎖可変領域を有する抗体;
(9C)上記(9B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(9D)上記(9B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(10A)配列番号73に記載の重鎖CDR1と、配列番号74に記載の重鎖CDR2と、配列番号75に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号76に記載の軽鎖CDR1と、配列番号77に記載の軽鎖CDR2と、配列番号78に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(10B)配列番号79に記載の重鎖可変領域と、配列番号80に記載の軽鎖可変領域を有する抗体;
(10C)上記(10B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(10D)上記(10B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(11A)配列番号81に記載の重鎖CDR1と、配列番号82に記載の重鎖CDR2と、配列番号83に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号84に記載の軽鎖CDR1と、配列番号85に記載の軽鎖CDR2と、配列番号86に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(11B)配列番号87に記載の重鎖可変領域と、配列番号88に記載の軽鎖可変領域を有する抗体;
(11C)上記(11B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(11D)上記(11B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(12A)配列番号89に記載の重鎖CDR1と、配列番号90に記載の重鎖CDR2と、配列番号91に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号92に記載の軽鎖CDR1と、配列番号93に記載の軽鎖CDR2と、配列番号94に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(12B)配列番号95に記載の重鎖可変領域と、配列番号96に記載の軽鎖可変領域を有する抗体;
(12C)上記(12B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(12D)上記(12B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(13A)配列番号97に記載の重鎖CDR1と、配列番号98に記載の重鎖CDR2と、配列番号99に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号100に記載の軽鎖CDR1と、配列番号101に記載の軽鎖CDR2と、配列番号102に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(13B)配列番号103に記載の重鎖可変領域と、配列番号104に記載の軽鎖可変領域を有する抗体;
(13C)上記(13B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(13D)上記(13B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(14A)配列番号105に記載の重鎖CDR1と、配列番号106に記載の重鎖CDR2と、配列番号107に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号108に記載の軽鎖CDR1と、配列番号109に記載の軽鎖CDR2と、配列番号110に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(14B)配列番号111に記載の重鎖可変領域と、配列番号112に記載の軽鎖可変領域を有する抗体;
(14C)上記(14B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および、
(14D)上記(14B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(15A)配列番号113に記載の重鎖CDR1と、配列番号114に記載の重鎖CDR2と、配列番号115に記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号116に記載の軽鎖CDR1と、配列番号117に記載の軽鎖CDR2と、配列番号118に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(15B)配列番号119に記載の重鎖可変領域と、配列番号120に記載の軽鎖可変領域を有する抗体;
(15C)上記(15B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(15D)上記(15B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;
(16A)配列番号121に記載の重鎖CDR1と、配列番号122に記載の重鎖CDR2と、配列番号123記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号124に記載の軽鎖CDR1と、配列番号125に記載の軽鎖CDR2と、配列番号126に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(16B)配列番号127に記載の重鎖可変領域と、配列番号128に記載の軽鎖可変領域を有する抗体;
(16C)上記(16B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(16D)上記(16B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体;並びに
(17A)配列番号129に記載の重鎖CDR1と、配列番号130に記載の重鎖CDR2と、配列番号131記載の重鎖CDR3を含む、重鎖可変領域と、
配列番号132に記載の軽鎖CDR1と、配列番号133に記載の軽鎖CDR2と、配列番号134に記載の軽鎖CDR3を含む、軽鎖可変領域と
を有する抗体;
(17B)配列番号135に記載の重鎖可変領域と、配列番号136に記載の軽鎖可変領域を有する抗体;
(17C)上記(17B)の抗体とMEFLINタンパク質との結合に関して競合する抗体;および
(17D)上記(17B)の抗体とMEFLINタンパク質上の重なるエピトープに結合する抗体。 - 請求項4に記載の抗体と、細胞傷害剤との抗体-薬物コンジュゲート(ADC)を含む、医薬組成物。
- がんを処置することに用いるための、請求項5に記載の医薬組成物。
- がんが、肉腫である、請求項1~3および6のいずれか一項に記載の医薬組成物。
- がんが、MEFLIN陽性である肉腫である、請求項7に記載の医薬組成物。
- 肉腫が、粘膜線維肉腫、悪性線維性組織球腫、脂肪肉腫、平滑筋肉腫、横紋筋肉腫、神経芽腫、悪性末梢神経鞘腫瘍、ユーイング肉腫、類上皮肉腫、明細胞肉腫、滑膜肉腫、および骨肉腫からなる群から選択される肉腫である、請求項7または8に記載の医薬組成物。
- がんが、癌腫である、請求項1~3および6のいずれか一項に記載の医薬組成物。
- がんが、乳がん、膵臓がん、肺がん、大腸がん、胃がん、胆管がん、卵巣がん、膀胱がん、および食道がんからなる群から選択されるがんである、請求項1~3および6のいずれか一項に記載の医薬組成物。
- がんが、MEFLIN陰性であり、当該がん周囲の間質がMEFLIN陽性の細胞を含む、請求項1~3、6、10、および11のいずれか一項に記載の医薬組成物。
- 抗体が、インターナライズ活性を有しない、請求項10~12のいずれか一項に記載の医薬組成物。
- 抗体が、インターナライズ活性を有する、請求項7~9のいずれか一項に記載の医薬組成物。
- ADCが、抗体と薬物とがリンカーを介して連結されたADCであり、リンカーが、開裂性リンカーである、請求項13に記載の医薬組成物。
- リンカーが、カテプシンKにより開裂する開裂性リンカーである、請求項15に記載の医薬組成物。
- リンカーが、バリン-シトルリンのジペプヂドを含み、カテプシンK存在下で切断される、請求項16に記載の医薬組成物。
- リンカーが、非開裂性リンカーである、上記のいずれかに記載の医薬組成物。
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP21750161.8A EP4101472A1 (en) | 2020-02-03 | 2021-02-03 | Anti-meflin antibody for use in treatment cancer in subject having cancer, and pharmaceutical composition including said antibody |
CN202180012327.9A CN115297890A (zh) | 2020-02-03 | 2021-02-03 | 用于对患癌的对象处置癌的抗meflin抗体和含有该抗体的药物组合物 |
AU2021217624A AU2021217624A1 (en) | 2020-02-03 | 2021-02-03 | Anti-MEFLIN antibody for use in treatment of cancer in subject having cancer, and pharmaceutical composition comprising the antibody |
US17/759,952 US20230084099A1 (en) | 2020-02-03 | 2021-02-03 | Anti-meflin antibody for use in treatment of cancer in subject having cancer, and pharmaceutical composition comprising the antibody |
JP2021575825A JPWO2021157601A1 (ja) | 2020-02-03 | 2021-02-03 | |
CA3168866A CA3168866A1 (en) | 2020-02-03 | 2021-02-03 | Anti-meflin antibody for use in treatment of cancer in subject having cancer, and pharmaceutical composition comprising the antibody |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2020016535 | 2020-02-03 | ||
JP2020-016535 | 2020-02-03 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2021157601A1 true WO2021157601A1 (ja) | 2021-08-12 |
Family
ID=77200605
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2021/003851 WO2021157601A1 (ja) | 2020-02-03 | 2021-02-03 | がんを有する対象においてがんを処置することに用いるための抗meflin抗体、および当該抗体を含む医薬組成物 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20230084099A1 (ja) |
EP (1) | EP4101472A1 (ja) |
JP (1) | JPWO2021157601A1 (ja) |
CN (1) | CN115297890A (ja) |
AU (1) | AU2021217624A1 (ja) |
CA (1) | CA3168866A1 (ja) |
WO (1) | WO2021157601A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023172879A3 (en) * | 2022-03-07 | 2023-11-16 | Nkarta, Inc. | Multiplex gene edited cells for cd70-directed cancer immunotherapy |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116693682B (zh) * | 2023-06-02 | 2024-04-19 | 涌源合生科技(深圳)有限公司 | 抗Tau蛋白抗体及其制备方法和用途 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017022472A1 (ja) | 2015-08-03 | 2017-02-09 | 国立大学法人名古屋大学 | 未分化間葉系幹細胞マーカー及びその用途 |
JP2019500327A (ja) * | 2015-11-30 | 2019-01-10 | アッヴィ・インコーポレイテッド | 抗huLRRC15抗体薬物コンジュゲート及びその使用方法 |
JP2019501124A (ja) * | 2015-11-30 | 2019-01-17 | アッヴィ・インコーポレイテッド | 抗huLRRC15抗体薬物コンジュゲート及びその使用方法 |
WO2019159825A1 (ja) | 2018-02-14 | 2019-08-22 | 国立大学法人名古屋大学 | 抗pd-1抗体/抗pd-l1抗体療法の効果を予測するバイオマーカー |
-
2021
- 2021-02-03 WO PCT/JP2021/003851 patent/WO2021157601A1/ja unknown
- 2021-02-03 CA CA3168866A patent/CA3168866A1/en active Pending
- 2021-02-03 CN CN202180012327.9A patent/CN115297890A/zh active Pending
- 2021-02-03 JP JP2021575825A patent/JPWO2021157601A1/ja active Pending
- 2021-02-03 US US17/759,952 patent/US20230084099A1/en active Pending
- 2021-02-03 EP EP21750161.8A patent/EP4101472A1/en active Pending
- 2021-02-03 AU AU2021217624A patent/AU2021217624A1/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017022472A1 (ja) | 2015-08-03 | 2017-02-09 | 国立大学法人名古屋大学 | 未分化間葉系幹細胞マーカー及びその用途 |
JP2019500327A (ja) * | 2015-11-30 | 2019-01-10 | アッヴィ・インコーポレイテッド | 抗huLRRC15抗体薬物コンジュゲート及びその使用方法 |
JP2019501124A (ja) * | 2015-11-30 | 2019-01-17 | アッヴィ・インコーポレイテッド | 抗huLRRC15抗体薬物コンジュゲート及びその使用方法 |
WO2019159825A1 (ja) | 2018-02-14 | 2019-08-22 | 国立大学法人名古屋大学 | 抗pd-1抗体/抗pd-l1抗体療法の効果を予測するバイオマーカー |
Non-Patent Citations (13)
Title |
---|
"GenBank", Database accession no. BAA85970.1 |
CAS , no. 646502-53-6 |
CRISAN M. ET AL., CELL STEM CELL., vol. 3, 2008, pages 301 - 313 |
ENOMOTO, ATSUSHI: "WS-1-01-4 Identification and nature of "tumor-suppressive fibroblasts" in cancer stroma", PROGRAMS AND ABSTRACTS OF THE 27TH ANNUAL MEETING OF THE JAPANESE BREAST CANCER SOCIETY; JULY 11-13, 2019, 30 November 2018 (2018-11-30) - 13 July 2019 (2019-07-13), JP, pages 274, XP009538912 * |
HARA A. ET AL., CIRC. RES., vol. 125, 2019, pages 414 - 430 |
KOBAYASHI H. ET AL., NAT REV GASTROENTEROL HEPATOL., vol. 16, 2019, pages 282 - 295 |
MAEDA K. ET AL., SCI. REP., vol. 6, 2016, pages 22288 |
MIZUTANI Y. ET AL., CANCER RES., vol. 79, 2019, pages 5367 - 5381 |
MIZUTANI YASUYUKI, KOBAYASHI HIROKI, IIDA TADASHI, ASAI NAOYA, MASAMUNE ATSUSHI, HARA AKITOSHI, ESAKI NOBUTOSHI, USHIDA KAORI, MII: "Meflin-Positive Cancer-Associated Fibroblasts Inhibit Pancreatic Carcinogenesis", CANCER RESEARCH, vol. 79, no. 20, 2019, pages 5367 - 5381, XP055845782, ISSN: 0008-5472 * |
NAGASAWA, A. ET AL., GENOMICS, vol. 44, 1977, pages 273 - 279 |
PURCELL, J. W. ET AL.: "LRRC15 Is a Novel Mesenchymal Protein and Stromal Target for Antibody-Drug Conjugates", CANCER RES, vol. 78, no. 14, 2018, pages 4059 - 4072, XP055745280, ISSN: 0008-5472, DOI: 10.1158/0008-5472.CAN-18-0327 * |
SCHAUM, N. ET AL., NATURE, vol. 562, no. 7727, 2018, pages 367 - 372 |
SZOT, C. ET AL.: "Tumor stroma-targeted antibody- drug conjugate triggers localized anticancer drug release", J CLIN INVEST, vol. 128, no. 7, 2018, pages 2927 - 2943, XP055724470, ISSN: 0021-9738, DOI: 10.1172/JCI120481 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023172879A3 (en) * | 2022-03-07 | 2023-11-16 | Nkarta, Inc. | Multiplex gene edited cells for cd70-directed cancer immunotherapy |
Also Published As
Publication number | Publication date |
---|---|
JPWO2021157601A1 (ja) | 2021-08-12 |
US20230084099A1 (en) | 2023-03-16 |
CA3168866A1 (en) | 2021-08-12 |
AU2021217624A1 (en) | 2022-09-01 |
CN115297890A (zh) | 2022-11-04 |
EP4101472A1 (en) | 2022-12-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10640571B2 (en) | Human monoclonal antibodies specific for glypican-3 and use thereof | |
US9409994B2 (en) | High-affinity monoclonal antibodies to glypican-3 and use thereof | |
WO2017196847A1 (en) | Variable new antigen receptor (vnar) antibodies and antibody conjugates targeting tumor and viral antigens | |
JP7032425B2 (ja) | Muc1に特異的に結合する抗体及びその用途 | |
CN109988240B (zh) | 抗gpc-3抗体及其用途 | |
WO2021157601A1 (ja) | がんを有する対象においてがんを処置することに用いるための抗meflin抗体、および当該抗体を含む医薬組成物 | |
JP2021535209A (ja) | ソルチリン受容体を標的化し血管擬態を阻害するための方法及び化合物 | |
US20180002439A1 (en) | Anti-mesothelin antibodies and uses thereof | |
US9150655B2 (en) | Anti-C-met antibody and methods of use thereof | |
WO2018203517A1 (ja) | プラスミンにより切断可能な抗不溶性フィブリン抗体と薬物とのコンジュゲート | |
CN111201244A (zh) | 特异性结合到水通道蛋白3(aqp3)的细胞外域的抗aqp3单克隆抗体及其用途 | |
JP2019512548A (ja) | ポドカリキシン及びtra関連抗体、調製方法、並びに抗癌治療剤としての使用 | |
WO2021251460A1 (ja) | Cafを有するがんを処置するための組成物 | |
WO2021251459A1 (ja) | ヒト化抗gpc-1抗体 | |
US20230365676A1 (en) | Cd33 antibodies | |
JP5939855B2 (ja) | トランスフェリン受容体抗体 | |
KR102090444B1 (ko) | 항-icam4 항체 및 icam4 발현 세포와 관련된 질병의 진단 및 치료용 조성물 | |
WO2020158959A1 (ja) | 抗mc16抗体 | |
JP2020520230A (ja) | Paufタンパク質に特異的に結合する抗体及びその用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21750161 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2021575825 Country of ref document: JP Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: 3168866 Country of ref document: CA |
|
ENP | Entry into the national phase |
Ref document number: 2021217624 Country of ref document: AU Date of ref document: 20210203 Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2021750161 Country of ref document: EP Effective date: 20220905 |