JP5899310B2 - c−KIT抗体およびその使用 - Google Patents
c−KIT抗体およびその使用 Download PDFInfo
- Publication number
- JP5899310B2 JP5899310B2 JP2014510362A JP2014510362A JP5899310B2 JP 5899310 B2 JP5899310 B2 JP 5899310B2 JP 2014510362 A JP2014510362 A JP 2014510362A JP 2014510362 A JP2014510362 A JP 2014510362A JP 5899310 B2 JP5899310 B2 JP 5899310B2
- Authority
- JP
- Japan
- Prior art keywords
- kit
- seq
- antibody
- amino acid
- human
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000282414 Homo sapiens Species 0.000 claims description 78
- 230000027455 binding Effects 0.000 claims description 37
- 238000000034 method Methods 0.000 claims description 26
- 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 15
- 239000012634 fragment Substances 0.000 claims description 10
- 239000000427 antigen Substances 0.000 claims description 7
- 102000036639 antigens Human genes 0.000 claims description 7
- 108091007433 antigens Proteins 0.000 claims description 7
- 102000016971 Proto-Oncogene Proteins c-kit Human genes 0.000 claims description 4
- 108010014608 Proto-Oncogene Proteins c-kit Proteins 0.000 claims description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims 9
- 210000004027 cell Anatomy 0.000 description 88
- 150000001413 amino acids Chemical group 0.000 description 63
- 206010028980 Neoplasm Diseases 0.000 description 40
- 238000011282 treatment Methods 0.000 description 37
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 30
- 210000004881 tumor cell Anatomy 0.000 description 28
- 201000011510 cancer Diseases 0.000 description 21
- 230000015556 catabolic process Effects 0.000 description 21
- 238000006731 degradation reaction Methods 0.000 description 21
- 238000000338 in vitro Methods 0.000 description 19
- 208000035475 disorder Diseases 0.000 description 17
- 230000001419 dependent effect Effects 0.000 description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 201000010099 disease Diseases 0.000 description 13
- 230000004565 tumor cell growth Effects 0.000 description 13
- 239000002299 complementary DNA Substances 0.000 description 12
- 230000026731 phosphorylation Effects 0.000 description 12
- 238000006366 phosphorylation reaction Methods 0.000 description 11
- 102000043136 MAP kinase family Human genes 0.000 description 10
- 108091054455 MAP kinase family Proteins 0.000 description 10
- 239000002246 antineoplastic agent Substances 0.000 description 10
- 238000003556 assay Methods 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 238000001262 western blot Methods 0.000 description 10
- 210000000170 cell membrane Anatomy 0.000 description 9
- 239000003814 drug Substances 0.000 description 9
- 238000011534 incubation Methods 0.000 description 9
- 208000032839 leukemia Diseases 0.000 description 9
- 239000011780 sodium chloride Substances 0.000 description 9
- 230000001225 therapeutic effect Effects 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 230000000903 blocking effect Effects 0.000 description 8
- 239000000872 buffer Substances 0.000 description 8
- 239000013604 expression vector Substances 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 239000003446 ligand Substances 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 7
- 238000010790 dilution Methods 0.000 description 7
- 239000012895 dilution Substances 0.000 description 7
- 201000011243 gastrointestinal stromal tumor Diseases 0.000 description 7
- 108020004707 nucleic acids Proteins 0.000 description 7
- 102000039446 nucleic acids Human genes 0.000 description 7
- 150000007523 nucleic acids Chemical class 0.000 description 7
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 7
- 208000006168 Ewing Sarcoma Diseases 0.000 description 6
- 206010041067 Small cell lung cancer Diseases 0.000 description 6
- 238000000423 cell based assay Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 201000001441 melanoma Diseases 0.000 description 6
- 239000008194 pharmaceutical composition Substances 0.000 description 6
- 230000036962 time dependent Effects 0.000 description 6
- 208000008839 Kidney Neoplasms Diseases 0.000 description 5
- 206010029260 Neuroblastoma Diseases 0.000 description 5
- 206010038389 Renal cancer Diseases 0.000 description 5
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 5
- 229960004316 cisplatin Drugs 0.000 description 5
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 5
- 229960005420 etoposide Drugs 0.000 description 5
- 201000010982 kidney cancer Diseases 0.000 description 5
- 239000012139 lysis buffer Substances 0.000 description 5
- 208000000587 small cell lung carcinoma Diseases 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 4
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- 108060003951 Immunoglobulin Proteins 0.000 description 4
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 4
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 4
- 206010035226 Plasma cell myeloma Diseases 0.000 description 4
- 108010076504 Protein Sorting Signals Proteins 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 239000000306 component Substances 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 4
- 102000018358 immunoglobulin Human genes 0.000 description 4
- 238000000099 in vitro assay Methods 0.000 description 4
- 238000005462 in vivo assay Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 201000000050 myeloid neoplasm Diseases 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 229940124597 therapeutic agent Drugs 0.000 description 4
- 239000012130 whole-cell lysate Substances 0.000 description 4
- 101100002068 Bacillus subtilis (strain 168) araR gene Proteins 0.000 description 3
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 3
- 101710177504 Kit ligand Proteins 0.000 description 3
- 102100020880 Kit ligand Human genes 0.000 description 3
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 238000001042 affinity chromatography Methods 0.000 description 3
- 101150044616 araC gene Proteins 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 210000002798 bone marrow cell Anatomy 0.000 description 3
- 208000035269 cancer or benign tumor Diseases 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 229960000684 cytarabine Drugs 0.000 description 3
- 229940127089 cytotoxic agent Drugs 0.000 description 3
- 229960003901 dacarbazine Drugs 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000010494 dissociation reaction Methods 0.000 description 3
- 230000005593 dissociations Effects 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 210000004962 mammalian cell Anatomy 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 3
- 102000027426 receptor tyrosine kinases Human genes 0.000 description 3
- 108091008598 receptor tyrosine kinases Proteins 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 239000012679 serum free medium Substances 0.000 description 3
- 239000012089 stop solution Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 201000009030 Carcinoma Diseases 0.000 description 2
- 238000003734 CellTiter-Glo Luminescent Cell Viability Assay Methods 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 238000012413 Fluorescence activated cell sorting analysis Methods 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101001008874 Homo sapiens Mast/stem cell growth factor receptor Kit Proteins 0.000 description 2
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 2
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 2
- 239000005517 L01XE01 - Imatinib Substances 0.000 description 2
- 239000002147 L01XE04 - Sunitinib Substances 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 241000282567 Macaca fascicularis Species 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- 108010004729 Phycoerythrin Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 241000508269 Psidium Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 206010039491 Sarcoma Diseases 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000000556 agonist Substances 0.000 description 2
- 239000004037 angiogenesis inhibitor Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229960000074 biopharmaceutical Drugs 0.000 description 2
- 201000000053 blastoma Diseases 0.000 description 2
- 239000007975 buffered saline Substances 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 238000000006 cell growth inhibition assay Methods 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 230000002759 chromosomal effect Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 201000008184 embryoma Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 238000009650 gentamicin protection assay Methods 0.000 description 2
- 230000011132 hemopoiesis Effects 0.000 description 2
- 210000003630 histaminocyte Anatomy 0.000 description 2
- 102000055152 human KIT Human genes 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- KTUFNOKKBVMGRW-UHFFFAOYSA-N imatinib Chemical compound C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 KTUFNOKKBVMGRW-UHFFFAOYSA-N 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000003061 melanogenesis Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- LBWFXVZLPYTWQI-IPOVEDGCSA-N n-[2-(diethylamino)ethyl]-5-[(z)-(5-fluoro-2-oxo-1h-indol-3-ylidene)methyl]-2,4-dimethyl-1h-pyrrole-3-carboxamide;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C LBWFXVZLPYTWQI-IPOVEDGCSA-N 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000001613 neoplastic effect Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 238000003566 phosphorylation assay Methods 0.000 description 2
- 229940012957 plasmin Drugs 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000000069 prophylactic effect Effects 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 230000021595 spermatogenesis Effects 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- 108010032595 Antibody Binding Sites Proteins 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010051066 Gastrointestinal stromal tumour Diseases 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 101000716729 Homo sapiens Kit ligand Proteins 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 230000004163 JAK-STAT signaling pathway Effects 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102100027754 Mast/stem cell growth factor receptor Kit Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102000016979 Other receptors Human genes 0.000 description 1
- 102000038030 PI3Ks Human genes 0.000 description 1
- 108091007960 PI3Ks Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 230000010799 Receptor Interactions Effects 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 206010039897 Sedation Diseases 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000009833 antibody interaction Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 230000035578 autophosphorylation Effects 0.000 description 1
- 238000013357 binding ELISA Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000012200 cell viability kit Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 239000002895 emetic Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000010437 erythropoiesis Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 229940080856 gleevec Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 102000005396 glutamine synthetase Human genes 0.000 description 1
- 108020002326 glutamine synthetase Proteins 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 102000055151 human KITLG Human genes 0.000 description 1
- 229960002411 imatinib Drugs 0.000 description 1
- 229960003685 imatinib mesylate Drugs 0.000 description 1
- YLMAHDNUQAMNNX-UHFFFAOYSA-N imatinib methanesulfonate Chemical compound CS(O)(=O)=O.C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 YLMAHDNUQAMNNX-UHFFFAOYSA-N 0.000 description 1
- 230000037189 immune system physiology Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 229940125721 immunosuppressive agent Drugs 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940127084 other anti-cancer agent Drugs 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 239000002464 receptor antagonist Substances 0.000 description 1
- 229940044551 receptor antagonist Drugs 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 230000036280 sedation Effects 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 238000009097 single-agent therapy Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- WINHZLLDWRZWRT-ATVHPVEESA-N sunitinib Chemical compound CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C WINHZLLDWRZWRT-ATVHPVEESA-N 0.000 description 1
- 229960001796 sunitinib Drugs 0.000 description 1
- 229960002812 sunitinib malate Drugs 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229940034785 sutent Drugs 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
- 238000007693 zone electrophoresis Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
- A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Oncology (AREA)
- General Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Hematology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
抗ヒトc−Kit mAb、CK6のCDRのアミノ酸配列を表1に示す。
抗c−Kit mAbの重鎖可変領域および軽鎖可変領域をコードするcDNA配列は当該技術分野において公知の手順に従って発現ベクター中でクローニングするためにPCRにより増幅され得る。例えば、重鎖可変領域(例えば配列番号8)をコードするcDNAは、フレームにおいて、ベクターpEE6.1(Lonza Biologics plc,Slough,Berkshire,UK)中のヒト免疫グロブリン重鎖ガンマ定常領域をコードするcDNA配列に融合され得る。全ヒト軽鎖(例えば配列番号16)をコードするcDNAはベクターPEE12.1(Lonza Biologics PLC,Slough,Berkshire,UK)中で直接クローニングされ得る。操作された免疫グロブリン発現ベクターは、エレクトロポレーションおよびグルタミン合成酵素選択培地中で培養された形質転換体によりNS0骨髄腫細胞中で安定にトランスフェクトされ得る。安定なクローンが抗ヒトc−Kit特異的結合ELISAにより抗体発現のためにスクリーニングされ得る。陽性クローンは、スピナーフラスコまたはバイオリアクタ中で抗体産生のために無血清培地培養物中で培養され得る。完全長IgG1抗体は、タンパク質親和性クロマトグラフィー(Poros A,PerSeptive Biosystems Inc.,Foster City,CA)により精製され得、中性緩衝生理食塩水中で溶出され得る。
精製した抗c−Kit mAbのインビトロでのc−Kit結合および遮断活性は、例えば酵素結合免疫吸着測定法(ELISA)などの当該技術分野において公知の免疫学的、定量的技術を使用して決定できる。簡潔に述べると、96ウェルマイクロタイタープレートを、室温(20〜25℃)にて1時間、100μl/ウェルの1μg/ml組換えヒトc−Kit(rh−c−Kit)タンパク質(R&D Systems,Minneapolis,MN)でコーティングする。洗浄後、ウェルを3%PBS/ミルクで遮断する。典型的に1.5μg/mlまたは100nMで開始する、精製した抗c−Kit mAbの異なる希釈物を次いでrh−c−Kitでコーティングしたウェルに加え、室温にて2時間インキュベートさせる。数回洗浄した(3〜5回)後、抗ヒトIgG−HRPがコンジュゲートした抗体(Jackson ImmunoResearch,West Grove,PA)の1:1000希釈物を室温にて1時間、プレートに加える。プレートを洗浄し、HRP基質を各ウェルに加え、青色が発生するまでインキュベートする。停止溶液を加え、吸光度を450nmにて測定する。上記のようにELISAにおいてrh−c−Kitタンパク質への結合についてモノクローナル抗体CK6を試験した。結果は、約3×10−11MのEC50値でCK6がrh−c−Kitに結合することを示す。
rh−c−kitに対する抗c−Kitモノクローナル抗体の結合キネティクスは、当該技術分野において公知の方法に従ってBIAcore(登録商標)3000またはBIAcore(登録商標)T100(GE HealthCare,Piscataway,NJ)などのSPRバイオセンサにより決定できる。本質的に、抗cKitモノクローナル抗体は、BIAcore(登録商標)T100および結合親和性測定の間のランニング緩衝液としてHBS−EP(0.01M HEPES−pH7.4、0.15mM NaCl、3mM EDTAおよび0.005%(v/v)界面活性剤P20)を使用してSPRで評価できる。測定は25℃で実施する。rh−c−kit(pH5.0)の約120反応単位(RU)は標準的なアミンカップリング手順を使用してCM5チップ上で固定化できる。mAbは、約30μL/分の流速にて、固定化したrh−c−kitに対して180秒間注入でき、続いてHBS−EPを使用して900秒解離する。抗c−Kitモノクローナル抗体の濃度は、2倍希釈の勾配であってもよく、1回の勾配当たり少なくとも3つの濃度であり、1回の実験当たり3回の勾配である。解離後、rh−c−kit表面の再生は、30μ/分にて50mMのHClの2回の30秒の注入により達成され得る。BIAcore(登録商標)T100評価ソフトウェアは速度定数を決定するために使用できる。親和定数、KDは速度定数Koff/Konの比から算出できる。相互作用の「Kon、M−1s−1」および「Koff、s−1」速度は抗体/受容体相互作用の親和性(KD、M)を決定するために使用する。
抗ヒトc−Kit mAbの種結合特異性は、ELISAにおいてヒト、カニクイザルおよびマウスc−Kitタンパク質に対する抗体の反応性を測定することにより決定できる。簡潔に述べると、96ウェルマイクロタイタープレートを、100μl/ウェルの1μg/ml組換えヒトc−Kitタンパク質(rh−c−Kit)または市販(R&D Systems)で得られる組換えマウスc−Kit(rm−c−Kit;Gln26−Thr519)またはカニクイザルc−Kit(組換えて調製したcyno−c−Kit;Met1−His549)で室温にて1時間、コーティングする。洗浄後、ウェルを3%PBS/ミルクで遮断する。ヒト抗ヒト−c−Kit mAbの連続希釈物を次いでc−Kitでコーティングしたウェルに加え、室温にて2時間インキュベートする。数回洗浄した後、結合したヒト抗体を検出するために、抗ヒトIgG−HRPがコンジュゲートした抗体の1:1000希釈物を室温にて1時間プレートに加える。プレートを洗浄し、HRP基質を各ウェルに加え、青色が発生するまでインキュベートする。停止溶液を加え、吸光度を450nmにて測定する。その結果は、CK6が(5.7×10−11MのEC50で)rh−c−Kit、カニクイザルc−Kitに結合するが、rm−c−Kitに有意に結合しないことを示す(表4)。
ヒトc−Kit発現細胞に対して向けられるc−Kit mAbの中和活性を測定するためにインビトロの細胞ベースアッセイを使用できる。c−Kit mAbにより誘導される内在化に基づき得るこのようなアッセイは、以下に簡潔に記載した蛍光活性化細胞分類(FACS)分析を利用できる。
37℃でのc−Kit mAbによる腫瘍細胞の処理後、細胞溶解物に見出される全c−Kitの量により測定して、c−Kitの分解を誘導するためのc−Kit mAbの能力を測定するためにインビトロでの細胞ベースのアッセイを使用できる。簡潔に述べると、c−Kitを発現するヒト細胞株(例えばMo7eおよびGIST882)を、3mlの無血清培地中で3×106個の細胞/mlの密度にて6ウェルディッシュ中に播種する。CK6抗体(100ng/ml)をウェルに加え、37℃にて48、24および12時間インキュベートする。細胞の1つのウェルを対照として未処理のままにする。処理時間の直後、細胞を冷PBS中で洗浄し、100μlの溶解緩衝液(50mMのHepes pH7.5、150mMのNaCl、10mMのNaPPi、50mMのNaF、1mMのNa3VO4、1%のTriton X−100、プロテアーゼ阻害剤カクテル)中で溶解させる。氷上での10分のインキュベーション後、細胞を4℃で15分間、12,000gにて遠心分離する。上清を収集し、還元条件下でウェスタンブロット分析に供する。ブロットを抗ヒトc−Kit抗体(Invitrogen,Carlsbad,CA;カタログ番号34−8800)を用いてプローブして、種々の時間のCK6とのインキュベーション前後の細胞中の全c−Kitの量を検出する。その結果は、c−Kit mAb、CK6とのインキュベーションの12時間後、未処理の細胞と比較してc−Kitの顕著な分解を示す(表9)。CK6により誘導されたc−Kitの分解は時間依存的に増加する。科学文献の以前の報告から予測されるように、37℃での12、24および48時間のSCF(100ng/ml)による腫瘍細胞の処理後、c−Kitの分解は観測されなかった(データは示さず)。
c−Kitを発現するヒト細胞株(例えばGIST882、Mo7eおよびMalm−3M)を、10%血清を使用する1つのMo7eアッセイ以外の全てについて3mlの無血清培地中の3×106個の細胞/mlの密度にて6ウェルディッシュ中に播種する。種々の濃度にてCK6抗体またはSCFをウェルに加え、37℃で24または48時間インキュベートする。細胞の1つのウェルは対照として未処理のままにする。処理時間の直後、細胞を冷PBS中で洗浄し、100μl溶解緩衝液(50mMのHepes pH7.5、150mMのNaCl、10mMのNaPPi、50mMのNaF、1mMのNa3VO4、1%のTriton−X100、プロテアーゼ阻害剤カクテル)中に溶解させる。氷上での10分のインキュベーション後、4℃で15分間、12,000gにて細胞を遠心分離する。上清を収集し、還元条件下でウェスタンブロット分析に供する。ブロットを抗ヒトc−Kit抗体(Invitrogen,Carlsbad,CA;カタログ番号34−8800またはCell Signaling Technology,Danvers,MA;カタログ番号3308)を用いてプローブして、CK6またはSCFとのインキュベーションの前後、細胞中の全c−Kitの量を検出する。その結果は、c−Kit mAb、CK6とのインキュベーション後、未処理の細胞と比較してc−Kitの顕著な分解を示さない。0%血清を使用した場合、37℃にて24時間、SCF(1ug/mlおよび100ng/ml)によるMo7e腫瘍細胞の処理後、c−Kitの分解が観測される。10%血清を使用した場合、SCFにより誘導される分解は現れない。(反復、GIST882=1、Mo7e=3、およびMalm−3M=1)。同等のタンパク質負荷を実証するために、ブロットを抗GAPDH抗体(Ambion,Carlsbad,CA;カタログ番号AM4300)を用いて再プローブする。
大部分の成体組織にわたるc−Kit発現の欠如に起因して、腫瘍内のc−Kit発現の検出は通常、c−Kit依存性細胞増殖を示唆している。ヒト癌由来の細胞株は、フローサイトメトリー分析、続いてc−Kit関連細胞ベースのリン酸化および生存アッセイを使用してc−Kit発現について試験できる。
c−KitとのSCFの相互作用は、細胞質ドメインのチロシン残基の自己リン酸化を生じる、c−Kit二量化を誘発する。c−Kitの下流で、ホスファチジルイノシトール−3−キナーゼ、Srcファミリーメンバー、JAK/STAT経路およびRas−Raf−MAPキナーゼ(MAPK)カスケードを含む、多重シグナル変換成分が活性化される。
本発明のc−Kitモノクローナル抗体による腫瘍細胞増殖の阻害は、多くの商業的に利用可能な細胞生存アッセイキットのいずれか1つを使用してインビトロアッセイにおいて試験できる。より具体的には、腫瘍細胞増殖を阻害するCK6の能力を、製造業者の指示書に従って商業的に利用可能な細胞生存キット(例えば、CellTiter−Glo(登録商標)発光細胞生存アッセイ、Promega、Madison、WI、USA)を使用して試験した。
本発明の抗c−Kit抗体の抗腫瘍効果を、当該技術分野において本質的に公知であり、広範に使用されるインビボでの異種移植片モデルに関してさらに試験できる。
腫瘍体積(mm3)=(腫瘍の長さ)(腫瘍の幅2)(π/6)
式中、腫瘍の長さは皮膚表面に平行した腫瘍の最大寸法であり、幅は皮膚表面に平行した長さに垂直な最大寸法である。体重もまた、毒性の一般的尺度として処置過程の間、頻繁に測定してもよい。T/C%およびP値はRM ANOVAにより計算し、ここで、
T/C%=[(処置体積/開始体積)/(対照体積/開始体積)]*100であり、P値は生理食塩水に関して計算する。
>CK6-CDRH1アミノ酸配列(配列番号1)
SYWIG
>CK6-CDRH2 アミノ酸配列(配列番号2)
IIYPGDSDTRYSPSFQG
>CK6-CDRH3 アミノ酸配列(配列番号3)
HGRGYNGYEGAFDI
>CK6-CDRL1 アミノ酸配列(配列番号4)
RASQGISSALA
>CK6-CDRL2 アミノ酸配列(配列番号5)
DASSLES
>CK6-CDRL3 アミノ酸配列(配列番号6)
CQQFNSYPLT
> CK6-HCVR アミノ酸配列(配列番号7)
QVQLVQSGAAVKKPGESLKISCKGSGYRFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISAGKSISTAYLQWSSLKASDTAMYYCARHGRGYNGYEGAFDIWGQGTMVTVSS
> CK6-HCVR cDNA 配列(配列番号8)
CAGGTGCAGCTGGTGCAGTCTGGAGCAGCGGTGAAAAAGCCCGGGGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGGTTTACCAGCTACTGGATCGGCTGGGTGCGCCAGATGCCCGGGAAAGGCCTGGAGTGGATGGGGATCATCTATCCTGGTGACTCTGATACCAGATACAGCCCGTCCTTCCAAGGCCAGGTCACCATCTCAGCCGGCAAGTCCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCTCGGACACCGCCATGTATTACTGTGCGAGACATGGGCGTGGATATAATGGCTACGAGGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA
> CK6-HC アミノ酸配列(配列番号9)
QVQLVQSGAAVKKPGESLKISCKGSGYRFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISAGKSISTAYLQWSSLKASDTAMYYCARHGRGYNGYEGAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
> CK6-HC cDNA 配列(配列番号10)
CAGGTGCAGCTGGTGCAGTCTGGAGCAGCGGTGAAAAAGCCCGGGGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGGTTTACCAGCTACTGGATCGGCTGGGTGCGCCAGATGCCCGGGAAAGGCCTGGAGTGGATGGGGATCATCTATCCTGGTGACTCTGATACCAGATACAGCCCGTCCTTCCAAGGCCAGGTCACCATCTCAGCCGGCAAGTCCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCTCGGACACCGCCATGTATTACTGTGCGAGACATGGGCGTGGATATAATGGCTACGAGGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA
> 分泌シグナル配列を有する完全なCK6-HC aa配列(配列番号11)
MGWSCIILFLVATATGVHSQVQLVQSGAAVKKPGESLKISCKGSGYRFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISAGKSISTAYLQWSSLKASDTAMYYCARHGRGYNGYEGAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>完全なCK6-HC cDNA配列(配列番号12)
ATGGGATGGTCATGTATCATCCTTTTTCTGGTAGCAACTGCAACTGGAGTACATTCACAGGTGCAGCTGGTGCAGTCTGGAGCAGCGGTGAAAAAGCCCGGGGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGGTTTACCAGCTACTGGATCGGCTGGGTGCGCCAGATGCCCGGGAAAGGCCTGGAGTGGATGGGGATCATCTATCCTGGTGACTCTGATACCAGATACAGCCCGTCCTTCCAAGGCCAGGTCACCATCTCAGCCGGCAAGTCCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCTCGGACACCGCCATGTATTACTGTGCGAGACATGGGCGTGGATATAATGGCTACGAGGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAATGA
> CK6-LCVR アミノ酸配列(配列番号13)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK
> CK6-LCVR cDNA 配列(配列番号14)
GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTT
> CK6-LC アミノ酸配列(配列番号15)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
> CK6-LC cDNA 配列(配列番号16)
GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG
>分泌シグナル配列を有する完全なCK6-LC aa配列(配列番号17)
MGWSCIILFLVATATGVHSAIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>完全なCK6-LC cDNA配列(配列番号18)
ATGGGATGGTCATGTATCATCCTTTTTCTGGTAGCAACTGCAACTGGAGTACATTCAGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG
>シグナルペプチドを有するヒトc-Kit 細胞外ドメイン (ECD) (配列番号19)
MRGARGAWDFLCVLLLLLRVQTGSSQPSVSPGEPSPPSIHPGKSDLIVRVGDEIRLLCTD
PGFVKWTFEILDETNENKQNEWITEKAEATNTGKYTCTNKHGLSNSIYVFVRDPAKLFLV
DRSLYGKEDNDTLVRCPLTDPEVTNYSLKGCQGKPLPKDLRFIPDPKAGIMIKSVKRAYH
RLCLHCSVDQEGKSVLSEKFILKVRPAFKAVPVVSVSKASYLLREGEEFTVTCTIKDVSS
SVYSTWKRENSQTKLQEKYNSWHHGDFNYERQATLTISSARVNDSGVFMCYANNTFGSAN
VTTTLEVVDKGFINIFPMINTTVFVNDGENVDLIVEYEAFPKPEHQQWIYMNRTFTDKWE
DYPKSENESNIRYVSELHLTRLKGTEGGTYTFLVSNSDVNAAIAFNVYVNTKPEILTYDR
LVNGMLQCVAAGFPEPTIDWYFCPGTEQRCSASVLPVDVQTLNSSGPPFGKLVVQSSIDS
SAFKHNGTVECKAYNDVGKTSAYFNFAFKGNNKEQIHPHT
>シグナルペプチドを有さないヒトc-Kit細胞外ドメイン(ECD) (配列番号20)
QPSVSPGEPSPPSIHPGKSDLIVRVGDEIRLLCTDPGFVKWTFEILDETNENKQNEWITEKAEATNT
GKYTCTNKHGLSNSIYVFVRDPAKLFLVDRSLYGKEDNDTLVRCPLTDPEVTNYSLKGCQGKPLPKD
LRFIPDPKAGIMIKSVKRAYHRLCLHCSVDQEGKSVLSEKFILKVRPAFKAVPVVSVSKASYLLREG
EEFTVTCTIKDVSSSVYSTWKRENSQTKLQEKYNSWHHGDFNYERQATLTISSARVNDSGVFMCYAN
NTFGSANVTTTLEVVDKGFINIFPMINTTVFVNDGENVDLIVEYEAFPKPEHQQWIYMNRTFTDKWE
DYPKSENESNIRYVSELHLTRLKGTEGGTYTFLVSNSDVNAAIAFNVYVNTKPEILTYDRLVNGMLQ
CVAAGFPEPTIDWYFCPGTEQRCSASVLPVDVQTLNSSGPPFGKLVVQSSIDSSAFKHNGTVECKAY
NDVGKTSAYFNFAFKGNNKEQIHPHT
Claims (5)
- 配列番号20のアミノ酸配列からなるヒトc−Kitの細胞外ドメイン(ECD)に特異的に結合する抗体であって、前記抗体は重鎖可変領域(HCVR)および軽鎖可変領域(LCVR)を含み、前記HCVRはCDRH1、CDRH2およびCDRH3アミノ酸配列を含み、前記LCVRはCDRL1、CDRL2およびCDRL3アミノ酸配列を含み、CDRH1はSYWIG(配列番号1)であり、CDRH2はIIYPGDSDTRYSPSFQG(配列番号2)であり、CDRH3はHGRGYNGYEGAFDI(配列番号3)であり、CDRL1はRASQGISSALA(配列番号4)であり、CDRL2はDASSLES(配列番号5)であり、CDRL3はCQQFNSYPLT(配列番号6)である、抗体。
- 前記LCVRアミノ酸配列は配列番号13であり、前記HCVRアミノ酸配列は配列番号7である、請求項1に記載の抗体。
- 重鎖および軽鎖を含み、前記重鎖アミノ酸配列は配列番号9であり、前記軽鎖アミノ酸配列は配列番号15である、請求項1または2に記載の抗体。
- 2本の重鎖および2本の軽鎖を含み、前記重鎖アミノ酸配列の各々は配列番号9であり、前記軽鎖アミノ酸配列の各々は配列番号15である、請求項1〜3のいずれか一項に記載の抗体。
- 請求項1〜4のいずれか一項に記載の抗体の抗原結合断片。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201161485255P | 2011-05-12 | 2011-05-12 | |
US61/485,255 | 2011-05-12 | ||
PCT/US2012/036240 WO2012154480A1 (en) | 2011-05-12 | 2012-05-03 | c-KIT ANTIBODIES AND USES THEREOF |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2014515030A JP2014515030A (ja) | 2014-06-26 |
JP5899310B2 true JP5899310B2 (ja) | 2016-04-06 |
Family
ID=46046358
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2014510362A Active JP5899310B2 (ja) | 2011-05-12 | 2012-05-03 | c−KIT抗体およびその使用 |
Country Status (13)
Country | Link |
---|---|
US (1) | US8552157B2 (ja) |
EP (1) | EP2707028B1 (ja) |
JP (1) | JP5899310B2 (ja) |
KR (1) | KR101601387B1 (ja) |
CN (1) | CN103533960B (ja) |
AR (1) | AR086044A1 (ja) |
AU (1) | AU2012253943B2 (ja) |
BR (1) | BR112013028908A2 (ja) |
CA (1) | CA2835200C (ja) |
EA (1) | EA023665B1 (ja) |
MX (1) | MX337577B (ja) |
TW (1) | TWI532748B (ja) |
WO (1) | WO2012154480A1 (ja) |
Families Citing this family (75)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DK2668210T3 (da) | 2011-01-26 | 2020-08-24 | Celldex Therapeutics Inc | Anti-kit antistoffer og anvendelser deraf |
CN116574185A (zh) | 2012-07-25 | 2023-08-11 | 塞尔德克斯医疗公司 | 抗kit抗体及其用途 |
NZ710929A (en) * | 2013-03-15 | 2018-02-23 | Novartis Ag | Antibody drug conjugates |
WO2015050959A1 (en) * | 2013-10-01 | 2015-04-09 | Yale University | Anti-kit antibodies and methods of use thereof |
EP3065773A1 (en) * | 2013-11-05 | 2016-09-14 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Human neutralizing anti-kit antibodies and uses thereof |
EP3593812A3 (en) | 2014-03-15 | 2020-05-27 | Novartis AG | Treatment of cancer using chimeric antigen receptor |
CN113908269A (zh) * | 2014-05-23 | 2022-01-11 | 塞尔德克斯医疗公司 | 嗜酸性粒细胞或肥大细胞相关病症的治疗 |
JP2017528433A (ja) | 2014-07-21 | 2017-09-28 | ノバルティス アーゲー | 低い免疫増強用量のmTOR阻害剤とCARの組み合わせ |
US20180334490A1 (en) | 2014-12-03 | 2018-11-22 | Qilong H. Wu | Methods for b cell preconditioning in car therapy |
WO2016126608A1 (en) | 2015-02-02 | 2016-08-11 | Novartis Ag | Car-expressing cells against multiple tumor antigens and uses thereof |
EP3280720A4 (en) | 2015-04-06 | 2018-12-05 | President and Fellows of Harvard College | Compositions and methods for non-myeloablative conditioning |
EP3286211A1 (en) | 2015-04-23 | 2018-02-28 | Novartis AG | Treatment of cancer using chimeric antigen receptor and protein kinase a blocker |
US11667691B2 (en) | 2015-08-07 | 2023-06-06 | Novartis Ag | Treatment of cancer using chimeric CD3 receptor proteins |
EP3380620A1 (en) | 2015-11-23 | 2018-10-03 | Novartis AG | Optimized lentiviral transfer vectors and uses thereof |
ES2944597T3 (es) | 2015-12-30 | 2023-06-22 | Novartis Ag | Terapias con células efectoras inmunitarias de eficacia mejorada |
WO2017149515A1 (en) | 2016-03-04 | 2017-09-08 | Novartis Ag | Cells expressing multiple chimeric antigen receptor (car) molecules and uses therefore |
US11549099B2 (en) | 2016-03-23 | 2023-01-10 | Novartis Ag | Cell secreted minibodies and uses thereof |
SG11201809041TA (en) | 2016-04-15 | 2018-11-29 | Novartis Ag | Compositions and methods for selective protein expression |
US10111966B2 (en) | 2016-06-17 | 2018-10-30 | Magenta Therapeutics, Inc. | Methods for the depletion of CD117+ cells |
EP3490590A2 (en) | 2016-08-01 | 2019-06-05 | Novartis AG | Treatment of cancer using a chimeric antigen receptor in combination with an inhibitor of a pro-m2 macrophage molecule |
WO2018111340A1 (en) | 2016-12-16 | 2018-06-21 | Novartis Ag | Methods for determining potency and proliferative function of chimeric antigen receptor (car)-t cells |
IL268058B2 (en) | 2017-01-20 | 2023-09-01 | Magenta Therapeutics Inc | Compositions and methods for depleting cd137 plus cells |
US11535662B2 (en) | 2017-01-26 | 2022-12-27 | Novartis Ag | CD28 compositions and methods for chimeric antigen receptor therapy |
US20190375815A1 (en) | 2017-01-31 | 2019-12-12 | Novartis Ag | Treatment of cancer using chimeric t cell receptor proteins having multiple specificities |
EP3589647A1 (en) | 2017-02-28 | 2020-01-08 | Novartis AG | Shp inhibitor compositions and uses for chimeric antigen receptor therapy |
WO2018229715A1 (en) | 2017-06-16 | 2018-12-20 | Novartis Ag | Compositions comprising anti-cd32b antibodies and methods of use thereof |
CN111629749A (zh) | 2017-10-18 | 2020-09-04 | 诺华股份有限公司 | 用于选择性蛋白质降解的组合物和方法 |
WO2019084057A2 (en) | 2017-10-24 | 2019-05-02 | Magenta Therapeutics, Inc. | COMPOSITIONS AND METHODS FOR DEPLOYING CD117 + CELLS |
WO2019084053A1 (en) * | 2017-10-24 | 2019-05-02 | Magenta Therapeutics, Inc. | COMPOSITIONS AND METHODS FOR DEPLOYING CD117 + CELLS |
US10899843B2 (en) * | 2017-10-24 | 2021-01-26 | Magenta Therapeutics, Inc. | Compositions and methods for the depletion of CD117+ cells |
KR20200069358A (ko) | 2017-10-25 | 2020-06-16 | 노파르티스 아게 | 키메라 항원 수용체 발현 세포의 제조 방법 |
EP3700933A1 (en) | 2017-10-25 | 2020-09-02 | Novartis AG | Antibodies targeting cd32b and methods of use thereof |
US20210179709A1 (en) | 2017-10-31 | 2021-06-17 | Novartis Ag | Anti-car compositions and methods |
US20210047405A1 (en) | 2018-04-27 | 2021-02-18 | Novartis Ag | Car t cell therapies with enhanced efficacy |
WO2019213282A1 (en) | 2018-05-01 | 2019-11-07 | Novartis Ag | Biomarkers for evaluating car-t cells to predict clinical outcome |
EP3801769A1 (en) | 2018-05-25 | 2021-04-14 | Novartis AG | Combination therapy with chimeric antigen receptor (car) therapies |
CA3101943A1 (en) * | 2018-06-07 | 2019-12-12 | Magenta Therapeutics, Inc. | Therapeutic methods using antibody drug conjugates (adcs) |
CN108822212B (zh) * | 2018-06-08 | 2021-06-29 | 北京大学 | 选择性识别致癌性突变体kit itd的单克隆抗体及其应用 |
EP3802825A1 (en) | 2018-06-08 | 2021-04-14 | Intellia Therapeutics, Inc. | Compositions and methods for immunooncology |
AR116109A1 (es) | 2018-07-10 | 2021-03-31 | Novartis Ag | Derivados de 3-(5-amino-1-oxoisoindolin-2-il)piperidina-2,6-diona y usos de los mismos |
KR20200040407A (ko) * | 2018-10-10 | 2020-04-20 | 주식회사 노벨티노빌리티 | 신규 항-c-KIT 항체 |
KR20210094610A (ko) * | 2018-11-26 | 2021-07-29 | 포티 세븐, 인코포레이티드 | c-Kit에 대한 인간화 항체 |
AU2019402189B2 (en) | 2018-12-20 | 2023-04-13 | Novartis Ag | Dosing regimen and pharmaceutical combination comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives |
BR112021015672A2 (pt) | 2019-02-15 | 2021-10-05 | Novartis Ag | Derivados de 3-(1-oxoisoindolin-2-il)piperidina-2,6-diona substituída e usos dos mesmos |
BR112021015783A2 (pt) | 2019-02-15 | 2021-10-05 | Novartis Ag | Derivados de 3-(1-oxo-5-(piperidin-4-il)isoindolin-2-il)piperidina-2,6-diona e usos dos mesmos |
WO2020219742A1 (en) | 2019-04-24 | 2020-10-29 | Novartis Ag | Compositions and methods for selective protein degradation |
EP3959242A4 (en) * | 2019-04-24 | 2023-09-13 | Magenta Therapeutics, Inc. | ANTRACYCLINE ANTIBODY-DRUG CONJUGATES AND THEIR USES |
JP2022529727A (ja) * | 2019-04-24 | 2022-06-23 | マジェンタ セラピューティクス インコーポレイテッド | Anti-cd117 とその使用 |
WO2020219770A1 (en) * | 2019-04-24 | 2020-10-29 | Magenta Therapeutics, Inc. | Anti-cd117 antibodies and uses thereof |
EP3825330A1 (en) | 2019-11-19 | 2021-05-26 | International-Drug-Development-Biotech | Anti-cd117 antibodies and methods of use thereof |
KR20210063782A (ko) * | 2019-11-25 | 2021-06-02 | 주식회사 노벨티노빌리티 | c-kit에 대한 항체 및 이의 용도 |
KR20220116257A (ko) | 2019-12-20 | 2022-08-22 | 노파르티스 아게 | 골수섬유증 및 골수이형성 증후군을 치료하기 위한, 데시타빈 또는 항 pd-1 항체 스파르탈리주맙을 포함하거나 또는 포함하지 않는, 항 tim-3 항체 mbg453 및 항 tgf-베타 항체 nis793의 조합물 |
MX2022007930A (es) | 2019-12-24 | 2022-08-08 | Carna Biosciences Inc | Compuestos moduladores de diacilglicerol quinasa. |
WO2021163064A2 (en) | 2020-02-14 | 2021-08-19 | Jounce Therapeutics, Inc. | Antibodies and fusion proteins that bind to ccr8 and uses thereof |
CA3185455A1 (en) | 2020-06-11 | 2021-12-16 | Novartis Ag | Zbtb32 inhibitors and uses thereof |
CN115916199A (zh) | 2020-06-23 | 2023-04-04 | 诺华股份有限公司 | 包含3-(1-氧代异吲哚啉-2-基)哌啶-2,6-二酮衍生物的给药方案 |
JP2023536164A (ja) | 2020-08-03 | 2023-08-23 | ノバルティス アーゲー | ヘテロアリール置換3-(1-オキソイソインドリン-2-イル)ピペリジン-2,6-ジオン誘導体及びその使用 |
TW202304979A (zh) | 2021-04-07 | 2023-02-01 | 瑞士商諾華公司 | 抗TGFβ抗體及其他治療劑用於治療增殖性疾病之用途 |
TW202309294A (zh) | 2021-04-27 | 2023-03-01 | 瑞士商諾華公司 | 病毒載體生產系統 |
AU2022299051A1 (en) | 2021-06-23 | 2023-12-07 | Gilead Sciences, Inc. | Diacylglyercol kinase modulating compounds |
CA3222439A1 (en) | 2021-06-23 | 2022-12-29 | Gilead Sciences, Inc. | Diacylglyercol kinase modulating compounds |
EP4359413A1 (en) | 2021-06-23 | 2024-05-01 | Gilead Sciences, Inc. | Diacylglyercol kinase modulating compounds |
WO2022271677A1 (en) | 2021-06-23 | 2022-12-29 | Gilead Sciences, Inc. | Diacylglyercol kinase modulating compounds |
AU2022375782A1 (en) | 2021-10-28 | 2024-05-02 | Gilead Sciences, Inc. | Pyridizin-3(2h)-one derivatives |
AU2022376954A1 (en) | 2021-10-29 | 2024-05-02 | Gilead Sciences, Inc. | Cd73 compounds |
WO2023122581A2 (en) | 2021-12-22 | 2023-06-29 | Gilead Sciences, Inc. | Ikaros zinc finger family degraders and uses thereof |
AU2022417491A1 (en) | 2021-12-22 | 2024-05-23 | Gilead Sciences, Inc. | Ikaros zinc finger family degraders and uses thereof |
KR102544135B1 (ko) * | 2022-01-26 | 2023-06-19 | 주식회사 노벨티노빌리티 | c-Kit을 표적으로 하는 면역접합체 |
TW202340168A (zh) | 2022-01-28 | 2023-10-16 | 美商基利科學股份有限公司 | Parp7抑制劑 |
WO2023178181A1 (en) | 2022-03-17 | 2023-09-21 | Gilead Sciences, Inc. | Ikaros zinc finger family degraders and uses thereof |
TW202400138A (zh) | 2022-04-21 | 2024-01-01 | 美商基利科學股份有限公司 | Kras g12d調節化合物 |
WO2023214325A1 (en) | 2022-05-05 | 2023-11-09 | Novartis Ag | Pyrazolopyrimidine derivatives and uses thereof as tet2 inhibitors |
WO2024006929A1 (en) | 2022-07-01 | 2024-01-04 | Gilead Sciences, Inc. | Cd73 compounds |
WO2024040195A1 (en) | 2022-08-17 | 2024-02-22 | Capstan Therapeutics, Inc. | Conditioning for in vivo immune cell engineering |
WO2024089639A1 (en) | 2022-10-26 | 2024-05-02 | Novartis Ag | Lentiviral formulations |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE4205148A1 (de) * | 1991-05-25 | 1993-01-21 | Boehringer Mannheim Gmbh | Monoklonale antikoerper gegen c-kit |
GB0216081D0 (en) | 2002-07-11 | 2002-08-21 | Imp College Innovations Ltd | Methods of making biological materials and uses thereof |
TWI395754B (zh) * | 2006-04-24 | 2013-05-11 | Amgen Inc | 人類化之c-kit抗體 |
CN101801407B (zh) | 2007-06-05 | 2013-12-18 | 耶鲁大学 | 受体酪氨酸激酶抑制剂及其使用方法 |
JO3096B1 (ar) | 2008-11-07 | 2017-03-15 | Imclone Llc | الأجسام المضادة لمستقبل ii مضاد tgfb |
-
2012
- 2012-04-24 AR ARP120101413A patent/AR086044A1/es unknown
- 2012-04-27 TW TW101115271A patent/TWI532748B/zh not_active IP Right Cessation
- 2012-05-03 JP JP2014510362A patent/JP5899310B2/ja active Active
- 2012-05-03 MX MX2013013226A patent/MX337577B/es active IP Right Grant
- 2012-05-03 KR KR1020137029568A patent/KR101601387B1/ko not_active IP Right Cessation
- 2012-05-03 AU AU2012253943A patent/AU2012253943B2/en not_active Ceased
- 2012-05-03 CN CN201280022747.6A patent/CN103533960B/zh not_active Expired - Fee Related
- 2012-05-03 EA EA201391507A patent/EA023665B1/ru not_active IP Right Cessation
- 2012-05-03 BR BR112013028908A patent/BR112013028908A2/pt not_active IP Right Cessation
- 2012-05-03 EP EP12719889.3A patent/EP2707028B1/en not_active Not-in-force
- 2012-05-03 US US13/462,853 patent/US8552157B2/en active Active
- 2012-05-03 CA CA2835200A patent/CA2835200C/en not_active Expired - Fee Related
- 2012-05-03 WO PCT/US2012/036240 patent/WO2012154480A1/en active Application Filing
Also Published As
Publication number | Publication date |
---|---|
AR086044A1 (es) | 2013-11-13 |
TWI532748B (zh) | 2016-05-11 |
EA201391507A1 (ru) | 2014-03-31 |
US8552157B2 (en) | 2013-10-08 |
US20120288506A1 (en) | 2012-11-15 |
EP2707028B1 (en) | 2017-03-22 |
TW201247707A (en) | 2012-12-01 |
WO2012154480A1 (en) | 2012-11-15 |
BR112013028908A2 (pt) | 2017-01-31 |
CN103533960B (zh) | 2015-08-12 |
MX2013013226A (es) | 2013-12-12 |
CN103533960A (zh) | 2014-01-22 |
MX337577B (es) | 2016-03-10 |
EA023665B1 (ru) | 2016-06-30 |
KR101601387B1 (ko) | 2016-03-08 |
CA2835200C (en) | 2017-04-11 |
EP2707028A1 (en) | 2014-03-19 |
AU2012253943A1 (en) | 2013-10-24 |
JP2014515030A (ja) | 2014-06-26 |
KR20140027239A (ko) | 2014-03-06 |
CA2835200A1 (en) | 2012-11-15 |
AU2012253943B2 (en) | 2016-12-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5899310B2 (ja) | c−KIT抗体およびその使用 | |
JP7257364B2 (ja) | 抗cd137抗体 | |
TWI381848B (zh) | 抗纖維母細胞生長因子受體-3 (fgfr-3) 抗體及包含其之醫藥組合物 | |
US20090246205A1 (en) | Inhibition of macrophage-stimulating protein receptor (ron) | |
CN105829345B (zh) | 结合egfr和met的多功能抗体 | |
JP2011520885A (ja) | 抗cxcr4抗体 | |
MX2012011234A (es) | Anticuerpos contra csf-1r. | |
US20220411501A1 (en) | Anti-cd117 antibodies and methods of use thereof | |
EP3145544B1 (en) | Ang2 antibodies | |
JP2023502023A (ja) | Cd200受容体アンタゴニスト結合分子 | |
TW201716439A (zh) | Her3抗體 | |
AU2015264552A1 (en) | VEGFR2/Ang2 compounds | |
JP2018505144A (ja) | 抗cxcl12抗体分子およびその使用 | |
WO2024102962A1 (en) | Cytotoxic bispecific antibodies binding to dr5 and muc16 and uses thereof | |
KR20230034367A (ko) | 항-종양 괴사 인자 수용체 (tnfr2) 항체 및 그 용도 | |
CN116262788A (zh) | 抗cd27抗体分子 | |
EP3440111A1 (en) | Anti-vegfr-1 antibodies and uses thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20140509 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20150630 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20160209 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20160307 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5899310 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |