JP2022176381A - Antibody-drug conjugates for treating breast cancer or gastric cancer - Google Patents
Antibody-drug conjugates for treating breast cancer or gastric cancer Download PDFInfo
- Publication number
- JP2022176381A JP2022176381A JP2022162603A JP2022162603A JP2022176381A JP 2022176381 A JP2022176381 A JP 2022176381A JP 2022162603 A JP2022162603 A JP 2022162603A JP 2022162603 A JP2022162603 A JP 2022162603A JP 2022176381 A JP2022176381 A JP 2022176381A
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- drug
- sugar chain
- adc
- her2
- 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.)
- Pending
Links
- 229940049595 antibody-drug conjugate Drugs 0.000 title claims abstract description 90
- 239000000611 antibody drug conjugate Substances 0.000 title claims abstract description 88
- 206010006187 Breast cancer Diseases 0.000 title claims abstract description 31
- 208000026310 Breast neoplasm Diseases 0.000 title claims abstract description 31
- 208000005718 Stomach Neoplasms Diseases 0.000 title claims abstract description 30
- 206010017758 gastric cancer Diseases 0.000 title claims abstract description 30
- 201000011549 stomach cancer Diseases 0.000 title claims abstract description 30
- 235000000346 sugar Nutrition 0.000 claims abstract description 85
- 239000003814 drug Substances 0.000 claims abstract description 54
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 19
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 9
- 102000005600 Cathepsins Human genes 0.000 claims abstract description 8
- 108010084457 Cathepsins Proteins 0.000 claims abstract description 8
- 229940124597 therapeutic agent Drugs 0.000 claims abstract description 6
- 229960002173 citrulline Drugs 0.000 claims abstract description 5
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 5
- 229960000575 trastuzumab Drugs 0.000 claims description 40
- 108010093470 monomethyl auristatin E Proteins 0.000 claims description 35
- 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 claims description 34
- 108010044540 auristatin Proteins 0.000 claims description 8
- 108010059074 monomethylauristatin F Proteins 0.000 claims description 7
- MFRNYXJJRJQHNW-DEMKXPNLSA-N (2s)-2-[[(2r,3r)-3-methoxy-3-[(2s)-1-[(3r,4s,5s)-3-methoxy-5-methyl-4-[methyl-[(2s)-3-methyl-2-[[(2s)-3-methyl-2-(methylamino)butanoyl]amino]butanoyl]amino]heptanoyl]pyrrolidin-2-yl]-2-methylpropanoyl]amino]-3-phenylpropanoic acid Chemical compound CN[C@@H](C(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(O)=O)CC1=CC=CC=C1 MFRNYXJJRJQHNW-DEMKXPNLSA-N 0.000 claims description 6
- BLUGYPPOFIHFJS-UUFHNPECSA-N (2s)-n-[(2s)-1-[[(3r,4s,5s)-3-methoxy-1-[(2s)-2-[(1r,2r)-1-methoxy-2-methyl-3-oxo-3-[[(1s)-2-phenyl-1-(1,3-thiazol-2-yl)ethyl]amino]propyl]pyrrolidin-1-yl]-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]-3-methyl-2-(methylamino)butanamid Chemical compound CN[C@@H](C(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=1SC=CN=1)CC1=CC=CC=C1 BLUGYPPOFIHFJS-UUFHNPECSA-N 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 description 53
- 229940079593 drug Drugs 0.000 description 48
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 35
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 35
- 230000000694 effects Effects 0.000 description 19
- 238000006243 chemical reaction Methods 0.000 description 14
- 230000027455 binding Effects 0.000 description 13
- 238000009739 binding Methods 0.000 description 13
- 238000000034 method Methods 0.000 description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 206010028980 Neoplasm Diseases 0.000 description 12
- 239000008363 phosphate buffer Substances 0.000 description 12
- 239000000243 solution Substances 0.000 description 11
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 10
- 230000015572 biosynthetic process Effects 0.000 description 10
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- 201000011510 cancer Diseases 0.000 description 9
- 230000022534 cell killing Effects 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 238000003786 synthesis reaction Methods 0.000 description 8
- -1 thioteva Chemical compound 0.000 description 8
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 7
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 7
- 125000000524 functional group Chemical group 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- OVRNDRQMDRJTHS-RTRLPJTCSA-N N-acetyl-D-glucosamine Chemical group CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-RTRLPJTCSA-N 0.000 description 6
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 6
- 230000001093 anti-cancer Effects 0.000 description 6
- 229950006780 n-acetylglucosamine Drugs 0.000 description 6
- 238000004587 chromatography analysis Methods 0.000 description 5
- 235000018417 cysteine Nutrition 0.000 description 5
- 230000001472 cytotoxic effect Effects 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 229920001223 polyethylene glycol Polymers 0.000 description 5
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 4
- 102000001301 EGF receptor Human genes 0.000 description 4
- 108060006698 EGF receptor Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 4
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 231100000433 cytotoxic Toxicity 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- IMSODMZESSGVBE-UHFFFAOYSA-N 2-Oxazoline Chemical compound C1CN=CO1 IMSODMZESSGVBE-UHFFFAOYSA-N 0.000 description 3
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 3
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 3
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 3
- CDOJPCSDOXYJJF-CBTAGEKQSA-N N,N'-diacetylchitobiose Chemical group O[C@@H]1[C@@H](NC(=O)C)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CDOJPCSDOXYJJF-CBTAGEKQSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 102000004243 Tubulin Human genes 0.000 description 3
- 108090000704 Tubulin Proteins 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- 229940124651 anti-breast cancer agent Drugs 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 description 3
- 239000003183 carcinogenic agent Substances 0.000 description 3
- 238000012650 click reaction Methods 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 125000000151 cysteine group Chemical class N[C@@H](CS)C(=O)* 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000006206 glycosylation reaction Methods 0.000 description 3
- 235000018977 lysine Nutrition 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000000600 sorbitol Substances 0.000 description 3
- 235000010356 sorbitol Nutrition 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 125000003504 2-oxazolinyl group Chemical group O1C(=NCC1)* 0.000 description 2
- 208000007934 ACTH-independent macronodular adrenal hyperplasia Diseases 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 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 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 2
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 2
- 102000051366 Glycosyltransferases Human genes 0.000 description 2
- 108700023372 Glycosyltransferases Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- DUKURNFHYQXCJG-UHFFFAOYSA-N Lewis A pentasaccharide Natural products OC1C(O)C(O)C(C)OC1OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(NC(C)=O)C(OC2C(C(OC3C(OC(O)C(O)C3O)CO)OC(CO)C2O)O)OC1CO DUKURNFHYQXCJG-UHFFFAOYSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 2
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 2
- 108060008539 Transglutaminase Proteins 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 2
- 208000009956 adenocarcinoma Diseases 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- 150000001345 alkine derivatives Chemical class 0.000 description 2
- 229940100198 alkylating agent Drugs 0.000 description 2
- 239000002168 alkylating agent Substances 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000340 anti-metabolite Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 229940125644 antibody drug Drugs 0.000 description 2
- 229940100197 antimetabolite Drugs 0.000 description 2
- 239000002256 antimetabolite Substances 0.000 description 2
- 229940034982 antineoplastic agent Drugs 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 238000006664 bond formation reaction Methods 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 229960000684 cytarabine Drugs 0.000 description 2
- 238000002784 cytotoxicity assay Methods 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 229960002949 fluorouracil Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 150000008267 fucoses Chemical class 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 239000012216 imaging agent Substances 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000003588 lysine group Chemical class [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- DASWEROEPLKSEI-UIJRFTGLSA-N monomethyl auristatin e Chemical compound CN[C@@H](C(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 DASWEROEPLKSEI-UIJRFTGLSA-N 0.000 description 2
- 229940127084 other anti-cancer agent Drugs 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000002953 preparative HPLC Methods 0.000 description 2
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 229960001674 tegafur Drugs 0.000 description 2
- WFWLQNSHRPWKFK-ZCFIWIBFSA-N tegafur Chemical compound O=C1NC(=O)C(F)=CN1[C@@H]1OCCC1 WFWLQNSHRPWKFK-ZCFIWIBFSA-N 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 2
- 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 2
- 102000003601 transglutaminase Human genes 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- WDQLRUYAYXDIFW-RWKIJVEZSA-N (2r,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-4-[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-6-[[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](CO)O[C@@H](O)[C@H](O)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1 WDQLRUYAYXDIFW-RWKIJVEZSA-N 0.000 description 1
- AGGWFDNPHKLBBV-YUMQZZPRSA-N (2s)-2-[[(2s)-2-amino-3-methylbutanoyl]amino]-5-(carbamoylamino)pentanoic acid Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(O)=O)CCCNC(N)=O AGGWFDNPHKLBBV-YUMQZZPRSA-N 0.000 description 1
- FPVKHBSQESCIEP-UHFFFAOYSA-N (8S)-3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,6,7,8-tetrahydroimidazo[4,5-d][1,3]diazepin-8-ol Natural products C1C(O)C(CO)OC1N1C(NC=NCC2O)=C2N=C1 FPVKHBSQESCIEP-UHFFFAOYSA-N 0.000 description 1
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 1
- LKJPYSCBVHEWIU-KRWDZBQOSA-N (R)-bicalutamide Chemical compound C([C@@](O)(C)C(=O)NC=1C=C(C(C#N)=CC=1)C(F)(F)F)S(=O)(=O)C1=CC=C(F)C=C1 LKJPYSCBVHEWIU-KRWDZBQOSA-N 0.000 description 1
- 102100025573 1-alkyl-2-acetylglycerophosphocholine esterase Human genes 0.000 description 1
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- FPVCVHVTMPCZTH-UHFFFAOYSA-N 2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]ethanamine Chemical compound NCCOCCOCCOCCN=[N+]=[N-] FPVCVHVTMPCZTH-UHFFFAOYSA-N 0.000 description 1
- SVZIASXLJNEQGJ-UHFFFAOYSA-N 2-azido-4,5-dihydro-1,3-oxazole Chemical compound N(=[N+]=[N-])C=1OCCN=1 SVZIASXLJNEQGJ-UHFFFAOYSA-N 0.000 description 1
- JNUTUYIONKSVDZ-UHFFFAOYSA-M 2-chloro-1,3-dimethylbenzimidazol-3-ium;chloride Chemical compound [Cl-].C1=CC=C2N(C)C(Cl)=[N+](C)C2=C1 JNUTUYIONKSVDZ-UHFFFAOYSA-M 0.000 description 1
- UZFPOOOQHWICKY-UHFFFAOYSA-N 3-[13-[1-[1-[8,12-bis(2-carboxyethyl)-17-(1-hydroxyethyl)-3,7,13,18-tetramethyl-21,24-dihydroporphyrin-2-yl]ethoxy]ethyl]-18-(2-carboxyethyl)-8-(1-hydroxyethyl)-3,7,12,17-tetramethyl-22,23-dihydroporphyrin-2-yl]propanoic acid Chemical compound N1C(C=C2C(=C(CCC(O)=O)C(C=C3C(=C(C)C(C=C4N5)=N3)CCC(O)=O)=N2)C)=C(C)C(C(C)O)=C1C=C5C(C)=C4C(C)OC(C)C1=C(N2)C=C(N3)C(C)=C(C(O)C)C3=CC(C(C)=C3CCC(O)=O)=NC3=CC(C(CCC(O)=O)=C3C)=NC3=CC2=C1C UZFPOOOQHWICKY-UHFFFAOYSA-N 0.000 description 1
- 125000004042 4-aminobutyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])N([H])[H] 0.000 description 1
- VVIAGPKUTFNRDU-UHFFFAOYSA-N 6S-folinic acid Natural products C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-UHFFFAOYSA-N 0.000 description 1
- KABRXLINDSPGDF-UHFFFAOYSA-N 7-bromoisoquinoline Chemical compound C1=CN=CC2=CC(Br)=CC=C21 KABRXLINDSPGDF-UHFFFAOYSA-N 0.000 description 1
- NKGPJODWTZCHGF-UHFFFAOYSA-N 9-[3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-3h-purine-6-thione Chemical compound OC1C(O)C(CO)OC1N1C(NC=NC2=S)=C2N=C1 NKGPJODWTZCHGF-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 108010055851 Acetylglucosaminidase Proteins 0.000 description 1
- 239000012114 Alexa Fluor 647 Substances 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 206010002199 Anaphylactic shock Diseases 0.000 description 1
- 108010024976 Asparaginase Proteins 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- VGGGPCQERPFHOB-MCIONIFRSA-N Bestatin Chemical compound CC(C)C[C@H](C(O)=O)NC(=O)[C@@H](O)[C@H](N)CC1=CC=CC=C1 VGGGPCQERPFHOB-MCIONIFRSA-N 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- AOCCBINRVIKJHY-UHFFFAOYSA-N Carmofur Chemical compound CCCCCCNC(=O)N1C=C(F)C(=O)NC1=O AOCCBINRVIKJHY-UHFFFAOYSA-N 0.000 description 1
- JWBOIMRXGHLCPP-UHFFFAOYSA-N Chloditan Chemical compound C=1C=CC=C(Cl)C=1C(C(Cl)Cl)C1=CC=C(Cl)C=C1 JWBOIMRXGHLCPP-UHFFFAOYSA-N 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 108010005843 Cysteine Proteases Proteins 0.000 description 1
- 102000005927 Cysteine Proteases Human genes 0.000 description 1
- 108010092160 Dactinomycin Proteins 0.000 description 1
- 238000005698 Diels-Alder reaction Methods 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
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- SAMRUMKYXPVKPA-VFKOLLTISA-N Enocitabine Chemical compound O=C1N=C(NC(=O)CCCCCCCCCCCCCCCCCCCCC)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 SAMRUMKYXPVKPA-VFKOLLTISA-N 0.000 description 1
- OBMLHUPNRURLOK-XGRAFVIBSA-N Epitiostanol Chemical compound C1[C@@H]2S[C@@H]2C[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 OBMLHUPNRURLOK-XGRAFVIBSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 235000016623 Fragaria vesca Nutrition 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 1
- 102000030902 Galactosyltransferase Human genes 0.000 description 1
- 108060003306 Galactosyltransferase Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 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
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108010069236 Goserelin Proteins 0.000 description 1
- VSNHCAURESNICA-UHFFFAOYSA-N Hydroxyurea Chemical compound NC(=O)NO VSNHCAURESNICA-UHFFFAOYSA-N 0.000 description 1
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical group CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 1
- 244000246386 Mentha pulegium Species 0.000 description 1
- 235000016257 Mentha pulegium Nutrition 0.000 description 1
- 235000004357 Mentha x piperita Nutrition 0.000 description 1
- IVDYZAAPOLNZKG-KWHRADDSSA-N Mepitiostane Chemical compound O([C@@H]1[C@]2(CC[C@@H]3[C@@]4(C)C[C@H]5S[C@H]5C[C@@H]4CC[C@H]3[C@@H]2CC1)C)C1(OC)CCCC1 IVDYZAAPOLNZKG-KWHRADDSSA-N 0.000 description 1
- 238000006845 Michael addition reaction Methods 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000872931 Myoporum sandwicense Species 0.000 description 1
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 1
- 101710204212 Neocarzinostatin Proteins 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 108010057150 Peplomycin Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102100030122 Protein O-GlcNAcase Human genes 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 241000508269 Psidium Species 0.000 description 1
- AHHFEZNOXOZZQA-ZEBDFXRSSA-N Ranimustine Chemical compound CO[C@H]1O[C@H](CNC(=O)N(CCCl)N=O)[C@@H](O)[C@H](O)[C@H]1O AHHFEZNOXOZZQA-ZEBDFXRSSA-N 0.000 description 1
- 229920002305 Schizophyllan Polymers 0.000 description 1
- 102000003838 Sialyltransferases Human genes 0.000 description 1
- 108090000141 Sialyltransferases Proteins 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- OCOKWVBYZHBHLU-UHFFFAOYSA-N Sobuzoxane Chemical compound C1C(=O)N(COC(=O)OCC(C)C)C(=O)CN1CCN1CC(=O)N(COC(=O)OCC(C)C)C(=O)C1 OCOKWVBYZHBHLU-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 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 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- KBGAYAKRZNYFFG-BOHATCBPSA-N aceneuramic acid Chemical group OC(=O)C(=O)C[C@H](O)[C@@H](NC(=O)C)[C@@H](O)[C@H](O)[C@H](O)CO KBGAYAKRZNYFFG-BOHATCBPSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 238000010976 amide bond formation reaction Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- YBBLVLTVTVSKRW-UHFFFAOYSA-N anastrozole Chemical compound N#CC(C)(C)C1=CC(C(C)(C#N)C)=CC(CN2N=CN=C2)=C1 YBBLVLTVTVSKRW-UHFFFAOYSA-N 0.000 description 1
- 229960002932 anastrozole Drugs 0.000 description 1
- 239000002839 antibiotic anticancer agent Substances 0.000 description 1
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- KLNFSAOEKUDMFA-UHFFFAOYSA-N azanide;2-hydroxyacetic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OCC(O)=O KLNFSAOEKUDMFA-UHFFFAOYSA-N 0.000 description 1
- VSRXQHXAPYXROS-UHFFFAOYSA-N azanide;cyclobutane-1,1-dicarboxylic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OC(=O)C1(C(O)=O)CCC1 VSRXQHXAPYXROS-UHFFFAOYSA-N 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- MMIMIFULGMZVPO-UHFFFAOYSA-N benzyl 3-bromo-2,6-dinitro-5-phenylmethoxybenzoate Chemical compound [O-][N+](=O)C1=C(C(=O)OCC=2C=CC=CC=2)C([N+](=O)[O-])=C(Br)C=C1OCC1=CC=CC=C1 MMIMIFULGMZVPO-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- QLTSDROPCWIKKY-PMCTYKHCSA-N beta-D-glucosaminyl-(1->4)-beta-D-glucosamine Chemical group O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O)[C@@H](CO)O1 QLTSDROPCWIKKY-PMCTYKHCSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229960000997 bicalutamide Drugs 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical group 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 229960003261 carmofur Drugs 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 235000013477 citrulline Nutrition 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000000562 conjugate Substances 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960003901 dacarbazine Drugs 0.000 description 1
- 229960000640 dactinomycin Drugs 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- NDLOVDOICXITOK-UHFFFAOYSA-N dbco-acid Chemical compound OC(=O)CCC(=O)N1CC2=CC=CC=C2C#CC2=CC=CC=C12 NDLOVDOICXITOK-UHFFFAOYSA-N 0.000 description 1
- 230000003413 degradative effect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- XCDIRYDKECHIPE-QHEQPUDQSA-N docetaxel trihydrate Chemical compound O.O.O.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 XCDIRYDKECHIPE-QHEQPUDQSA-N 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- ZWAOHEXOSAUJHY-ZIYNGMLESA-N doxifluridine Chemical compound O[C@@H]1[C@H](O)[C@@H](C)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ZWAOHEXOSAUJHY-ZIYNGMLESA-N 0.000 description 1
- 229950005454 doxifluridine Drugs 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 229950011487 enocitabine Drugs 0.000 description 1
- 229950002973 epitiostanol Drugs 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229960001842 estramustine Drugs 0.000 description 1
- FRPJXPJMRWBBIH-RBRWEJTLSA-N estramustine Chemical compound ClCCN(CCCl)C(=O)OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 FRPJXPJMRWBBIH-RBRWEJTLSA-N 0.000 description 1
- 229960005420 etoposide Drugs 0.000 description 1
- 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 1
- 229960005304 fludarabine phosphate Drugs 0.000 description 1
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 1
- MKXKFYHWDHIYRV-UHFFFAOYSA-N flutamide Chemical compound CC(C)C(=O)NC1=CC=C([N+]([O-])=O)C(C(F)(F)F)=C1 MKXKFYHWDHIYRV-UHFFFAOYSA-N 0.000 description 1
- 229960002074 flutamide Drugs 0.000 description 1
- VVIAGPKUTFNRDU-ABLWVSNPSA-N folinic acid Chemical compound C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-ABLWVSNPSA-N 0.000 description 1
- 235000008191 folinic acid Nutrition 0.000 description 1
- 239000011672 folinic acid Substances 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 1
- 229960005144 gemcitabine hydrochloride Drugs 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 150000002334 glycols Chemical class 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 229960003690 goserelin acetate Drugs 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 229940022353 herceptin Drugs 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000001794 hormone therapy Methods 0.000 description 1
- 235000001050 hortel pimenta Nutrition 0.000 description 1
- DKAGJZJALZXOOV-UHFFFAOYSA-N hydrate;hydrochloride Chemical compound O.Cl DKAGJZJALZXOOV-UHFFFAOYSA-N 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 229960001330 hydroxycarbamide Drugs 0.000 description 1
- 229960001101 ifosfamide Drugs 0.000 description 1
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 229960000779 irinotecan hydrochloride Drugs 0.000 description 1
- GURKHSYORGJETM-WAQYZQTGSA-N irinotecan hydrochloride (anhydrous) Chemical compound Cl.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 GURKHSYORGJETM-WAQYZQTGSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229960001691 leucovorin Drugs 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229960002985 medroxyprogesterone acetate Drugs 0.000 description 1
- PSGAAPLEWMOORI-PEINSRQWSA-N medroxyprogesterone acetate Chemical compound C([C@@]12C)CC(=O)C=C1[C@@H](C)C[C@@H]1[C@@H]2CC[C@]2(C)[C@@](OC(C)=O)(C(C)=O)CC[C@H]21 PSGAAPLEWMOORI-PEINSRQWSA-N 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 description 1
- 229950009246 mepitiostane Drugs 0.000 description 1
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 description 1
- 229960001428 mercaptopurine Drugs 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Chemical group 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 229960000350 mitotane Drugs 0.000 description 1
- ZAHQPTJLOCWVPG-UHFFFAOYSA-N mitoxantrone dihydrochloride Chemical compound Cl.Cl.O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO ZAHQPTJLOCWVPG-UHFFFAOYSA-N 0.000 description 1
- 229960004169 mitoxantrone hydrochloride Drugs 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- BLCLNMBMMGCOAS-UHFFFAOYSA-N n-[1-[[1-[[1-[[1-[[1-[[1-[[1-[2-[(carbamoylamino)carbamoyl]pyrrolidin-1-yl]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-[(2-methylpropan-2-yl)oxy]-1-oxopropan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amin Chemical compound C1CCC(C(=O)NNC(N)=O)N1C(=O)C(CCCN=C(N)N)NC(=O)C(CC(C)C)NC(=O)C(COC(C)(C)C)NC(=O)C(NC(=O)C(CO)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C(CC=1NC=NC=1)NC(=O)C1NC(=O)CC1)CC1=CC=C(O)C=C1 BLCLNMBMMGCOAS-UHFFFAOYSA-N 0.000 description 1
- 229950007221 nedaplatin Drugs 0.000 description 1
- QZGIWPZCWHMVQL-UIYAJPBUSA-N neocarzinostatin chromophore Chemical compound O1[C@H](C)[C@H](O)[C@H](O)[C@@H](NC)[C@H]1O[C@@H]1C/2=C/C#C[C@H]3O[C@@]3([C@@H]3OC(=O)OC3)C#CC\2=C[C@H]1OC(=O)C1=C(O)C=CC2=C(C)C=C(OC)C=C12 QZGIWPZCWHMVQL-UIYAJPBUSA-N 0.000 description 1
- 229960001420 nimustine Drugs 0.000 description 1
- VFEDRRNHLBGPNN-UHFFFAOYSA-N nimustine Chemical compound CC1=NC=C(CNC(=O)N(CCCl)N=O)C(N)=N1 VFEDRRNHLBGPNN-UHFFFAOYSA-N 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 150000002918 oxazolines Chemical class 0.000 description 1
- 238000007248 oxidative elimination reaction Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 229960002340 pentostatin Drugs 0.000 description 1
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 1
- QIMGFXOHTOXMQP-GFAGFCTOSA-N peplomycin Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCCN[C@@H](C)C=1C=CC=CC=1)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1NC=NC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C QIMGFXOHTOXMQP-GFAGFCTOSA-N 0.000 description 1
- 229950003180 peplomycin Drugs 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 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
- 239000007981 phosphate-citrate buffer Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 108010001062 polysaccharide-K Proteins 0.000 description 1
- 229960004293 porfimer sodium Drugs 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 229960001586 procarbazine hydrochloride Drugs 0.000 description 1
- 230000009145 protein modification Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 239000001990 protein-drug conjugate Substances 0.000 description 1
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 229960002185 ranimustine Drugs 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 229950010372 sobuzoxane Drugs 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 108090000250 sortase A Proteins 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- FQZYTYWMLGAPFJ-OQKDUQJOSA-N tamoxifen citrate Chemical compound [H+].[H+].[H+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 FQZYTYWMLGAPFJ-OQKDUQJOSA-N 0.000 description 1
- 229960003454 tamoxifen citrate Drugs 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- 229960002190 topotecan hydrochloride Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 125000000430 tryptophan group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C2=C([H])C([H])=C([H])C([H])=C12 0.000 description 1
- 201000007423 tubular adenocarcinoma Diseases 0.000 description 1
- 229950009811 ubenimex Drugs 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- KDQAABAKXDWYSZ-PNYVAJAMSA-N vinblastine sulfate Chemical compound OS(O)(=O)=O.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 KDQAABAKXDWYSZ-PNYVAJAMSA-N 0.000 description 1
- 229960004982 vinblastine sulfate Drugs 0.000 description 1
- AQTQHPDCURKLKT-JKDPCDLQSA-N vincristine sulfate Chemical compound OS(O)(=O)=O.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=O)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 AQTQHPDCURKLKT-JKDPCDLQSA-N 0.000 description 1
- 229960002110 vincristine sulfate Drugs 0.000 description 1
- 229960005212 vindesine sulfate Drugs 0.000 description 1
- 229960002166 vinorelbine tartrate Drugs 0.000 description 1
- GBABOYUKABKIAF-IWWDSPBFSA-N vinorelbinetartrate Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC(C23[C@H]([C@@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-IWWDSPBFSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 229950009268 zinostatin Drugs 0.000 description 1
Images
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
本発明は、乳がんまたは胃がんの治療のための抗体-薬物複合体に関する。 The present invention relates to antibody-drug conjugates for the treatment of breast cancer or gastric cancer.
均一な抗体-薬物複合体の作製を行うために、抗体と薬物を結合させるリンカーの開発
など様々な手法が試みられている(非特許文献１等を参照)。多くは、リジンアミノ基への
カルボン酸との縮合によるアミド結合形成、あるいは、システインのスルフヒドリル基と
マレイミドのマイケル付加反応、ジスルフィド結合形成反応を介しての結合形成により、
薬物が抗体側に結合される。抗体は、複数のリジン、システインを含むので、薬物結合の
位置と数を制御することができず、不均一になる。また、可変部位のリジンやシステイン
に薬物が結合すると、抗原への結合能力が低下する。
In order to prepare a uniform antibody-drug conjugate, various techniques have been attempted, such as development of a linker that binds an antibody and a drug (see Non-Patent
A drug is attached to the antibody side. Since antibodies contain multiple lysines and cysteines, the position and number of drug attachments cannot be controlled, resulting in heterogeneity. In addition, when a drug binds to lysine or cysteine in the variable region, the ability to bind to an antigen is reduced.
アミノ基は、抗体に約９０含まれており、そのうちの３０は、容易に修飾されることが
知られている。ヒトIgG1の場合、鎖間ジスルフィド結合は４つあり、その結合を担うシス
テインは８個存在する。従って、アミノ基やチオール基を介した修飾を行うと、多くの異
性体を生じることになる。特に、正電荷を有するリジン側鎖は通常抗体の分子表面に存在
するために、疎水性の高い薬物をリジン残基を介して結合させると、薬物動態や体内クリ
アランスに影響を与え得るため、リジン残基を利用した薬物導入は回避されることが多い
。
Antibodies contain approximately 90 amino groups, 30 of which are known to be readily modified. In human IgG1, there are four interchain disulfide bonds and eight cysteines responsible for these bonds. Therefore, modification via an amino group or a thiol group will give rise to many isomers. In particular, positively charged lysine side chains are usually present on the molecular surface of antibodies, so binding highly hydrophobic drugs via lysine residues can affect pharmacokinetics and body clearance. Drug introduction via residues is often avoided.
また、抗体の一部のアミノ酸残基をアジド基のような官能基をもつ非天然型アミノ酸に
置換する方法もあるが、抗体をin vitro protein synthesis により発現させる労力が必
要である(非特許文献２等を参照)。
There is also a method of substituting some of the amino acid residues of the antibody with a non-natural amino acid having a functional group such as an azide group. 2, etc.).
また、ある特定のペプチド配列を用いて、sortase A, transglutaminase, formylglyci
ne converting enzyme を用いて部位特異的に薬物を結合することもできる。それぞれ、L
eu-Pro-X-Thr-Gly (X = 任意のアミノ酸)、Leu-Leu-Gln-Gly-Ala を認識する。しかしタ
ンパク質に該当する配列が存在しない場合には、付加できない（非特許文献３等を参照）
。
Also, using a certain peptide sequence, sortase A, transglutaminase, formylglyci
It is also possible to bind drugs site-specifically using ne converting enzymes. respectively, L.
Recognizes eu-Pro-X-Thr-Gly (X = any amino acid), Leu-Leu-Gln-Gly-Ala. However, if the sequence corresponding to the protein does not exist, it cannot be added (see Non-Patent
.
一方で、抗体はAsn297位に普遍的にN-結合型糖鎖を持つ。N-結合型糖鎖はその複雑な生
合成過程から微細構造が少しずつ異なる。抗体医薬において、糖鎖構造が活性に影響を及
ぼすことが知られているが(非特許文献４等)、抗体作製時に糖鎖構造を均一にすることは
困難である。
On the other hand, antibodies universally have an N-linked sugar chain at Asn297. N-linked glycans have slightly different fine structures due to their complicated biosynthetic processes. In antibody drugs, it is known that the sugar chain structure affects the activity (Non-Patent
特異的に反応する部位を持つ修飾糖ユニットを糖供与体として、改変したガラクトース
転移酵素、シアル酸転移酵素を用いて、糖ユニットをひとつずつ延長する糖転移酵素を用
いて、抗体の糖鎖部に薬物を結合させる手法も用いられるが、糖鎖構造が不均一であるた
めに、最終的な抗体-薬物複合体が不均一になったり、複数回の糖鎖伸長反応が必要であ
ったりする問題点がある（非特許文献５等を参照）。また、非ヒト型糖鎖を排除すること
はできない。また、薬物と特異的に反応可能である修飾基を持つ糖供与体である糖ヌクレ
オチドの合成は数工程必要であり、全収率は高くなく、中間体などは不安定である。
Using a modified sugar unit with a specific reaction site as a sugar donor, modified galactosyltransferase and sialyltransferase are used to extend the sugar units one by one, using a glycosyltransferase to extend the sugar chain portion of the antibody. Although a method of conjugating a drug to the antibody is also used, the heterogeneity of the sugar chain structure results in a heterogeneous final antibody-drug conjugate and the need for multiple sugar chain elongation reactions. There is a problem (see Non-Patent Document 5, etc.). Also, non-human sugar chains cannot be excluded. In addition, the synthesis of sugar nucleotides, which are sugar donors having a modifying group that can specifically react with drugs, requires several steps, the overall yield is not high, and intermediates and the like are unstable.
また、糖鎖の水酸基をNaIO4により酸化開裂して、アルデヒド基を創出し、ヒドラジド
と反応させて、位置特異的に反応させることも行われている。しかしながら、糖鎖構造が
不均一であることから、結合の数を制御することはできない。また、トリプトファン、シ
ステイン、メチオニンなど酸化に不安定な官能基を含むアミノ酸残基に対する影響は無視
できない（非特許文献６等を参照）。
In addition, oxidative cleavage of hydroxyl groups of sugar chains with NaIO 4 creates aldehyde groups, which are then reacted with hydrazides for regiospecific reactions. However, the number of bonds cannot be controlled due to the heterogeneity of the sugar chain structure. In addition, the effect on amino acid residues containing oxidation-labile functional groups such as tryptophan, cysteine, and methionine cannot be ignored (see Non-Patent Document 6, etc.).
細胞内の代謝を利用して、官能基修飾したフコースなどを導入し、抗体糖鎖に修飾フコ
ースを導入することが可能である。フコースの官能基を利用して、部位特異的に薬物を結
合することが可能である（非特許文献７等を参照）。
Utilizing intracellular metabolism, functional group-modified fucose or the like can be introduced, and modified fucose can be introduced into antibody sugar chains. It is possible to site-specifically bind a drug using the functional group of fucose (see Non-Patent
上記の問題点を解決するために、抗体にもともと存在する糖鎖をAsnに結合しているN-
アセチルグルコサミン１残基のみを残して切断し（糖鎖微細構造、非ヒト型糖鎖は消失す
る）、一挙に特異的反応を起こすような官能基（例えば、アジド基）を持つ、均一な構造
のヒト型糖鎖を改変糖加水分解酵素を用いて転移させる（糖鎖部分が均一となる）。特異
的反応をおこす官能基（例えば、アジド基）と反応する官能基を結合した薬物を付加する
と、均一構造を持つタンパク質-薬物複合体を作製できる。糖鎖は普遍的なタンパク質修
飾であるので、sortaseやtransglutaminaseのような酵素が要求する特定のアミノ酸配列
を必要とすることなく、コンジュゲートが可能である。改変糖加水分解酵素を用いてアジ
ド基などの官能基をもつ糖鎖を抗体に導入した例も存在する（非特許文献８等を参照）。
In order to solve the above problems, N-
Uniform structure with a functional group (e.g., azide group) that causes a specific reaction at once by cutting leaving only one residue of acetylglucosamine (the sugar chain fine structure and non-human sugar chain disappear) The human-type sugar chain of is transferred using a modified glycohydrolase (the sugar chain portion becomes uniform). A protein-drug conjugate with a uniform structure can be produced by adding a drug that binds a functional group that reacts with a functional group that causes a specific reaction (eg, an azide group). Since sugar chains are universal protein modifications, conjugation is possible without the need for specific amino acid sequences required by enzymes such as sortase and transglutaminase. There are also examples in which a sugar chain having a functional group such as an azide group is introduced into an antibody using a modified glycohydrolase (see Non-Patent Document 8, etc.).
薬物とリンカーを結合する反応は、アジドとひずんだアルキンとのクリック反応や逆電
子要請型Diels-Alder反応の用な生物直交的反応が使用できる。
Bioorthogonal reactions such as the click reaction between azide and strained alkyne and the inverse electron demand Diels-Alder reaction can be used to link the drug to the linker.
リンカーとして、カテプシン切断型リンカーについても報告されている（非特許文献９
等を参照）。
As a linker, a cathepsin-cleavable linker has also been reported (Non-Patent Document 9
etc.).
本発明は、抗HER2抗体と抗がん剤の抗体-薬物複合体を含む乳がんまたは胃がん治療剤
の提供を目的とする。
An object of the present invention is to provide a therapeutic agent for breast cancer or gastric cancer comprising an antibody-drug conjugate of an anti-HER2 antibody and an anticancer agent.
本発明者らは、HER2を発現しているがんの治療剤として、抗HER2抗体と抗がん剤である
薬物を含む抗体-薬物複合体（ADC)の治療効果を向上させる方法について鋭意検討を行っ
た。in vitro でのHER2を発現するがん細胞に対する効果の検討において、抗体に薬物を
糖鎖とカテプシン切断型リンカーを介して結合させたADCが薬物単独と同等の殺細胞効果
を奏するという予測できない結果が得られた。また、糖鎖を介して薬物を付加することに
より、均一なADCが得られることを見出した。これらの結果より、抗HER2抗体に薬物を糖
鎖およびカテプシン切断型リンカーを介して付加したADCが乳がんまたは胃がん、特に乳
がん治療に有効であることを見出し本発明を完成させるに至った。
The present inventors diligently studied methods for improving the therapeutic effects of antibody-drug conjugates (ADCs) containing an anti-HER2 antibody and an anticancer drug as therapeutic agents for HER2-expressing cancers. did In vitro studies of the effect on HER2-expressing cancer cells showed an unexpected result that an ADC, in which a drug was conjugated to an antibody via a sugar chain and a cathepsin-cleavable linker, exhibited a cell-killing effect equivalent to that of the drug alone. was gotten. We also found that a uniform ADC can be obtained by adding a drug via a sugar chain. Based on these results, the inventors have found that an ADC obtained by attaching a drug to an anti-HER2 antibody via a sugar chain and a cathepsin-cleavable linker is effective in treating breast cancer or gastric cancer, particularly breast cancer, and completed the present invention.
すなわち、本発明は以下のとおりである。
[１] 抗HER2抗体に抗がん剤がカテプシン切断型リンカーであるバリン-シトルリン(Val-
Cit)ペプチドリンカーおよび糖鎖を介して付加されており、細胞中でカテプシンによりリ
ンカーが切断され、抗がん剤が放出される、抗体-薬物複合体。
[２] 抗がん剤がアウリスタチンである、[１]の抗体-薬物複合体。
[３] アウリスタチンが、monomethyl auristatin E (MMAE)、monomethyl auristatin D
(MMAD)およびmonomethyl auristatin F (MMAF)からなる群から選択される、[２]の抗体-
薬物複合体。
[４] 抗HER2抗体が均一糖鎖構造を有する糖鎖改変した抗体である、[１]～[３]のいずれ
かの抗体-薬物複合体。
[５] 抗体がトラスツズマブである、[１]～[４]のいずれかの抗体-薬物複合体。
[６] [１]～[５]のいずれかの抗体-薬物複合体を含む、乳がんまたは胃がん治療剤。
[７] 他の抗がん剤と併用される、[６]の乳がんまたは胃がん治療剤。
That is, the present invention is as follows.
[1] Valine-citrulline (Val-
Cit) an antibody-drug conjugate attached via a peptide linker and a sugar chain, where the linker is cleaved by cathepsins in the cell, releasing the anticancer drug.
[2] The antibody-drug conjugate of [1], wherein the anticancer agent is auristatin.
[3] Auristatin is monomethyl auristatin E (MMAE), monomethyl auristatin D
the antibody of [2] selected from the group consisting of (MMAD) and monomethyl auristatin F (MMAF)-
drug complex.
[4] The antibody-drug conjugate of any one of [1] to [3], wherein the anti-HER2 antibody is a sugar chain-engineered antibody having a uniform sugar chain structure.
[5] The antibody-drug conjugate of any one of [1] to [4], wherein the antibody is trastuzumab.
[6] A therapeutic agent for breast cancer or gastric cancer, comprising the antibody-drug conjugate of any one of [1] to [5].
[7] The therapeutic agent for breast cancer or gastric cancer of [6], which is used in combination with other anticancer agents.
抗HER2抗体に薬物を糖鎖およびカテプシン切断型リンカーを介して付加したADCは、HER
2を発現するがんの中でも、乳がんまたは胃がんに対して顕著な抗がん効果を示す。
ADCs, in which drugs are attached to anti-HER2 antibodies via sugar chains and cathepsin-cleavable linkers,
Among cancers that express 2, it exhibits remarkable anticancer effects against breast cancer and gastric cancer.
以下、本発明を詳細に説明する。 The present invention will be described in detail below.
本発明は、がん治療のための抗体-薬物複合体（ADC: Antibody-drug conjugate)である
。本発明のADCにおいて、抗体は抗HER2抗体であり、薬物は抗がん剤である。抗がん剤は
、チューブリン合成阻害剤（抗がん性植物アルカロイド）、トポイソメラーゼ阻害剤、核
酸合成の阻害剤であるアルキル化剤、代謝拮抗剤、抗生物質抗癌剤、ホルモン製剤、白金
製剤、非特異的抗悪性腫瘍剤等の公知の抗がん剤の他新規に開発される抗がん剤を含む。
具体的には、例えば、アウリスタチン、硫酸ビンクリスチン、硫酸ビンブラスチン、硫酸
ビンデシン、ドセタキセル水和物、パクリタキセル、酒石酸ビノレルビン、メイタンシノ
イド等のチューブリン合成阻害剤（抗がん性植物アルカロイド）；カンプトテシン、トポ
テカン、エトポシド、塩酸イリノテカン、塩酸ノギテカン等のトポイソメラーゼ阻害薬；
シクロフォスファミド、イフォスファミド、メルファラン、チオテバ、ブスルファン、カ
ルボコン、ダカルバジン、塩酸ニムスチン、ラニムスチン等のアルキル化剤；メトトレキ
サート、メルカプトプリン、6-メルカプトプリンリボシド、フルオロウラシル(5-FU)、テ
ガフール、テガフールウラシル、カルモフール、ドキシフルリジン、シタラビンオクホス
ファート、ヒドロキシカルバミド、シタラビン、塩酸ゲムシタビン、リン酸フルダラビン
、エノシタビン、ロイコボリン等の代謝拮抗剤；塩酸ドキソルビシン、塩酸イダルビシン
、塩酸エピルビシン、塩酸ピラルビシン、塩酸ダウノルビシン、塩酸アクラルビシン、マ
イトマイシンC、アクチノマイシンD、ブレオマイシン、硫酸ペプロマイシン、ネオカルチ
ノスタチン、ジノスタチンスチマラマー等の抗がん性抗生物質；リン酸エストラムスチン
タトリウム、フルタミド、ビカルタミド、酢酸ゴセレリン、酢酸ニュープロレリン、クエ
ン酸タモキシフェン、塩酸フォドロゾール水和物、アナストロゾール、メピチオスタン、
エピチオスタノール、酢酸メドロキシプロゲステロン等のホルモン製剤；シスプラチン、
カルボプラチン、ネダプラチン等の白金製剤；クレスチン、レンアチン、シゾフィラン、
ウベニメクス等の非特異的抗悪性腫瘍剤；塩酸ミトキサントロン、塩酸プロカルバジン、
ペントスタチン、ソブゾキサン、トレチノイン、L-アスパラギナーゼ、アセグラトン、ミ
トタン、ポルフィマーナトリウム等が挙げられる。
The present invention is an antibody-drug conjugate (ADC) for cancer therapy. In the ADC of the present invention, the antibody is an anti-HER2 antibody and the drug is an anticancer agent. Anticancer agents include tubulin synthesis inhibitors (anticancer plant alkaloids), topoisomerase inhibitors, alkylating agents that are inhibitors of nucleic acid synthesis, antimetabolites, antibiotic anticancer agents, hormone preparations, platinum preparations, non- In addition to known anticancer agents such as specific antineoplastic agents, newly developed anticancer agents are included.
Specifically, for example, tubulin synthesis inhibitors (anticancer plant alkaloids) such as auristatin, vincristine sulfate, vinblastine sulfate, vindesine sulfate, docetaxel hydrate, paclitaxel, vinorelbine tartrate, maytansinoids; Topoisomerase inhibitors such as topotecan, etoposide, irinotecan hydrochloride, topotecan hydrochloride;
Alkylating agents such as cyclophosphamide, ifosfamide, melphalan, thioteva, busulfan, carbocone, dacarbazine, nimustine hydrochloride, ranimustine; methotrexate, mercaptopurine, 6-mercaptopurine riboside, fluorouracil (5-FU), tegafur, tegafur Antimetabolites such as uracil, carmofur, doxifluridine, cytarabine ocphosphate, hydroxycarbamide, cytarabine, gemcitabine hydrochloride, fludarabine phosphate, enocitabine, leucovorin; Anticancer antibiotics such as mitomycin C, actinomycin D, bleomycin, peplomycin sulfate, neocarzinostatin, dinostatin stimaramer; Estramustine tatrium phosphate, flutamide, bicalutamide, goserelin acetate, neuprorelin acetate, tamoxifen citrate, fodrozole hydrochloride hydrate, anastrozole, mepitiostane,
Hormone preparations such as epithiostanol, medroxyprogesterone acetate; cisplatin,
Platinum agents such as carboplatin and nedaplatin; krestin, lenatin, schizophyllan,
non-specific antineoplastic agents such as ubenimex; mitoxantrone hydrochloride, procarbazine hydrochloride,
Pentostatin, sobuzoxane, tretinoin, L-asparaginase, acegratone, mitotane, porfimer sodium and the like.
この中でも、チューブリン合成阻害剤であり、非常に強い抗腫瘍剤であるauristatin（
アウリスタチン）が挙げられる。アウリスタチンには、monomethyl auristatin E (MMAE
、モノメチルアウリスタチンE)、monomethyl auristatin D (MMAD、モノメチルアウリス
タチンD)、monomethyl auristatin F (MMAF、モノメチルアウリスタチンF)等が含まれる
。この中でも、monomethyl auristatin E (MMAE、モノメチルアウリスタチンE)が好まし
い。
Among them, auristatin (auristatin), which is a tubulin synthesis inhibitor and a very strong anti-tumor agent
auristatin). Auristatins include monomethyl auristatin E (MMAE
, monomethyl auristatin E), monomethyl auristatin D (MMAD, monomethyl auristatin D), monomethyl auristatin F (MMAF, monomethyl auristatin F), and the like. Among these, monomethyl auristatin E (MMAE, monomethyl auristatin E) is preferred.
本発明のADCは、乳がんおよび胃がんの治療に用いられる。
HER2は、細胞表面に存在する、EGFR(epidermal growth factor receptor)ファミリーに
属する約185kDaの受容体型チロシンキナーゼであり、上皮成長因子受容体（EGFR)に類似
した構造を有している。HER2は、乳がんや胃がんにおいて過剰に発現することがあること
が知られている。本発明のADCに用いられる抗HER2抗体は、リコンビナント抗HER2モノク
ローナル抗体であり、ヒトの定常領域を有するキメラ抗体、ヒト抗体のCDR（相補性決定
領域）をヒト以外の動物の抗HER2抗体のCDRで移植したヒト化抗体（CDR移植抗体）、ヒト
由来の抗体遺伝子の発現産物であるヒト抗体が含まれるが、ヒト化抗体やヒト抗体が好ま
しい。完全ヒトモノクローナル抗体は、ヒト免疫グロブリン重鎖および軽鎖遺伝子座の大
部分が遺伝子導入されたマウスを免疫することにより作製することができる。抗体のクラ
スは好ましくはIgG1である。好ましい抗HER2抗体として、ヒト化IgG1抗体であるトラスツ
ズマブ（Trastuzumab）（ハーセプチン（登録商標））が挙げられる。
The ADCs of the invention are used in the treatment of breast cancer and gastric cancer.
HER2 is a receptor-type tyrosine kinase of about 185 kDa belonging to the EGFR (epidermal growth factor receptor) family present on the cell surface, and has a structure similar to that of epidermal growth factor receptor (EGFR). HER2 is known to be overexpressed in breast cancer and gastric cancer. The anti-HER2 antibody used in the ADC of the present invention is a recombinant anti-HER2 monoclonal antibody, a chimeric antibody having a human constant region, a human antibody CDR (complementarity determining region), and a non-human animal anti-HER2 antibody CDR. Humanized antibodies (CDR-grafted antibodies) grafted with , and human antibodies that are expression products of human-derived antibody genes are included, but humanized antibodies and human antibodies are preferred. Fully human monoclonal antibodies can be produced by immunizing mice transgenic for large portions of human immunoglobulin heavy and light chain loci. The antibody class is preferably IgG1. Preferred anti-HER2 antibodies include Trastuzumab (Herceptin®), a humanized IgG1 antibody.
本発明のADCにおいては、薬物は抗体のアミノ基やシステイン基を介して抗体に付加し
ているのではなくて、抗体の定常領域に結合している糖鎖を介して付加している。
In the ADC of the present invention, the drug is not attached to the antibody via the amino group or cysteine group of the antibody, but is attached via the sugar chain bound to the constant region of the antibody.
構造が均一なADCの作製は、活性薬物活性、薬物動態を制御するために必須であり、レ
ギュラトリー・サイエンスからもその重要性が指摘され、ADC開発の中心的議題である。A
DCは、薬物（イメージング剤、RI、siRNA等も含む）を抗体に結合させ、体内でがん等の
病変部位に選択的に送達させ、薬物を病変部位のみにて放出する（イメージング剤の場合
には放出させない）ことにより、効果の有効発現と副作用の低減を狙うDDS（Drug delive
ry system）技術のひとつである。
The production of ADCs with uniform structure is essential for controlling active drug activity and pharmacokinetics, and its importance has been pointed out by regulatory science, and is a central issue in ADC development. A.
DCs bind drugs (including imaging agents, RI, siRNA, etc.) to antibodies, selectively deliver them to lesions such as cancer in the body, and release the drugs only at lesions (in the case of imaging agents DDS (drug deliver
ry system) technology.
一方、抗体に代表されるタンパク質には、不均一な構造の糖鎖が付加しており、糖鎖構
造により生理活性も異なることが知られている。例えば、ポテリジオ(登録商標）（フコ
ース除去型抗CCR4抗体）にみられるように、糖鎖構造を改変することにより、ADCC活性が
100倍に上昇した例も知られている。また、抗体作製にあたりCHO細胞等を使用すると、抗
体に非ヒト型糖鎖が付加され、その抗原性からアナフィラキシーショックを起こすことが
ある。
On the other hand, proteins typified by antibodies are known to have heterogeneously-structured sugar chains attached thereto, and that their physiological activities vary depending on the sugar chain structure. For example, as seen in POTELIGEO (registered trademark) (fucose-removed anti-CCR4 antibody), ADCC activity can be enhanced by modifying the sugar chain structure.
A 100-fold increase is also known. In addition, when CHO cells or the like are used for antibody production, non-human sugar chains are added to the antibody, which may cause anaphylactic shock due to its antigenicity.
これら２つの問題点を解決するために、抗体が本来持っている糖鎖を切断、再構成する
ことにより、糖鎖構造を制御しつつ、糖鎖を介して、構造均一であるヒト型糖鎖を持つ抗
体-薬物複合体を作製することができる。この方法により、in vitroにおけるタンパク質
合成などの分子生物学的手法を使用することなく、薬物を抗体に位置特異的に結合させ、
薬物結合数を制御することが可能になる。
In order to solve these two problems, by cleaving and reconstituting the sugar chain originally possessed by the antibody, a human-type sugar chain with a uniform structure is obtained through the sugar chain while controlling the sugar chain structure. can be made antibody-drug conjugates with By this method, the drug can be site-specifically attached to the antibody without using molecular biological techniques such as in vitro protein synthesis,
It becomes possible to control the number of drug bindings.
本発明のADCにおいては、糖鎖構造が均一となるように、薬物を糖鎖を介して抗体に付
加してある。
In the ADC of the present invention, drugs are attached to antibodies via sugar chains so that the sugar chain structure is uniform.
本発明のADCの作製方法を以下に例示する。
（１） 元々糖鎖構造が不均一な抗HER2抗体の糖鎖を切断する。糖鎖の切断は、エンド-
β-N-アセチルグルコサミニダーゼ（EndoS）を用いて行うことができる。EndoSにより、
抗体のFc領域のCHドメインに位置する297番目のアスパラギン（Asn)に結合しているN型複
合型糖鎖の還元末端のN-アセチルグルコサミン（GlcNAc)１残基を残して切断される。還
元末端のN-アセチルグルコサミンには、フコースが結合していても、結合していなくても
よい。この後、クロマトグラフィーを用いて、糖鎖が切断された抗体を精製すればよい。
クロマトグラフィーとしては、ゲル濾過クロマトグラフィー、アフィニティークロマトグ
ラフィー、イオン交換クロマトグラフィー、疎水性クロマトグラフィー等が挙げられる。
A method for producing the ADC of the present invention is exemplified below.
(1) Cleave the sugar chains of the anti-HER2 antibody, which originally has a heterogeneous sugar chain structure. Cleavage of sugar chains is endo-
It can be performed using β-N-acetylglucosaminidase (EndoS). By EndoS,
The N-type complex-type sugar chain bound to the 297th asparagine (Asn) located in the CH domain of the Fc region of the antibody is cleaved leaving one N-acetylglucosamine (GlcNAc) residue at the reducing end. N-acetylglucosamine at the reducing end may or may not be bound with fucose. After that, chromatography may be used to purify the antibody in which the sugar chain has been cleaved.
Chromatography includes gel filtration chromatography, affinity chromatography, ion exchange chromatography, hydrophobic chromatography and the like.
（２） 次いで、糖鎖を切断した抗体に、還元末端にオキサゾリン化されたGlcNAcを有し
、反対側の末端にアジド基（N3)を付加した均一なオキサゾリン糖鎖(アジドオキサゾリン
糖鎖）をドナーとして、アクセプターである（１）で得られた糖鎖を切断した抗体のN-ア
セチルグルコサミン（GlcNAc)残基に糖鎖を結合させる。この反応においては、抗体のN-
アセチルグルコサミン（GlcNAc)残基の４位にドナーのオキサゾリン環が反応し、キトビ
オース構造を形成し結合する。この際、例えば、糖鎖切断活性を抑制し、糖鎖転移活性を
向上させたEndoSのD233Q変異体を用いればよい。この工程により抗体へ均一な糖鎖を付加
させることができる。この後、クロマトグラフィーにより糖鎖が結合させた抗体を精製す
ればよい。このようにして糖鎖を結合させた構造は、均一糖鎖構造を有する糖鎖改変した
抗体である。
(2) Next, a uniform oxazoline sugar chain (azidooxazoline sugar chain) having an oxazolated GlcNAc at the reducing end and an azide group (N 3 ) added to the opposite end of the antibody with the sugar chain cleaved is used as a donor to bind a sugar chain to the acceptor N-acetylglucosamine (GlcNAc) residue of the sugar chain-cleaved antibody obtained in (1). In this reaction, the N-
The donor oxazoline ring reacts with the 4-position of the acetylglucosamine (GlcNAc) residue to form a chitobiose structure and bind. In this case, for example, an EndoS D233Q mutant with suppressed glycosylation activity and improved glycosylation activity may be used. This step allows uniform sugar chains to be added to the antibody. After that, the antibody bound to the sugar chain may be purified by chromatography. The structure to which sugar chains are bound in this way is a sugar chain-engineered antibody having a uniform sugar chain structure.
抗体に結合させる均一な構造の糖鎖は目的に応じて適宜選択されるが、還元末端のN-ア
セチルグルコサミン（GlcNAc)を含め、ジアセチルキトビオース(GlcNAc-GlcNAc)にマンノ
ース（Man)のオリゴマーが結合した構造を有する高マンノース型糖鎖、ジアセチルキトビ
オースにMan並びにGlcNAc、ガラクトース（Gal)、シアル酸（Neu5Ac)の少なくとも１つが
結合した複合（コンプレックス）型糖鎖、ジアセチルキトビオースに高マンノース型と複
合型が混成した糖鎖構造を有する混成（ハイブリッド）型糖鎖が挙げられ、シアル酸の有
無、コアフコースの有無、分岐鎖の有無等により種々の構造の糖鎖がある。
A sugar chain with a uniform structure to be bound to an antibody is appropriately selected depending on the purpose. A high mannose type sugar chain having a structure in which is bound to diacetylchitobiose, a complex type sugar chain in which at least one of Man and GlcNAc, galactose (Gal), sialic acid (Neu5Ac) is bound to diacetylchitobiose, to diacetylchitobiose Examples include hybrid type sugar chains having a sugar chain structure in which a high-mannose type and a complex type are mixed, and there are sugar chains with various structures depending on the presence or absence of sialic acid, the presence or absence of core fucose, the presence or absence of branched chains, and the like.
代表的な糖鎖として、図１に示す構造を有する糖鎖が挙げられる。図１に示す糖鎖の還
元末端のN-アセチルグルコサミン（GlcNAc)には、フコースが結合していてもよい。
A representative sugar chain includes a sugar chain having the structure shown in FIG. Fucose may be bound to N-acetylglucosamine (GlcNAc) at the reducing end of the sugar chain shown in FIG.
（３） 次いで、抗体に結合させたアジド基を有する糖鎖にmonomethyl auristatin E (M
MAE)の薬物をリンカーを介して結合させる。この際、アジド基を有する糖鎖のアジド基に
リンカーと結合させた薬物を結合させればよい。リンカーはカテプシン等のシステインプ
ロテアーゼ酵素により切断されるリンカー、すなわちカテプシン切断型リンカーを用いれ
ばよい。このようなリンカーとして、バリンとシトルリンを結合させたバリン-シトルリ
ン(Val-Cit)ペプチドリンカーが挙げられる。アジド基とリンカーと結合させた薬物との
結合は、リンカーと薬物の結合体のリンカー側にWSCDI等の環状アルキンをクリック反応
により結合させればよい。この後、クロマトグラフィーにより生成物を精製すればよい。
(3) Then, monomethyl auristatin E (M
MAE) drug is bound via a linker. At this time, the drug bound to the linker may be bound to the azide group of the sugar chain having the azide group. A linker that is cleaved by a cysteine protease enzyme such as cathepsin, ie, a cathepsin-cleavable linker may be used as the linker. Such linkers include valine-citrulline (Val-Cit) peptide linkers, which combine valine and citrulline. The binding between the azide group and the drug bound to the linker can be achieved by binding a cyclic alkyne such as WSCDI to the linker side of the linker-drug conjugate by click reaction. After this, the product may be purified by chromatography.
図２に本発明のADCの作製方法の概略を示す。
このようにして得られたADCは、抗HER2抗体に薬物がカテプシン切断型リンカーを介し
て付加されている。
FIG. 2 shows an outline of the method for producing the ADC of the present invention.
The ADC thus obtained has an anti-HER2 antibody attached with a drug via a cathepsin-cleavable linker.
本発明のADCは、薬物を糖鎖改変抗体の特定の部位の糖鎖に結合させるため、均一な構
造を有している。また、抗体に結合させる糖鎖の分岐鎖の数に応じて、１分子の抗体に対
して、２～８分子の薬物を結合させることができる。例えば、図1に示す糖鎖を結合させ
た場合、１分子の抗体に対して、４分子の薬物が結合する（DAR:Drug Antibody ratio =
4)。
The ADC of the present invention has a uniform structure in order to bind a drug to a sugar chain at a specific site of a sugar chain-modified antibody. In addition, depending on the number of branched sugar chains to be bound to the antibody, 2 to 8 drug molecules can be bound to one molecule of the antibody. For example, when the sugar chains shown in Fig. 1 are bound, four drug molecules bind to one antibody molecule (DAR: Drug Antibody ratio =
Four).
本発明のADCは、その塩、又はそれらの水和物も包含する。
ADCを被験体に投与した場合、ADCは乳がんおよび胃がん細胞に発現しているHER2に結合
し、ADCとHER2の複合体が細胞内に取り込まれる。がん細胞中に存在するカテプシンによ
りカテプシン切断型リンカーが切断され、細胞内でMMAEが放出される。その後、MMAEの作
用でがん細胞が死滅する。
ADCs of the present invention also include salts thereof or hydrates thereof.
When ADC is administered to a subject, ADC binds to HER2 expressed in breast cancer and gastric cancer cells, and the complex of ADC and HER2 is taken up into cells. Cathepsins present in cancer cells cleave the cathepsin-cleavable linker, releasing MMAE intracellularly. The cancer cells are then killed by the action of MMAE.
本発明のADCは、乳がんまたは胃がんの治療に有用であり、特に乳がん治療に有用であ
る。本発明のADCはカテプシン切断型リンカーを有しており、細胞中でのADCからの薬物の
放出は細胞内のカテプシンの作用により行われる。カテプシンはがん細胞で発現が向上し
ているが、特に乳がんで発現が向上している可能性があり、乳がん細胞中で薬物が効率的
に放出される可能性がある。
The ADCs of the present invention are useful in treating breast cancer or gastric cancer, and are particularly useful in treating breast cancer. The ADC of the present invention has a cathepsin-cleavable linker, and release of the drug from the ADC in cells is achieved by the action of intracellular cathepsins. Cathepsins are upregulated in cancer cells, and may be upregulated, especially in breast cancer, which may lead to efficient drug release in breast cancer cells.
ADCが抗がん効果を発揮するためには、ADCががん細胞に到達し取り込まれる工程とがん
細胞中で薬物を放出する工程があり、ADCの抗がん効果はこの２つの工程の効率性を考慮
する必要がある。従来は、これら２つの工程の効率性を評価することができず、ADCの設
計において、抗体、薬物、リンカーの組合せを適切に決め、ADCを設計し、さらに効果が
高いがん種を特定することはできなかった。本発明においては、HER2の発現が高いがん（
乳がん、胃がん）を対象として、抗体として抗HER抗体を用い、薬物としてMMAEを用いて
、リンカーとして、カテプシン切断型リンカーを用いることにより、乳がんに特に効果の
高いADCを作製することができた。
In order for ADCs to exert their anticancer effects, there are two processes: the process in which ADCs reach cancer cells and are taken up, and the process in which they release drugs in cancer cells. Efficiency must be considered. Conventionally, it is not possible to evaluate the efficiency of these two processes, so in ADC design, we need to appropriately determine the combination of antibody, drug, and linker, design the ADC, and identify the cancer type that is more effective. I couldn't. In the present invention, cancers with high HER2 expression (
Breast cancer, gastric cancer), using an anti-HER antibody as an antibody, using MMAE as a drug, and using a cathepsin-cleavable linker as a linker, we were able to prepare an ADC that is particularly effective against breast cancer.
本発明は、上記ADCを有効成分として含む抗乳がん剤および抗胃がん剤も包含する。
本発明のADCを含む抗乳がん剤および抗胃がん剤の投与形態は限定されず、経口、非経
口等により投与することができる。非経口投与として、静脈内、皮下、筋肉内、腹腔内注
射等が挙げられる。製剤の投与形態としては、経口液剤、錠剤、カプセル剤、丸剤、散剤
等が挙げられる。カプセル剤、錠剤、散剤、顆粒剤などは、乳糖、ブドウ糖、ショ糖、マ
ンニトールなどの賦形剤；デンプン、アルギン酸ナトリウムなどの崩壊剤；ステアリン酸
マグネシウム、タルクなどの滑沢剤；ポリビニルアルコール、ヒドロキシプロピルセルロ
ース、ゼラチンなどの結合剤；脂肪酸エステルなどの界面活性剤；グリセリンなどの可塑
剤などを添加剤として用いて製造できる。
The present invention also includes an anti-breast cancer agent and an anti-gastric cancer agent containing the ADC as an active ingredient.
The dosage form of the anti-breast cancer agent and anti-gastric cancer agent containing the ADC of the present invention is not limited, and they can be administered orally, parenterally, and the like. Parenteral administration includes intravenous, subcutaneous, intramuscular, intraperitoneal injection and the like. Dosage forms of formulations include oral liquids, tablets, capsules, pills, powders and the like. Capsules, tablets, powders, granules, etc. contain excipients such as lactose, glucose, sucrose, mannitol; disintegrants such as starch and sodium alginate; lubricants such as magnesium stearate and talc; It can be produced using binders such as propylcellulose and gelatin; surfactants such as fatty acid esters; plasticizers such as glycerin, etc., as additives.
乳剤及びシロップ剤のような液体調製物は、水、ショ糖、ソルビトール、果糖などの糖
類；ポリエチレングリコール、プロピレングリコールなどのグリコール類；ごま油、オリ
ーブ油、大豆油などの油類；ｐ－ヒドロキシ安息香酸エステル類などの防腐剤；ストロベ
リーフレーバー、ペパーミントなどのフレーバー類などを添加剤として用いて製造できる
。
Liquid preparations such as emulsions and syrups may contain water, sugars such as sucrose, sorbitol, fructose; glycols such as polyethylene glycol, propylene glycol; oils such as sesame oil, olive oil, soybean oil; Preservatives such as esters; flavors such as strawberry flavor and peppermint can be used as additives for production.
注射剤は、水、ショ糖、ソルビトール、キシロース、トレハロース、果糖などの糖類；
マンニトール、キシリトール、ソルビトールなどの糖アルコール；リン酸緩衝液、クエン
酸緩衝液、グルタミン酸緩衝液などの緩衝液；脂肪酸エステルなどの界面活性剤などを添
加剤として用いることができる。
Injections include water, sugars such as sucrose, sorbitol, xylose, trehalose, and fructose;
Sugar alcohols such as mannitol, xylitol and sorbitol; buffers such as phosphate buffers, citrate buffers and glutamic acid buffers; surfactants such as fatty acid esters and the like can be used as additives.
また、抗HER2抗体により乳がんまたは胃がん細胞を標的として、乳がんまたは胃がん細
胞に効率的に到達するので、全身投与してもよいが、がん組織に局所的に投与してもよい
。
In addition, since the anti-HER2 antibody targets breast cancer or gastric cancer cells and efficiently reaches breast cancer or gastric cancer cells, it may be administered systemically or locally to the cancer tissue.
治療に用いるのに必要な本発明のADCの投与量は、投与する患者の年齢、性別、重篤度
等により変えることができるが、最終的には担当医師が決めることができる。例えば、AD
Cを１回あたり0.05～10mg/kg体重、好ましくは0.1～２mg/kg体重で投与すればよい。また
、MMAE等の薬物当量で、１回当たり0.001nM～1000nM投与してもよい。
The dosage of the ADC of the present invention necessary for treatment can vary depending on the age, sex, severity, etc. of the patient to be administered, but can ultimately be determined by the doctor in charge. For example, AD
C may be administered at a dose of 0.05-10 mg/kg body weight, preferably 0.1-2 mg/kg body weight. In addition, 0.001 nM to 1000 nM may be administered per dose in drug equivalents such as MMAE.
所定の投与量は１回の投与で与えてもよいし、１日当たり２回、３回、４回またはそれ
以上の分割投与とし、１日から数年にわたって、適当な間隔で投与してもよい。
The prescribed dose may be given in a single dose, or may be divided into two, three, four or more doses per day, administered at appropriate intervals over a period of one day to several years. .
本発明のADCを含む抗乳がん剤または抗胃がん剤は、他の抗がん剤や他の療法と併用す
ることもできる。他の療法として、例えば、放射線治療、ホルモン療法、外科手術等が挙
げられる。
The anti-breast cancer agent or anti-gastric cancer agent containing the ADC of the present invention can also be used in combination with other anti-cancer agents or other therapies. Other therapies include, for example, radiation therapy, hormone therapy, surgery, and the like.
本発明を以下の実施例によって具体的に説明するが、本発明はこれらの実施例によって
限定されるものではない。
The present invention will be specifically described by the following examples, but the invention is not limited by these examples.
[実施例１] 抗体-薬物複合体（ADC)の作製
抗体として、抗HER2抗体(トラスツズマブ：Trastuzumab）を使用した。トラスツズマブ
は、乳がん、胃がんに対する抗体医薬である。糖鎖部分の合成は以下のように行った。
[Example 1] Preparation of antibody-drug conjugate (ADC) An anti-HER2 antibody (trastuzumab) was used as an antibody. Trastuzumab is an antibody drug against breast cancer and gastric cancer. The sugar chain portion was synthesized as follows.
１． 薬物結合部位を有し、安定な構造を有する糖供与体の合成
DMF（333μL）中ジシアリルオクタサッカライド（30mg、0.0148mmol）、2-（2-（2-（2
-アジドエトキシ）エトキシ）エトキシ）エタン-1-アミン（DMF中4M、37μL、0.148mmol
）およびDIPEA（26μL、0.148mmol）の懸濁液に、PyBOP（77mg、0.148mmol）を4℃で加え
、混合物を40℃で8時間撹拌した。分析用HPLCは、0％ MeCNを含むC8逆相カラム（Mightys
il RP-8 GP、4.6×250mm、関東化学社）を用いて2分間、0.1％トリフルオロ酢酸水溶液中
0～100％MeCNの直線勾配で40分間行った（室温、流速1mL/分、214nm、280nmでの検出）。
8時間後、反応混合物をゲル濾過カラムクロマトグラフィー（G-25、GE Healthcare Life
Sciences、溶出液：H2O）により精製した。次いで、分取HPLC（Mightysil RP-18 GP、20
×250mm、関東化学）により、所望の生成物（アジド基を有する糖鎖）（20.4mg、収率57
％）を得た。
1H-NMR (D2O) d 5.18 (m, 1H), 5.10 (s, 1H), 4.91 (s, 1H), 4.57-4.55 (m, 2H), 4.42
-4.49 (m, 2H), 4.41-4.29 (m, 2H), 4.23 (s, 1H), 4.16 (s, 1H), 4.08 (s, 1H), 2.69
-2.65 (m, 2H), 2.02 (s, 15H), 2.00 (s, 9H), 1.87-1.78 (t, 2H); 13C-NMR (D2O) d 1
75.1, 174.8, 174.7, 169.2, 103.8, 103.8, 99.7, 99.5, 81.0, 80.9, 80.6, 80.4, 76.
5, 74.5, 76.3, 74.5, 73.7, 72.6, 74.5, 73.7, 72.6, 72.3, 72.2, 71.2, 70.8, 69.8,
69.6, 69.3, 69.3, 68.6, 68.4, 67.9, 67.5, 67.4, 67.1, 63.1, 60.3, 60.1, 54.8, 5
1.8, 50.3, 39.1, 38.9, 38.4, 38.4, 38.2, 38.0, 37.8, 37.6, 22.6, 22.2, 22.1; MS
calcd for [C92H152N13O61+Na+] 2443, found 2433.
1. Synthesis of glycosyl donors with drug-binding sites and stable structures
Disialyl octasaccharide (30 mg, 0.0148 mmol), 2-(2-(2-(2
-azidoethoxy)ethoxy)ethoxy)ethan-1-amine (4M in DMF, 37 μL, 0.148 mmol
) and DIPEA (26 μL, 0.148 mmol) was added PyBOP (77 mg, 0.148 mmol) at 4° C. and the mixture was stirred at 40° C. for 8 h. Analytical HPLC was run on a C8 reversed-phase column (Mightys
il RP-8 GP, 4.6×250mm, Kanto Kagaku) for 2 minutes in 0.1% trifluoroacetic acid aqueous solution
A linear gradient of 0-100% MeCN was run for 40 minutes (room temperature, flow
After 8 hours, the reaction mixture was subjected to gel filtration column chromatography (G-25, GE Healthcare Life
Sciences, eluent: H2O ). Preparative HPLC (Mightysil RP-18 GP, 20
× 250 mm, Kanto Kagaku), the desired product (sugar chain with an azide group) (20.4 mg, yield 57
%) was obtained.
1H - NMR ( D2O) d 5.18 (m, 1H), 5.10 (s, 1H), 4.91 (s, 1H), 4.57-4.55 (m, 2H), 4.42
-4.49 (m, 2H), 4.41-4.29 (m, 2H), 4.23 (s, 1H), 4.16 (s, 1H), 4.08 (s, 1H), 2.69
-2.65 (m, 2H), 2.02 (s, 15H), 2.00 (s, 9H), 1.87-1.78 (t, 2H); 13 C - NMR (D2O)
75.1, 174.8, 174.7, 169.2, 103.8, 103.8, 99.7, 99.5, 81.0, 80.9, 80.6, 80.4, 76.
5, 74.5, 76.3, 74.5, 73.7, 72.6, 74.5, 73.7, 72.6, 72.3, 72.2, 71.2, 70.8, 69.8,
69.6, 69.3, 69.3, 68.6, 68.4, 67.9, 67.5, 67.4, 67.1, 63.1, 60.3, 60.1, 54.8, 5
1.8, 50.3, 39.1, 38.9, 38.4, 38.4, 38.2, 38.0, 37.8, 37.6, 22.6, 22.2, 22.1; MS
calcd for [ C92H152N13O61 +Na + ] 2443 , found 2433 .
オキサゾリンの調製
D2O（590μL）中のラクトール（5.0mg、0.00207mmol）およびトリエチルアミン（20.2
μL、0.145mmol）の溶液に、塩化2-クロロ-1,3-ジメチル-1H-ベンズイミダゾール-3-イウ
ム（CDMBI、20.2 0.0930mmol）を4℃で添加し、2時間撹拌した。反応の進行を1H NMRによ
り分析し、オキサゾリンの形成を確認した。次いで、ゲル濾過カラムクロマトグラフィー
（G-25、GE Healthcare Life Sciences）、溶出液：0.1％トリエチルアミン水溶液）によ
り精製した。凍結乾燥後、所望の生成物（アジドオキサゾリン糖鎖）が74％の収率で得ら
れた。分析用および分取用HPLCは、C18逆相カラム（YMC-Triart C18; YMC Co.、Inc. 4.6
×150mm）を用いて0％MeCNで2分間、0.1％トリエチルアミン水溶液中15～37.5％MeCNの直
線勾配で45分間行った（室温、流速0.7mL /分、214nm、254nmでの検出）。
1H-NMR d 6.06 (d, J = 7.3 Hz, 1H), 5.09 (s, 1H), 4.92 (s, 1H), 4.57-4.55 (m, 2H)
, 4.41-4.39 (m, 2H), 4.34 (m, 1H), 4.15-4.12 (m, 4H), 2.68-2.65 (m, 2H), 2.03 (s
, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.98 (s, 3H), 1.80 (t, J = 13.2 Hz, 2H); 13C-
NMR (D2O) d 181.6, 175.1, 174.8, 169.2, 168.7, 103.8, 101.5, 99.5, 81.0, 80.6, 7
8.1, 76.5, 76.1, 74.5, 73.6, 72.6, 72.2, 71.2, 71.0, 70.8, 70.4, 69.8, 69.7, 69.
4, 69.3, 68.6, 68.5, 67.9, 67.4, 67.1, 63.2, 62.8, 61.8, 60.3, 54.7, 51.9, 50.3,
49.0, 38.9, 38.4, 23.4, 22.6, 22.0, 20.2, 13.1, 12.5.
Preparation of oxazolines
Lactol (5.0 mg, 0.00207 mmol) and triethylamine (20.2 mg) in D2O (590 μL)
μL, 0.145 mmol) was added 2-chloro-1,3-dimethyl-1H-benzimidazol-3-ium chloride (CDMBI, 20.2 0.0930 mmol) at 4° C. and stirred for 2 h. The progress of the reaction was analyzed by 1 H NMR, confirming the formation of oxazoline. Then, it was purified by gel filtration column chromatography (G-25, GE Healthcare Life Sciences), eluent: 0.1% triethylamine aqueous solution). After lyophilization, the desired product (azidoxazoline sugar chain) was obtained in 74% yield. Analytical and preparative HPLC was run on a C18 reversed-phase column (YMC-Triart C18; YMC Co., Inc. 4.6
× 150 mm) with 0% MeCN for 2 minutes and a linear gradient of 15-37.5% MeCN in 0.1% triethylamine aqueous solution for 45 minutes (room temperature, flow rate 0.7 mL/min, 214 nm, detection at 254 nm).
1 H-NMR d 6.06 (d, J = 7.3 Hz, 1H), 5.09 (s, 1H), 4.92 (s, 1H), 4.57-4.55 (m, 2H)
, 4.41-4.39 (m, 2H), 4.34 (m, 1H), 4.15-4.12 (m, 4H), 2.68-2.65 (m, 2H), 2.03 (s
, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.98 (s, 3H), 1.80 (t, J = 13.2 Hz, 2H); 13 C-
NMR ( D2O)d 181.6, 175.1, 174.8, 169.2, 168.7, 103.8, 101.5, 99.5, 81.0, 80.6, 7
8.1, 76.5, 76.1, 74.5, 73.6, 72.6, 72.2, 71.2, 71.0, 70.8, 70.4, 69.8, 69.7, 69.
4, 69.3, 68.6, 68.5, 67.9, 67.4, 67.1, 63.2, 62.8, 61.8, 60.3, 54.7, 51.9, 50.3,
49.0, 38.9, 38.4, 23.4, 22.6, 22.0, 20.2, 13.1, 12.5.
２． 糖鎖除去と糖転移酵素による抗体への転移
（１）トラスツズマブの糖鎖の除去
50mMリン酸緩衝液(pH 7.5)3mLにトラスツズマブ製剤を60 mg溶解し、そこに2 mg/mLのE
ndoS (E.coliより精製, His tag付き)50 mMリン酸緩衝液(pH 7.5)溶液30μLを添加し、37
℃でインキュベーションを16時間行った。16時間後、トラスツズマブの糖鎖の切断をUPLC
(AQUITY UPLC H-Class, Waters)で確認した。分析条件を以下に示した(分析法（１）)。
表１に移動相Ａと移動相Ｂの濃度勾配の詳細を示す。
2. Glycan removal and transfer to antibody by glycosyltransferase (1) Removal of trastuzumab sugar chain
Dissolve 60 mg of the trastuzumab formulation in 3 mL of 50 mM phosphate buffer (pH 7.5), add 2 mg/mL E
Add 30 μL of ndoS (purified from E. coli, with His tag) in 50 mM phosphate buffer (pH 7.5),
°C for 16 hours. UPLC for trastuzumab glycolysis after 16 hours
(AQUITY UPLC H-Class, Waters). The analytical conditions are shown below (analytical method (1)).
Table 1 shows the details of the concentration gradients of mobile phase A and mobile phase B.
LCシステム: AQUITY UPLC H-Class
カラム: AQUITY UPLC Glycoprotein Amide 300 Å(2.1 × 150 mm)
サンプラー温度: 8℃
カラム温度: 80℃
流速: 0.4 mL/分
検出: 蛍光、Ex : 280 nm、Em : 320 nm
移動相A : 0.1% TFA・0.3%HFIP（水中）
移動相B : 0.1%TFA・0.3%HFIP（アセトニトリル中）
LC System: AQUITY UPLC H-Class
Column: AQUITY UPLC Glycoprotein Amide 300Å (2.1 x 150 mm)
Sampler temperature: 8℃
Column temperature: 80℃
Flow rate: 0.4 mL/min Detection: fluorescence, Ex: 280 nm, Em: 320 nm
Mobile phase A: 0.1% TFA, 0.3% HFIP (in water)
Mobile phase B: 0.1% TFA, 0.3% HFIP (in acetonitrile)
回収したフラクションをAmicon 10K (Amicon Ultra 10K, Merck)で限外ろ過し濃縮した
(3000 rpm, 4℃)。次の糖鎖付加反応の反応緩衝液(50 mMリン酸緩衝液(pH 6.5))に置換す
るため、Amicon10Kに残る濃縮液の体積の10倍量の50 mMリン酸緩衝液(pH 6.5)を加え、限
外ろ過による濃縮を行った。緩衝液の置換では置換緩衝液の添加→濃縮の操作を計3回行
った。濃縮操作後、糖鎖が除去されたトラスツズマブの溶液濃度を測定するため、280 nm
における紫外可視吸光度測定を行った。
Collected fractions were ultrafiltered and concentrated with Amicon 10K (Amicon Ultra 10K, Merck).
(3000 rpm, 4°C). To replace the reaction buffer (50 mM phosphate buffer (pH 6.5)) for the next glycosylation reaction, add 10 times the volume of the concentrated solution remaining in the Amicon 10K with 50 mM phosphate buffer (pH 6.5). In addition, concentration was performed by ultrafiltration. In the replacement of the buffer solution, the operation of adding the replacement buffer solution and then concentrating was performed a total of three times. After concentration operation, 280 nm
UV-visible absorbance measurements were performed at .
（２）トラスツズマブへのアジドオキサゾリン糖鎖の付加
アジドオキサゾリン糖鎖を50 mMリン酸緩衝液(pH 8.0)に溶解し、0.1 mg/mLとした(オ
キサゾリン骨格は酸性条件により損壊するため、反応溶液である50 mMリン酸緩衝液(pH 6
.5)ではなく、pH 8.0の緩衝液によりストックソリューション化した)。（１）で精製した
トラスツズマブ0.1 mgを50 mMリン酸緩衝液(pH 6.5)で容量を40μLとし、15.0 mg/mLのEn
doS(D233Q) 50 mMリン酸緩衝液(pH 6.5)を2.34μL添加した。この反応溶液にアジドオキ
サゾリン糖鎖のストックソリューション0.1 mg/μLを0.75μL添加し、30℃でインキュベ
ーションを行った。15分毎にUPLC(分析法（１）)により反応解析を行い、糖鎖が除去され
たトラスツズマブとオキサゾリンが一つ付加したトラスツズマブのピークが消失した段階
でアジドオキサゾリン糖鎖の添加を止め、（１）と同様の手順で精製・濃縮を行った。最
後の緩衝液置換では次のクリック反応で使用する50 mMリン酸緩衝液(pH 8.0)を使用し、
（１）と同様の操作を行った。抗体濃度は分析法（１）と同様の操作により算出した。
(2) Addition of azidoxazoline sugar chain to trastuzumab The azidoxazoline sugar chain was dissolved in 50 mM phosphate buffer (pH 8.0) to a concentration of 0.1 mg/mL (since the oxazoline skeleton is destroyed under acidic conditions, the reaction solution 50 mM phosphate buffer (pH 6
.5) but made into a stock solution with pH 8.0 buffer). 0.1 mg of trastuzumab purified in (1) was adjusted to 40 μL with 50 mM phosphate buffer (pH 6.5), and 15.0 mg/mL of En
2.34 μL of doS(D233Q) 50 mM phosphate buffer (pH 6.5) was added. 0.75 µL of 0.1 mg/µL of stock solution of azidoxazoline sugar chain was added to this reaction solution and incubated at 30°C. Reaction analysis was performed by UPLC (analytical method (1)) every 15 minutes, and when the peaks of trastuzumab with the sugar chain removed and trastuzumab with one oxazoline added disappeared, the addition of the azidoxazoline sugar chain was stopped, and ( Purification and concentration were performed in the same procedure as in 1). The final buffer replacement uses the 50 mM phosphate buffer (pH 8.0) used in the next click reaction,
The same operation as in (1) was performed. Antibody concentrations were calculated in the same manner as in analytical method (1).
３．薬物の設計と合成と結合
Monomethyl auristatin E (MMAE)は、ADCに用いられる強力な活性を持つ抗がん剤であ
る。細胞内において抗体から放出可能であるように、カテプシン切断型のリンカーを用い
た。アジド基と選択的に反応可能であるひずみのかかったアルキンを反応させた。
反応式を化学式１に示す。
3. Drug design, synthesis and conjugation
Monomethyl auristatin E (MMAE) is a potent anticancer agent used in ADCs. A cathepsin-cleavable linker was used so that it could be released from the antibody intracellularly. A strained alkyne that is selectively reactive with the azide group was reacted.
The reaction formula is shown in
Ｂの合成
DMF（4.7mL）中のFmoc-Val-Cit-PABC-MMAE（0.34g）の溶液に、Et2NH（1.2mL）を添加
した。1時間後、混合物を減圧下で濃縮し、残渣をLH20（CHCl3：MeOH 1：1）で精製し、0
.24gのH-Val-Cit-PABC-MMAEを得た。CH2Cl2（10mL）中のH-Val-Cit-PABC-MMAE（0.234mmo
l）およびFmoc-PEG11-CO2H（0.19g、0.234mmol）の溶液に、WSCDI（67mg、0.351mmol）、
HOBt（47mg、0.351mmol）およびi-Pr2NEt（0.13mL、0.702mmol）を4℃で添加した。一晩
経過後、混合物をLH20（CHCl3：MeOH 1：1）を用いて精製して、0.31gの化合物Bを得た。
Synthesis of B
To a solution of Fmoc-Val-Cit-PABC - MMAE (0.34 g) in DMF (4.7 mL) was added Et2NH (1.2 mL). After 1 hour, the mixture was concentrated under reduced pressure and the residue was purified with LH20 (CHCl 3 :MeOH 1:1) and
.24 g of H-Val-Cit-PABC-MMAE was obtained. H - Val-Cit-PABC-MMAE (0.234 mmol) in
l) and to a solution of Fmoc-PEG11 - CO2H (0.19 g, 0.234 mmol), WSCDI (67 mg, 0.351 mmol),
HOBt (47 mg, 0.351 mmol) and i - Pr2NEt (0.13 mL, 0.702 mmol) were added at 4°C. After overnight, the mixture was purified using LH20 (CHCl 3 :MeOH 1:1) to give 0.31 g of compound B.
Ｃの合成
DMF（1.5mL）中のFmoc-PEG11-Val-Cit-PABC-MMAE（0.31g、0.160mmol）の溶液に、Et2N
H（0.3mL）を添加した。1時間後、混合物を減圧下で濃縮し、残渣をLH20（CHCl3：MeOH 1
：1）で精製して、0.27gのH-PEG11-Val-Cit-PABC-MMAEを得た。CH2Cl2（10mL）中のH-PEG
11-Val-Cit-PABC-MMAE（0.27g）およびDBCO酸（58mg、0.234mmol）の溶液に、WSCDI（46m
g、0.240mmol）、HOBt 、0.240mmol）およびi-Pr2NEt（83μL、0480mmol）を4℃で添加し
た。一晩経過後、混合物をLH20（CHCl3：MeOH 1：1）を用いて精製して、0.29gの化合物C
を得た。
Synthesis of C
To a solution of Fmoc-PEG 11 -Val-Cit-PABC-MMAE (0.31 g, 0.160 mmol) in DMF (1.5 mL) was added Et 2 N
H (0.3 mL) was added. After 1 hour, the mixture was concentrated under reduced pressure and the residue was treated with LH20 (CHCl 3 :
: 1) to give 0.27 g of H-PEG 11 -Val-Cit-PABC-MMAE. H - PEG in
To a solution of 11 -Val-Cit-PABC-MMAE (0.27 g) and DBCO acid (58 mg, 0.234 mmol), WSCDI (46 m
g, 0.240 mmol), HOBt, 0.240 mmol) and i - Pr2NEt (83 μL, 0480 mmol) were added at 4°C. After overnight, the mixture was purified using LH20 (CHCl3:MeOH 1 :1) to give 0.29 g of compound C.
got
薬物部位MMAE-PEG12の導入によるADCの作製
精製したアジドオキサゾリン糖鎖が付加されたトラスツズマブを最終濃度が1.78 mg/mL
になるように50 mMリン酸緩衝液(pH 8.0)に溶解し、25.3 mg/mLのMMAE-PEG12 (上記構造)
DMSO溶液を20当量分添加した。MMAE-PEG12の水溶性が低いため、反応溶液体積の30%にな
るようにDMSOを更に添加し、25℃でインキュベーションを16時間行った。16時間後、反応
溶液をmilliQで3倍希釈し、希釈液を上記２の分析法（１）と同様の手順で精製・濃縮を
行った。その後、上記２の分析法（１）と同様の操作により50 mMリン酸緩衝液(pH 7.5)
に置換し保存した。抗体濃度は分析法（１）と同様の操作により算出した。
Preparation of ADC by introduction of the drug moiety MMAE-PEG12 Purified azidoxazoline-glycosylated trastuzumab at a final concentration of 1.78 mg/mL
25.3 mg/mL MMAE-PEG12 (above structure) dissolved in 50 mM phosphate buffer (pH 8.0) to give
DMSO solution was added in 20 equivalents. Due to the low water solubility of MMAE-PEG 12 , additional DMSO was added to 30% of the reaction solution volume and incubated at 25°C for 16 hours. After 16 hours, the reaction solution was diluted 3-fold with milliQ, and the diluted solution was purified and concentrated in the same manner as the analysis method (1) in 2 above. After that, 50 mM phosphate buffer (pH 7.5) by the same operation as the analysis method (1) in 2 above
and saved. Antibody concentrations were calculated in the same manner as in analytical method (1).
構造の解析
構造はAspN（エンドプロテイナーゼ）やイムノグロブリン分解酵素により作製したADC
を分解し、質量分析により構造と均一性を確認した。
Structural analysis The structure is ADC prepared by AspN (endoproteinase) and immunoglobulin degrading enzyme
was resolved and the structure and homogeneity were confirmed by mass spectrometry.
[実施例２] ADCの活性測定
HER2を発現している細胞（MKN-45（胃がん（腺がん）細胞）、MCF-7（乳がん細胞）、N
-87（胃がん（管状腺がん）細胞）、OE-19（食道がん、胃がん細胞）およびSK-BR-3（乳
がん細胞）)に対する作製したトラスツズマブとMMAEのADCとトラスツズマブの結合を比較
した。MKN-45はHER2の発現が低い胃がん細胞、N-87、OE-19はHER2の発現が高い胃がん細
胞、MCF-7はHER2の発現が低い乳がん細胞、SK-BR-3はHER2の発現が低い乳がん細胞である
。
[Example 2] ADC activity measurement
HER2-expressing cells (MKN-45 (gastric cancer (adenocarcinoma) cells), MCF-7 (breast cancer cells), N
-87 (gastric cancer (tubular adenocarcinoma) cells), OE-19 (esophageal and gastric cancer cells) and SK-BR-3 (breast cancer cells)) compared the binding of engineered trastuzumab and MMAE ADCs to trastuzumab . MKN-45 is gastric cancer cells with low HER2 expression, N-87 and OE-19 are gastric cancer cells with high HER2 expression, MCF-7 is breast cancer cells with low HER2 expression, and SK-BR-3 is high HER2 expression. Low breast cancer cells.
また、トラスツズマブとMMAEのADCの殺細胞効果をトラスツズマブ単独およびMMAE単独
と比較した。
We also compared the ADC cell-killing effects of trastuzumab and MMAE with trastuzumab alone and MMAE alone.
１．方法
細胞培養
SK-BR-3、MCF-7およびNCI-N87 (N87)細胞はAmerican Type Culture Collection (ATCC)
から購入した。OE-19細胞は、European Collection of Cell Culture (ECACC)から入手し
た。MKN-45細胞はJapanese Collection of Research Bioresources (JCRB)から入手した
。OE-19およびNCI-N87細胞はRoswell Park Memorial Institute 培地 (RPMI) 1640により
維持した。MKN-45およびMCF-7細胞はEagle's minimal essential medium (EMEM)により維
持した。SK-BR-3細胞はMcCoy’s 5A培地により維持した。全ての培地は10％ウシ胎仔血清
(FBS)と1％の抗生物質を添加し、5％CO2下37℃にて培養した。
1. Method cell culture
SK-BR-3, MCF-7 and NCI-N87 (N87) cells are from the American Type Culture Collection (ATCC)
purchased from OE-19 cells were obtained from the European Collection of Cell Culture (ECACC). MKN-45 cells were obtained from the Japanese Collection of Research Bioresources (JCRB). OE-19 and NCI-N87 cells were maintained in Roswell Park Memorial Institute medium (RPMI) 1640. MKN-45 and MCF-7 cells were maintained in Eagle's minimal essential medium (EMEM). SK-BR-3 cells were maintained in McCoy's 5A medium. All media are 10% fetal bovine serum
(FBS) and 1% antibiotics were added and cultured at 37°C under 5% CO2 .
in vitro 細胞障害アッセイ
細胞を96穴プレート(Corning, USA)に5000個/ウェルとなるように分注し、10％FBSを含
む至適培地の中で24時間プレインキュベーションした。プレインキュベーション後、各細
胞に、MMAE、トラスツズマブあるいは、ADCを0.50 nM MMAE 等量となるように（トラスツ
ズマブはADCとタンパク質量が等しくなるように）各濃度で添加し、72時間培養した。培
養後、細胞の生存率をCell Counting Kit-8 (Dojindo Molecular Technologies, Japan)
により評価した。相対的な細胞生存率は、各ウェルの吸光度を、各プレートの薬剤を含ま
ない培地で処理したウェルの平均吸光度で割ることにより算出した。
In vitro Cytotoxicity Assay Cells were dispensed into 96-well plates (Corning, USA) at 5000 cells/well and pre-incubated for 24 hours in optimal medium containing 10% FBS. After preincubation, each cell was added with MMAE, trastuzumab, or ADC at a concentration equivalent to 0.50 nM MMAE (trastuzumab has the same protein amount as ADC) and cultured for 72 hours. After culturing, cell viability was measured using Cell Counting Kit-8 (Dojindo Molecular Technologies, Japan).
Evaluated by Relative cell viability was calculated by dividing the absorbance of each well by the average absorbance of drug-free medium-treated wells on each plate.
フローサイトメトリー
トラスツズマブまたはADCのHER2高発現細胞と低発現細胞への結合能についてはフロー
サイトメトリーにより以下のように評価した。細胞を、30mMクエン酸三ナトリウム二水和
物および270mM塩化カリウムを含有するリン酸緩衝生理食塩水（PBS）中において、37℃で
30分間処理し剥離させた。次いで、細胞を、2mMのEDTAおよび0.5％のウシ血清アルブミン
を含むPBSでブロッキングした。トラスツズマブおよびADCを10μg/mLで添加し、細胞を4
℃で1時間インキュベートし、続いて10μg/mLのGoat anti human-Alexa Fluor 647（Life
Technologies、USA）と共に4℃で1時間インキュベートした。試料は、Guava easyCyte T
M（Millipore、USA）を用いて分析した。データ解析はFlowJo 7.6.5 (FLOWJO, USA)によ
って実施した。
Flow cytometry The ability of trastuzumab or ADC to bind to HER2 high and low HER2 expressing cells was evaluated by flow cytometry as follows. Cells were incubated in phosphate buffered saline (PBS) containing 30 mM trisodium citrate dihydrate and 270 mM potassium chloride at 37°C.
Treated for 30 minutes and stripped. Cells were then blocked with PBS containing 2 mM EDTA and 0.5% bovine serum albumin. Trastuzumab and ADC were added at 10 μg/mL and the cells were
°C for 1 hour, followed by 10 μg/mL Goat anti human-Alexa Fluor 647 (Life
Technologies, USA) at 4°C for 1 hour. The sample was Guava easyCyte T
M (Millipore, USA) was used for analysis. Data analysis was performed by FlowJo 7.6.5 (FLOWJO, USA).
２．結果
結合能の確認
薬物の結合が抗体の抗原への結合に影響を及ぼさないことを確認するためにHER2低発現
と高発現細胞それぞれへの抗体またはADCの結合をフローサイトメトリーと用いて評価し
た。
2. Results Confirmation of binding ability Antibody or ADC binding to HER2 low- and high-expressing cells, respectively, was evaluated using flow cytometry to confirm that drug binding did not affect antigen binding of the antibody. .
結果を図３に示す。図３中「Trastuzumab-ADC」は、トラスツズマブとMMAEのADCを、「
Trastuzumab」はトラスツズマブ単独を、「2ndary only」はGoat anti human-Alexa Fluo
r 647のみの結果を示す。
The results are shown in FIG. In Figure 3, "Trastuzumab-ADC" is the ADC of trastuzumab and MMAE
Trastuzumab” is trastuzumab alone, and “Secondary only” is Goat anti human-Alexa Fluo
Results for r647 only are shown.
すべてのHER2低発現と高発現細胞それぞれにおいてトラスツズマブとADCは同様の結合
能を示した。これらの結果は、糖鎖への薬物の付加が抗原への結合に影響しないことを示
している。
Trastuzumab and ADC showed similar binding capacities in all HER2 low- and high-expressing cells, respectively. These results indicate that the addition of drugs to carbohydrate chains does not affect antigen binding.
in vitro細胞障害アッセイ
細胞のHER2発現とADCの細胞障害効果が正の相関があるかを評価するために、トラスツ
ズマブとADCとMMAEのHER2低発現細胞と高発現細胞それぞれに対しての細胞障害効果をin
vitroで測定した。
In vitro cytotoxicity assay To evaluate whether there is a positive correlation between HER2 expression in cells and the cytotoxic effect of ADC, the cytotoxic effects of trastuzumab, ADC, and MMAE on HER2-low and high-expressing cells, respectively in
measured in vitro.
図４～６に結果を示す。図４はNKN-45（HER2低発現）およびN-87（HER2高発現）に対す
る殺細胞効果を、図５はMKN-45（HER2低発現）およびOE-19（HER2高発現）に対する殺細
胞効果を、図６はMCF-7（HER2低発現）およびSK-BR-3（HER2高発現）に対する殺細胞効果
を示す。「MMAE」はMMAE単独の結果を、「Trastuzumab」はトラスツズマブ単独の結果を
、「Trastuzumab-ADC」はトラスツズマブとMMAEのADCの結果を示す。縦軸はMMAE濃度のと
きの細胞生存率を１とした場合の相対的な細胞生存率を示し、横軸はMMAEの量（nM 当量)
を示す。
The results are shown in Figures 4-6. Figure 4 shows the cell-killing effect on NKN-45 (low HER2 expression) and N-87 (high HER2 expression), and Figure 5 shows the cell-killing effect on MKN-45 (low HER2 expression) and OE-19 (high HER2 expression). and Figure 6 shows the cell-killing effect on MCF-7 (low HER2 expression) and SK-BR-3 (high HER2 expression). "MMAE" indicates results for MMAE alone, "Trastuzumab" indicates results for trastuzumab alone, and "Trastuzumab-ADC" indicates ADC results for trastuzumab and MMAE. The vertical axis shows the relative cell viability when the cell viability at the MMAE concentration is 1, and the horizontal axis shows the amount of MMAE (nM equivalent).
indicates
HER2発現が低い乳がん細胞であるMCF-7や胃がん細胞のMKN-45は抗体トラスツズマブに
対しても、ADCに対してもHER2抗原が少ないため効果を示さなかった。HER2発現が高い乳
がん細胞SK-BR-3や胃がん細胞OE-19やN87細胞に対しては、トラスツズマブ単独による弱
い細胞障害活性は、観察されたが、このアッセイ系では、効果が弱いため、IC50は決定で
きなかった（20μg/mL IgGまで）。一方で、HER2高発現細胞のADC への曝露は、細胞増殖
の用量依存的な減少を引き起こし、SK-BR-3、OE-19およびN87に対するIC50値は、それぞ
れ、0.3, 0.9, および、1.8 nM MMAE 当量 (MMAE eq.)であった。
これらの結果は、ADCの障害活性がHER2発現に依存するものであることを示している。
MCF-7, a breast cancer cell with low HER2 expression, and MKN-45, a gastric cancer cell, did not show any effect on the antibody trastuzumab or ADC due to their low HER2 antigen content. Weak cytotoxic activity by trastuzumab alone was observed against breast cancer cells SK-BR-3 and gastric cancer cells OE-19 and N87 cells with high HER2 expression. could not be determined (up to 20 μg/mL IgG). On the other hand, exposure of HER2-high expressing cells to ADC caused a dose-dependent decrease in cell proliferation, with IC50 values for SK-BR-3, OE-19 and N87 of 0.3, 0.9, and 1.8, respectively. nM MMAE equivalents (MMAE eq.).
These results indicate that the inhibitory activity of ADC is dependent on HER2 expression.
乳がん細胞であるSK-BR-3と胃がん細胞であるN-87はどちらもHER2を高発現している（
図３）。しかしながら、図４および図６を比較すると、SK-BR-3においては、ADCはMMAE単
独と同程度の殺細胞効果を示すが、N-87においては、ADCの効果はMMAE単独の効果よりも
小さい。本発明のADCは、細胞に到達し取り込まれるステップ、細胞に取り込まれた後に
薬物が放出されるステップの２つのステップを経て抗がん効果を発揮するので、２つのス
テップの効率がいずれも高い必要がある。図３の結果は、ADCはSK-BR-3およびN-87のいず
れにも高い効率で結合することを示している。また、カテプシンは細胞内に通常存在する
分解酵素であり、普遍的に作用する。従って、本発明のADCは乳がんまたは胃がんの治療
に特に有用である。
SK-BR-3, a breast cancer cell, and N-87, a gastric cancer cell, both highly express HER2 (
Figure 3). However, when comparing Figures 4 and 6, in SK-BR-3, ADC shows a similar cell-killing effect as MMAE alone, but in N-87, the effect of ADC is greater than that of MMAE alone. small. The ADC of the present invention exerts its anticancer effect through two steps: a step of reaching and being taken up by cells, and a step of being taken up by cells and then releasing the drug, so the efficiency of both steps is high. There is a need. The results in Figure 3 show that ADC binds both SK-BR-3 and N-87 with high efficiency. Moreover, cathepsins are degradative enzymes normally present in cells and act ubiquitously. Therefore, the ADCs of the invention are particularly useful for treating breast cancer or gastric cancer.
抗HER2抗体に抗がん剤を糖鎖およびカテプシン切断型リンカーを介して付加したADCは
、乳がんまたは胃がん治療に利用することができる。
ADCs, which are anti-HER2 antibodies attached with anticancer drugs via sugar chains and cathepsin-cleavable linkers, can be used for the treatment of breast cancer or gastric cancer.
Claims (7)
ペプチドリンカーおよび糖鎖を介して付加されており、細胞中でカテプシンによりリンカ
ーが切断され、抗がん剤が放出される、抗体-薬物複合体。 Valine-citrulline (Val-Cit), which is an anti-HER2 antibody and an anticancer drug, is a cathepsin-cleavable linker
An antibody-drug conjugate that is attached via a peptide linker and a sugar chain, in which the linker is cleaved by cathepsins in the cell, releasing the anticancer drug.
D)およびmonomethyl auristatin F (MMAF)からなる群から選択される、請求項２に記載の
抗体-薬物複合体。 Auristatin is monomethyl auristatin E (MMAE), monomethyl auristatin D (MMA
D) and monomethyl auristatin F (MMAF).
１項に記載の抗体-薬物複合体。 The antibody-drug conjugate according to any one of claims 1 to 3, wherein the anti-HER2 antibody is a sugar chain-engineered antibody having a uniform sugar chain structure.
。 The antibody-drug conjugate of any one of claims 1-4, wherein the antibody is trastuzumab.
療剤。 A therapeutic agent for breast cancer or gastric cancer, comprising the antibody-drug conjugate according to any one of claims 1 to 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022162603A JP2022176381A (en) | 2018-05-29 | 2022-10-07 | Antibody-drug conjugates for treating breast cancer or gastric cancer |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2018102760A JP2019206491A (en) | 2018-05-29 | 2018-05-29 | Antibody-drug complex for treatment of breast or gastric cancer |
| JP2022162603A JP2022176381A (en) | 2018-05-29 | 2022-10-07 | Antibody-drug conjugates for treating breast cancer or gastric cancer |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2018102760A Division JP2019206491A (en) | 2018-05-29 | 2018-05-29 | Antibody-drug complex for treatment of breast or gastric cancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2022176381A true JP2022176381A (en) | 2022-11-25 |
Family
ID=68768036
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2018102760A Pending JP2019206491A (en) | 2018-05-29 | 2018-05-29 | Antibody-drug complex for treatment of breast or gastric cancer |
| JP2022162603A Pending JP2022176381A (en) | 2018-05-29 | 2022-10-07 | Antibody-drug conjugates for treating breast cancer or gastric cancer |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2018102760A Pending JP2019206491A (en) | 2018-05-29 | 2018-05-29 | Antibody-drug complex for treatment of breast or gastric cancer |
Country Status (1)
| Country | Link |
|---|---|
| JP (2) | JP2019206491A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPWO2022211075A1 (en) * | 2021-03-31 | 2022-10-06 |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG187517A1 (en) * | 2008-01-31 | 2013-02-28 | Genentech Inc | Anti-cd79b antibodies and immunoconjugates and methods of use |
| BR112014030098A2 (en) * | 2012-06-04 | 2017-07-25 | Irm Llc | site-specific labeling methods and molecules produced by them |
| PL2911699T3 (en) * | 2012-10-23 | 2018-04-30 | Synaffix B.V. | Modified antibody, antibody-conjugate and process for the preparation thereof |
| EP2935611B1 (en) * | 2013-10-14 | 2021-07-07 | SynAffix B.V. | Glycoengineered antibody, antibody-conjugate and methods for their preparation |
| JP2017200902A (en) * | 2016-02-08 | 2017-11-09 | シンアフィックス ビー．ブイ． | Novel antibody-conjugate with improved therapeutic index for targeting HER2 tumor and method for improving therapeutic index of antibody-conjugate |
-
2018
- 2018-05-29 JP JP2018102760A patent/JP2019206491A/en active Pending
-
2022
- 2022-10-07 JP JP2022162603A patent/JP2022176381A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| JP2019206491A (en) | 2019-12-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7257422B2 (en) | Anti-MUC1 antibody-drug conjugate | |
| US20240173427A1 (en) | Glypican 3 antibodies and conjugates thereof | |
| JP2019142870A (en) | Hydrazinyl-pyrrolo compounds and methods for producing conjugate | |
| KR20210038904A (en) | Effective method for preparing antibody-drug conjugates | |
| US20220177601A1 (en) | Anti-her2 antibody-pyrrolobenzodiazepine derivative conjugate | |
| KR20160113623A (en) | Antibody-drug conjugates and immunotoxins | |
| KR20160113620A (en) | Antibody-drug conjugates and immunotoxins | |
| US20210061916A1 (en) | Anti-prlr antibody-drug conjugates (adc) and uses thereof | |
| JP2020143084A (en) | Antibody-urease conjugates for therapeutic purposes | |
| AU2015243512A1 (en) | Site-specific antibody-drug glycoconjugates and methods | |
| EP1098666B1 (en) | Water-soluble drugs and methods for their production | |
| JP2022176381A (en) | Antibody-drug conjugates for treating breast cancer or gastric cancer | |
| US7026350B2 (en) | Water-soluble drugs and methods for their production | |
| EP4392078A2 (en) | Methods of using antibody-drug-conjugates | |
| AU2022320714A1 (en) | Antibody-drug conjugates and methods of use thereof | |
| US11879004B2 (en) | Modified binding polypeptides for optimized drug conjugation | |
| US20230338568A1 (en) | Novel isomeric compounds comprising a ring-opened thiosuccinimide group, an oligopeptide fragment and a chiral moiety | |
| KR102110182B1 (en) | Novel oxime derivative compounds and antibody-drug conjugate comprising thereof | |
| WO2023053082A1 (en) | Lectin-drug conjugates | |
| JP2022552349A (en) | B-lymphocyte-specific amatoxin antibody conjugates | |
| KR20240095532A (en) | Lectin-drug conjugates | |
| JP2023551203A (en) | Glycosidic double cleavage linkers for antibody-drug conjugates | |
| CN116615256A (en) | Glycoconjugates |