CN111148985B - 评估抗体-药物缀合物的方法 - Google Patents
评估抗体-药物缀合物的方法 Download PDFInfo
- Publication number
- CN111148985B CN111148985B CN201880058051.6A CN201880058051A CN111148985B CN 111148985 B CN111148985 B CN 111148985B CN 201880058051 A CN201880058051 A CN 201880058051A CN 111148985 B CN111148985 B CN 111148985B
- Authority
- CN
- China
- Prior art keywords
- antibody
- sample
- drug
- light wavelength
- absorbance
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 60
- 229940049595 antibody-drug conjugate Drugs 0.000 title claims description 65
- 239000000611 antibody drug conjugate Substances 0.000 title claims description 41
- 229940079593 drug Drugs 0.000 claims description 51
- 239000003814 drug Substances 0.000 claims description 51
- 238000002835 absorbance Methods 0.000 claims description 46
- 238000001542 size-exclusion chromatography Methods 0.000 claims description 31
- 230000008033 biological extinction Effects 0.000 claims description 16
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- 239000012504 chromatography matrix Substances 0.000 claims description 6
- 229940127089 cytotoxic agent Drugs 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 230000000259 anti-tumor effect Effects 0.000 claims description 3
- 239000000562 conjugate Substances 0.000 claims description 3
- 239000000824 cytostatic agent Substances 0.000 claims description 3
- 239000002254 cytotoxic agent Substances 0.000 claims description 3
- 231100000599 cytotoxic agent Toxicity 0.000 claims description 3
- 230000000861 pro-apoptotic effect Effects 0.000 claims description 3
- 238000001195 ultra high performance liquid chromatography Methods 0.000 claims 2
- 238000011156 evaluation Methods 0.000 abstract description 3
- 239000012895 dilution Substances 0.000 abstract description 2
- 238000010790 dilution Methods 0.000 abstract description 2
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 21
- 239000000523 sample Substances 0.000 description 18
- 238000009472 formulation Methods 0.000 description 14
- 239000000203 mixture Substances 0.000 description 14
- 239000008186 active pharmaceutical agent Substances 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 238000004611 spectroscopical analysis Methods 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 4
- 229940088679 drug related substance Drugs 0.000 description 4
- 230000010354 integration Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 229930126263 Maytansine Natural products 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 239000002246 antineoplastic agent Substances 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 229960004679 doxorubicin Drugs 0.000 description 3
- 239000013583 drug formulation Substances 0.000 description 3
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 description 3
- 239000012898 sample dilution Substances 0.000 description 3
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 3
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 108010044540 auristatin Proteins 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229930013356 epothilone Natural products 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 229940121372 histone deacetylase inhibitor Drugs 0.000 description 2
- 239000003276 histone deacetylase inhibitor Substances 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 231100000647 material safety data sheet Toxicity 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- -1 melitracin Chemical compound 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- VSNHCAURESNICA-NJFSPNSNSA-N 1-oxidanylurea Chemical compound N[14C](=O)NO VSNHCAURESNICA-NJFSPNSNSA-N 0.000 description 1
- NDMPLJNOPCLANR-UHFFFAOYSA-N 3,4-dihydroxy-15-(4-hydroxy-18-methoxycarbonyl-5,18-seco-ibogamin-18-yl)-16-methoxy-1-methyl-6,7-didehydro-aspidospermidine-3-carboxylic acid methyl ester Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 NDMPLJNOPCLANR-UHFFFAOYSA-N 0.000 description 1
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 description 1
- NMUSYJAQQFHJEW-KVTDHHQDSA-N 5-azacytidine Chemical compound O=C1N=C(N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 NMUSYJAQQFHJEW-KVTDHHQDSA-N 0.000 description 1
- WYWHKKSPHMUBEB-UHFFFAOYSA-N 6-Mercaptoguanine Natural products N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 1
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 108090000323 DNA Topoisomerases Proteins 0.000 description 1
- 102000003915 DNA Topoisomerases Human genes 0.000 description 1
- 229940123780 DNA topoisomerase I inhibitor Drugs 0.000 description 1
- 229940124087 DNA topoisomerase II inhibitor Drugs 0.000 description 1
- 108010092160 Dactinomycin Proteins 0.000 description 1
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 102000003964 Histone deacetylase Human genes 0.000 description 1
- 108090000353 Histone deacetylase Proteins 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-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
- 239000005517 L01XE01 - Imatinib Substances 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- 239000000365 Topoisomerase I Inhibitor Substances 0.000 description 1
- 239000000317 Topoisomerase II Inhibitor Substances 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
- 229940122803 Vinca alkaloid Drugs 0.000 description 1
- IEDXPSOJFSVCKU-HOKPPMCLSA-N [4-[[(2S)-5-(carbamoylamino)-2-[[(2S)-2-[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]-3-methylbutanoyl]amino]pentanoyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate Chemical compound CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C)[C@@H](O)c1ccccc1)OC)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C(=O)OCc1ccc(NC(=O)[C@H](CCCNC(N)=O)NC(=O)[C@@H](NC(=O)CCCCCN2C(=O)CCC2=O)C(C)C)cc1)C(C)C IEDXPSOJFSVCKU-HOKPPMCLSA-N 0.000 description 1
- 229930183665 actinomycin Natural products 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 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
- 239000012491 analyte Substances 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 229960002756 azacitidine Drugs 0.000 description 1
- 229960002170 azathioprine Drugs 0.000 description 1
- LMEKQMALGUDUQG-UHFFFAOYSA-N azathioprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC=NC2=C1NC=N2 LMEKQMALGUDUQG-UHFFFAOYSA-N 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
- GXJABQQUPOEUTA-RDJZCZTQSA-N bortezomib Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)B(O)O)NC(=O)C=1N=CC=NC=1)C1=CC=CC=C1 GXJABQQUPOEUTA-RDJZCZTQSA-N 0.000 description 1
- 229960001467 bortezomib Drugs 0.000 description 1
- 229960004117 capecitabine Drugs 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 190000008236 carboplatin Chemical compound 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 1
- 229960004630 chlorambucil Drugs 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229940109262 curcumin Drugs 0.000 description 1
- 235000012754 curcumin Nutrition 0.000 description 1
- 239000004148 curcumin Substances 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 229960000975 daunorubicin Drugs 0.000 description 1
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 239000002552 dosage form Substances 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
- 238000007876 drug discovery Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229960001904 epirubicin Drugs 0.000 description 1
- HESCAJZNRMSMJG-KKQRBIROSA-N epothilone A Chemical class C/C([C@@H]1C[C@@H]2O[C@@H]2CCC[C@@H]([C@@H]([C@@H](C)C(=O)C(C)(C)[C@@H](O)CC(=O)O1)O)C)=C\C1=CSC(C)=N1 HESCAJZNRMSMJG-KKQRBIROSA-N 0.000 description 1
- 150000003883 epothilone derivatives Chemical class 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
- 229960005420 etoposide Drugs 0.000 description 1
- 229960002949 fluorouracil Drugs 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
- 229960005277 gemcitabine Drugs 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 229960002411 imatinib Drugs 0.000 description 1
- KTUFNOKKBVMGRW-UHFFFAOYSA-N imatinib Chemical compound C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 KTUFNOKKBVMGRW-UHFFFAOYSA-N 0.000 description 1
- 229960004768 irinotecan Drugs 0.000 description 1
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 1
- 229940043355 kinase inhibitor Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 210000001006 meconium Anatomy 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
- 229960000485 methotrexate Drugs 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 229960001156 mitoxantrone Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- 229950007318 ozogamicin Drugs 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 239000003757 phosphotransferase inhibitor Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 229960004641 rituximab Drugs 0.000 description 1
- HNMATTJJEPZZMM-BPKVFSPJSA-N s-[(2r,3s,4s,6s)-6-[[(2r,3s,4s,5r,6r)-5-[(2s,4s,5s)-5-[acetyl(ethyl)amino]-4-methoxyoxan-2-yl]oxy-6-[[(2s,5z,9r,13e)-13-[2-[[4-[(2e)-2-[1-[4-(4-amino-4-oxobutoxy)phenyl]ethylidene]hydrazinyl]-2-methyl-4-oxobutan-2-yl]disulfanyl]ethylidene]-9-hydroxy-12-(m Chemical compound C1[C@H](OC)[C@@H](N(CC)C(C)=O)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@@](C/3=C/CSSC(C)(C)CC(=O)N\N=C(/C)C=3C=CC(OCCCC(N)=O)=CC=3)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HNMATTJJEPZZMM-BPKVFSPJSA-N 0.000 description 1
- 229950000143 sacituzumab govitecan Drugs 0.000 description 1
- ULRUOUDIQPERIJ-PQURJYPBSA-N sacituzumab govitecan Chemical compound N([C@@H](CCCCN)C(=O)NC1=CC=C(C=C1)COC(=O)O[C@]1(CC)C(=O)OCC2=C1C=C1N(C2=O)CC2=C(C3=CC(O)=CC=C3N=C21)CC)C(=O)COCC(=O)NCCOCCOCCOCCOCCOCCOCCOCCOCCN(N=N1)C=C1CNC(=O)C(CC1)CCC1CN1C(=O)CC(SC[C@H](N)C(O)=O)C1=O ULRUOUDIQPERIJ-PQURJYPBSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000003998 size exclusion chromatography high performance liquid chromatography Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000008685 targeting Effects 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
- NRUKOCRGYNPUPR-QBPJDGROSA-N teniposide 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](OC[C@H]4O3)C=3SC=CC=3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 NRUKOCRGYNPUPR-QBPJDGROSA-N 0.000 description 1
- 229960001278 teniposide Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229960003087 tioguanine Drugs 0.000 description 1
- MNRILEROXIRVNJ-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=NC=N[C]21 MNRILEROXIRVNJ-UHFFFAOYSA-N 0.000 description 1
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- 229960001612 trastuzumab emtansine Drugs 0.000 description 1
- 229960003048 vinblastine Drugs 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
- 229960004528 vincristine Drugs 0.000 description 1
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 1
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 1
- 229960004355 vindesine Drugs 0.000 description 1
- UGGWPQSBPIFKDZ-KOTLKJBCSA-N vindesine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(N)=O)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1N=C1[C]2C=CC=C1 UGGWPQSBPIFKDZ-KOTLKJBCSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/3103—Atomic absorption analysis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/34—Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/42—Absorption spectrometry; Double beam spectrometry; Flicker spectrometry; Reflection spectrometry
- G01J3/427—Dual wavelengths spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- G01N31/02—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using precipitation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
- G16B5/00—ICT specially adapted for modelling or simulations in systems biology, e.g. gene-regulatory networks, protein interaction networks or metabolic networks
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16C—COMPUTATIONAL CHEMISTRY; CHEMOINFORMATICS; COMPUTATIONAL MATERIALS SCIENCE
- G16C20/00—Chemoinformatics, i.e. ICT specially adapted for the handling of physicochemical or structural data of chemical particles, elements, compounds or mixtures
- G16C20/30—Prediction of properties of chemical compounds, compositions or mixtures
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N2021/3129—Determining multicomponents by multiwavelength light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Theoretical Computer Science (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Crystallography & Structural Chemistry (AREA)
- Evolutionary Biology (AREA)
- Medical Informatics (AREA)
- Physiology (AREA)
- Biophysics (AREA)
- Computing Systems (AREA)
- Mathematical Physics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本公开提供了评估ADC产品的DAR的方法,这些方法提供相对于已知方法的优势。具体地,本公开的方法可以在高通量的应用中和/或在评估期间无需稀释ADC样品的情况下使用。
Description
相关申请的交叉引用
本申请要求2017年9月8日提交的美国临时专利申请号62/556,153的优先权,该临时专利申请的内容通过引用并入本文。
背景技术
抗体-药物-缀合物(ADC)是一类新兴的药物分子。它们定位到特异性靶标并递送有效药物的能力使其成为开发基于靶标的治疗产品的有吸引力的选择。ADC是通过将有效药物分子与单克隆抗体化学接头来产生的。与单克隆抗体缀合的药物分子的平均数量称为药物与抗体之比(“DAR”)。DAR是ADC产品的重要质量属性,因为它会影响产品的功效、安全性和/或稳定性。因此,以可靠且高通量的方式评估ADC产品的DAR的方法是可取的。
具体实施方式
本公开提供了评估ADC产品的DAR的方法,这些方法提供相对于已知方法的优势。具体地,本公开的方法可以在高通量的应用中和/或在评估期间无需稀释ADC样品的情况下使用。
紫外-可见(UV-Vis)和比尔-朗伯(Beer-Lambert)定律
DAR传统上是使用UV-Vis光谱法测量的(参见例如,Chen,MethodsMol.Biol.1045:267-73(2013))。该分析的基础是比尔-朗伯定律,即物质的吸光度与浓度之间的正比关系:
A=εcl,
其中A是吸光度,ε是消光系数(物质的物理常数),l是穿过含有细胞的分析物的路径长度,并且c是浓度。
使用UV-Vis光谱法对ADC产品进行DAR测量依赖于抗体的最大吸收(例如,280nm)与药物的最大吸收(例如,252nm)之差。例如,平均DAR可以使用缀合材料在280nm和252nm处测量的吸收的差来计算。尽管UV-Vis方法已在工业中广泛使用,但它缺少进行配制筛选研究所需的通量。它在不进行样品稀释的情况下无法使用,从而导致与样品稀释有关的误差。
因此,本公开至少部分地基于使用尺寸排阻色谱法(例如,UPLC)和斜率光谱法测量DAR的替代方法。对这些方法进行了表征,并在再现性、精密度和灵敏度方面与UV-Vis光谱法进行了比较。生成的数据支持使用基于UPLC的DAR方法来克服传统UV-Vis方法的通量限制。此外,基于斜率光谱法的方法可用于在不进行样品稀释的情况下分析ADC样品。
基于UPLC的方法
在一个实施方案中,尺寸排阻用于确定DAR。在一些实施方案中,本文公开的方法包括将包含抗体-药物缀合物的样品施加于尺寸排阻色谱基质。在一些实施方案中,本文公开的方法包括施加包含抗体-药物缀合物的样品并通过尺寸排阻色谱基质运行包含抗体-药物缀合物的样品。在一些实施方案中,将ADC样品的总量施加于尺寸排阻基质以进行分析。例如,以下基于UPLC的方法用于评估DAR。
使用Empower的Apex Track积分方法以及峰肩检测对280nm处收集的数据进行积分。保留时间积分范围因分子而异,但通常在3-9分钟内。具有最大高度和面积的峰被归类为“天然”、“主”或“单体”峰。早于“天然”峰洗脱的任何峰均归类为“HMW”峰。晚于“天然峰”洗脱的任何峰均归类为“LMW”峰。
从各个峰的面积与所有峰的总面积之比计算每个种类的相对百分比。以下的相对百分比面积报告为纯度的指标:总HMW%、天然(或主或单体)天然%或总LMW%。对所有峰的总面积求和并用于随后的DAR计算中。然而,在一些实施方案中,仅使用天然峰的面积。
使用Empower的Apex Track积分方法以及峰肩检测对252nm处收集的数据进行了积分。保留时间积分范围因分子而异,但通常在3-9分钟内。对所有峰的总面积求和并用于随后的DAR计算中。然而,在一些实施方案中,仅使用天然峰的面积。
DAR是由280nm处的总峰面积(ADC的Amax)和252nm处的总峰面积(药物的Amax)来确定。尽管对于用于ADC的药物缀合物来说252nm是常见的Amax,但是可以例如使用已知方法为特定的缀合物选择适当的波长。如果适用,可以使用裸抗体作为参考标准品,通过这两个波长处的总峰面积之差来确定与抗体结合的药物的量。
以下两个公式(从比尔-朗伯定律推导出来)已被验证并证明为具有一致性。
公式1:
公式1不需要使用裸抗体参考标准品。然而,需要系统性确定抗体和药物二者在252nm处的消光系数(ε)。给定波长处的消光系数可以由比尔-朗伯定律通过使用已知浓度的抗体或药物的溶液并测量给定波长处的吸光度容易地计算。
公式2:
公式2不需要对252nm处的抗体进行消光系数确定,但确实需要收集裸抗体参考标准品的UPLC数据。
尽管已经举例说明了UPLC,但是在本文描述的方法中可以使用其他尺寸排阻色谱技术。尺寸排阻色谱法通常是指按尺寸分离分子,其中色谱洗脱时间对于特定分子是特征性的。另外的方法包括例如SEC-HPLC、反相(RP)HPLC、RP-UPLC。
在一些实施方案中,在通过尺寸排阻色谱法(例如HPLC或UPLC)进行分析之前,不稀释ADC样品。在一些实施方案中,在通过尺寸排阻色谱法进行分析之前ADC样品不需要进行稀释,因为将ADC样品的总量施加于尺寸排阻色谱基质。在一些实施方案中,分析含有约1μg/μL至约500μg/μLADC的样品。
基于斜率光谱法的方法
在一些实施方案中,DAR是通过计算ADC样品中抗体和药物的浓度来确定。例如,斜率光谱法是用于确定各种路径长度下溶液的吸光度的已知方法。然后可以基于比尔-朗伯定律使用各种路径长度下的吸光度值来计算溶液中化合物的浓度。采用斜率光谱法的方法和系统是已知的(参见例如,美国公开号20120130649)和可商购的(参见例如,SoloVPE(CTechnologies,Inc.,Bridgewater,NJ))。此类方法和系统适用于测量ADC制剂中抗体和药物的浓度,从而确定DAR。
例如,可以将ADC样品放置在容器中;可以使探针相对于容器移动,使得探针与容器的底部接触;可以使探针相对于容器移动,使得探针从容器的底部穿过样品以预定的增量移动,从而获得通过溶液的预先选择的路径长度;可以在抗体的最大吸收处读取吸光度读数;可以使探针相对于样品反复移动,并可以进行测量;可以从吸光度和路径长度生成回归线,从而获得回归线的斜率;并且可以通过将回归线的斜率除以抗体的消光系数来确定抗体的浓度。然后可以使用药物的最大吸收来重复这些步骤,以确定药物的浓度。DAR可以由确定的药物浓度和抗体浓度来计算。
在一些实施方案中,在通过斜率光谱法进行分析之前,不稀释ADC样品。在一些实施方案中,分析含有约0.1μg/μL至约500μg/μL ADC的样品。
抗体-药物缀合物
如本文所用,术语“抗体-药物缀合物”是指通过将抗体与生物活性细胞毒性有效载荷或药物连接而产生的蛋白质。抗体-药物缀合物(ADC)是通常通过本领域技术人员已知的化学修饰/偶联反应产生的。可以使用本文描述的方法分析任何抗体-药物缀合物。
在一些实施方案中,抗体-药物缀合物包括抗肿瘤抗体(参见例如,Adler等人,Hematol.Oncol.Clin.North Am.26:447-81(2012);Li等人,Drug Discov.Ther.7:178-84(2013);Scott等人,Cancer Immun.12:14(2012);和Sliwkowski等人,Science 341:1192-1198(2013))。表1展现了通过已知的、可用的抗体药剂靶向的某些人多肽抗原的非全面列表,并且记录了已被提出可以使用所述抗体药剂的某些癌症适应症。表1中的任何抗体都可以包含在使用本公开方法评估的抗体-药物缀合物中。
表1:
在一些实施方案中,抗体-药物缀合物包括作为促细胞凋亡剂、细胞生长抑制剂和/或细胞毒性剂中的一种或多种的药物,例如具体包括用于和/或推荐用于治疗一种或多种与不良细胞增殖有关的疾病、障碍或病症的药剂。在许多实施方案中,药物是可用于治疗癌症的化学治疗剂。在一些实施方案中,化学治疗剂可以是或包括一种或多种烷基化剂、一种或多种蒽环类、一种或多种细胞骨架破坏剂(例如,微管靶向剂,如紫杉烷、美坦辛(maytansine)及其类似物)、一种或多种埃博霉素、一种或多种组蛋白脱乙酰酶抑制剂(HDAC)、一种或多种拓扑异构酶抑制剂(例如,拓扑异构酶I和/或拓扑异构酶II抑制剂)、一种或多种激酶抑制剂、一种或多种核苷酸类似物或核苷酸前体类似物、一种或多种肽抗生素、一种或多种基于铂的药剂、一种或多种类维生素A、一种或多种长春花生物碱和/或以下一种或多种的一种或多种类似物(即,具有相关的抗增殖活性)。在一些特定的实施方案中,化学治疗剂可以是或包括以下各项中的一种或多种:放线菌素、全反式视黄酸、奥瑞斯他汀(Auiristatin)、阿扎胞苷、硫唑嘌呤、博来霉素、硼替佐米、卡铂、卡培他滨、顺铂、苯丁酸氮芥、环磷酰胺、姜黄素、阿糖胞苷、柔红霉素、多西他赛、去氧氟尿苷、多柔比星、表柔比星、埃博霉素、依托泊苷、氟尿嘧啶、吉西他滨、羟基脲、伊达比星、伊马替尼、伊立替康、美坦辛和/或其类似物(例如DM1)、氮芥、巯嘌呤、甲氨蝶呤、米托蒽醌、抗美登醇(Maytansinoid)、奥沙利铂、紫杉醇、培美曲塞、替尼泊苷、硫鸟嘌呤、托泊替康、戊柔比星、长春碱、长春新碱、长春地辛、长春瑞滨及其组合。
在一些实施方案中,使用本公开的方法评估的抗体-药物缀合物是hLL1-阿霉素、hRS7-SN-38、hMN-14-SN-38、hLL2-SN-38、hA20-SN-38、hPAM4-SN-38、hLL1-SN-38、hRS7-Pro-2-P-Dox、hMN-14-Pro-2-P-Dox、hLL2-Pro-2-P-Dox、hA20-Pro-2-P-Dox、hPAM4-Pro-2-P-Dox、hLL1-Pro-2-P-Dox、P4/D10-阿霉素、吉妥单抗、本妥昔单抗、曲妥珠单抗emtansine、奥英妥珠单抗ozogamicin、glembatumomab vedotin、SAR3419、SAR566658、BIIB015、BT062、CMC-544、SAR3419、CDX-011、SGN-75、SGN-CD19A、AMG-172、AMG-595、BAY-94-9343、ASG-5ME、ASG-22ME、ASG-16M8F、MDX-1203、MLN-0264、抗-PSMA ADC、RG-7450、RG-7458、RG-7593、RG-7596、RG-7598、RG-7599、RG-7600、RG-7636、ABT-414、IMGN-853、IMGN-529、IMGN-901、vorsetuzumab mafodotin或lorvotuzumab mertansine(参见例如、Sassoon等人,MethodsMol.Biol.1045:1-27(2013);Bouchard等人,Bioorganic Med.Chem.Lett.24:5357-5363(2014))。
应用
本公开的方法具有多种应用并且包括,例如在原料药或药物制剂制造的不同阶段的质量控制,在原料药或药物制剂制造完成之前和/或之后(例如,在分配到填充/结束环境或设备之前或之后)、在将原料药或药物制剂发布至市售之前或之后(例如,在分配给药房、护理者、患者或其他终端用户之前)ADC制剂的分析。在一些情况下,ADC制剂是原料药(活性药物成分或“API”)或药物制剂(配制用于受试者如人类患者的API)。在一些情况下,ADC制剂来自制造或使用阶段,所述制造或使用阶段在发布给护理者或其他终端用户之前;在包装成单个剂型(如注射器、笔、小瓶或多剂量小瓶)之前;在确定所述批次可以商业发布之前;在生产所述制剂的检验证书、材料安全数据表(MSDS)或检验报告(CofA)之前。
来自本文所述方法的评估可用于指导、控制或实现ADC制剂的制备、分发和监控过程中的许多活动或步骤,并提供安全且有效的ADC制剂使用。因此,在例如响应于评价的实施方案中,例如根据是否满足标准(例如,特定的DAR、平均DAR和/或DAR范围),做出决定或采取步骤。本文所述的方法可以包括做出以下决定:(a)关于是否可以将ADC制剂配制成原料药或药物制剂;(b)关于是否可以对ADC制剂进行再加工(例如,所述制剂可以重复先前的加工步骤);和/或(c)ADC制剂不适合配制成原料药或药物制剂。在一些情况下,方法包括:如步骤(a)中所述进行配制,如步骤(b)中所述进行再加工或使所述制剂无法用于商业发布,例如通过对其进行标记或销毁,如步骤(c)中所述。
Claims (18)
1.一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其包括:
将所述样品施加于尺寸排阻色谱基质;
检测所述样品在第一光波长λ1处的吸光度响应,其中所述第一光波长是所述抗体的预定吸光度最大值;
检测所述样品在第二光波长λ2处的吸光度响应,其中所述第二光波长是所述药物的预定吸光度最大值;
确定所述样品在所述第一光波长处的总吸光度和所述样品在所述第二光波长处的总吸光度,各个总吸光度通过经由对高分子量峰、低分子量峰和主峰的面积求和对在一定洗脱时间间隔内吸光度响应的峰进行积分来计算;以及,
使用以下公式1计算所述药物与抗体之比:
其中是所述抗体在所述第一光波长处的消光系数;/>是所述抗体在所述第二光波长处的消光系数;/>是所述药物在所述第一光波长处的消光系数;/>是所述药物在所述第二光波长处的消光系数;总面积λ1是所述样品在所述第一光波长处的总吸光度;并且总面积λ2是所述样品在所述第二光波长处的总吸光度。
2.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,进一步包括根据具有已知浓度的所述抗体的溶液分别在所述第一光波长或所述第二光波长处的吸光度,计算所述抗体在所述第一光波长处或在所述第二光波长处的消光系数。
3.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,进一步包括根据具有已知浓度的所述药物的溶液分别在所述第一光波长或所述第二光波长处的吸光度,计算所述药物在所述第一光波长处或在所述第二光波长处的消光系数。
4.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中所述尺寸排阻色谱包括超高效液相色谱、反相超高效液相色谱或高效液相色谱。
5.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中所述样品包括1μg/μL至500μg/μL的抗体-药物缀合物。
6.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中所述抗体是抗-肿瘤抗体。
7.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中所述药物是促凋亡剂、细胞生长抑制剂或细胞毒性剂。
8.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中对吸光度响应的峰进行积分包括检测吸光度响应的每个峰的峰肩。
9.根据权利要求1所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的样品中药物与抗体之比的方法,其中所述时间间隔是3分钟至9分钟。
10.一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其包括:
通过以下方法测量包含所述抗体-药物缀合物的第一样品的总吸光度:
将包含所述抗体-药物缀合物的第一样品施加于尺寸排阻色谱基质;
检测所述第一样品在第一光波长λ1处的吸光度响应,其中所述第一光波长是所述抗体的预定吸光度最大值;
检测所述第一样品在第二光波长λ2处的吸光度响应,其中所述第二光波长是所述药物的预定吸光度最大值;
确定所述第一样品在所述第一光波长处的总吸光度和所述第一样品在所述第二光波长处的总吸光度,各个总吸光度通过经由对高分子量峰、低分子量峰和主峰的面积求和对在一定洗脱时间间隔内吸光度响应的峰进行积分来计算;
通过以下方法测量包含所述抗体的第二样品的总吸光度:
将包含所述抗体的第二样品施加于尺寸排阻色谱基质;
检测包含所述抗体的第二样品在所述第一光波长λ1处的吸光度响应;
检测包含所述抗体的第二样品在所述第二光波长λ2处的吸光度响应;以及
确定包含所述抗体的第二样品在所述第一光波长处的总吸光度和所述第二样品在所述第二光波长处的总吸光度,各个总吸光度通过经由对高分子量峰、低分子量峰和主峰的面积求和对在一定洗脱时间间隔内吸光度响应的峰进行积分来计算;以及
使用以下公式2计算DAR:
其中是所述抗体在所述第一光波长处的消光系数;/>是所述药物在所述第一光波长处的消光系数;/>是所述药物在所述第二光波长处的消光系数;/>是包含所述抗体的第二样品在所述第一光波长处的总吸光度;/>是包含所述抗体的第二样品在所述第二光波长处的总吸光度;/>是包含所述抗体-药物缀合物的第一样品在所述第一光波长处的总吸光度;/>是包含所述抗体-药物缀合物的第一样品在所述第二光波长处的总吸光度。
11.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,进一步包括根据具有已知浓度的所述抗体的溶液分别在所述第一光波长或所述第二光波长处的吸光度,计算所述抗体在所述第一光波长处或在所述第二光波长处的消光系数。
12.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,进一步包括根据具有已知浓度的所述药物的溶液分别在所述第一光波长或所述第二光波长处的吸光度,计算所述药物在所述第一光波长处或在所述第二光波长处的消光系数。
13.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中所述尺寸排阻色谱包括超高效液相色谱、反相超高效液相色谱或高效液相色谱。
14.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中所述第一样品包括1μg/μL至500μg/μL的抗体-药物缀合物。
15.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中所述抗体是抗-肿瘤抗体。
16.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中所述药物是促凋亡剂、细胞生长抑制剂或细胞毒性剂。
17.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中对吸光度响应的峰进行积分包括检测吸光度响应的每个峰的峰肩。
18.根据权利要求10所述的一种使用尺寸排阻色谱确定包含抗体-药物缀合物的第一样品中药物与抗体之比的方法,其中所述时间间隔是3分钟至9分钟。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311006893.6A CN117030649A (zh) | 2017-09-08 | 2018-09-05 | 评估抗体-药物缀合物的方法 |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201762556153P | 2017-09-08 | 2017-09-08 | |
US62/556153 | 2017-09-08 | ||
PCT/US2018/049599 WO2019050981A1 (en) | 2017-09-08 | 2018-09-05 | METHODS OF ASSESSING CONJUGATES ANTIBODY-MEDICINE |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311006893.6A Division CN117030649A (zh) | 2017-09-08 | 2018-09-05 | 评估抗体-药物缀合物的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111148985A CN111148985A (zh) | 2020-05-12 |
CN111148985B true CN111148985B (zh) | 2023-08-22 |
Family
ID=63708452
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311006893.6A Pending CN117030649A (zh) | 2017-09-08 | 2018-09-05 | 评估抗体-药物缀合物的方法 |
CN201880058051.6A Active CN111148985B (zh) | 2017-09-08 | 2018-09-05 | 评估抗体-药物缀合物的方法 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311006893.6A Pending CN117030649A (zh) | 2017-09-08 | 2018-09-05 | 评估抗体-药物缀合物的方法 |
Country Status (15)
Country | Link |
---|---|
US (2) | US10976241B2 (zh) |
EP (2) | EP4317980A3 (zh) |
JP (2) | JP2020533565A (zh) |
KR (3) | KR102605352B1 (zh) |
CN (2) | CN117030649A (zh) |
AR (1) | AR112996A1 (zh) |
AU (1) | AU2018328212A1 (zh) |
BR (1) | BR112020000978A2 (zh) |
CA (1) | CA3069455A1 (zh) |
EA (1) | EA202090627A1 (zh) |
IL (3) | IL300065B2 (zh) |
MX (1) | MX2020002395A (zh) |
SG (1) | SG11202000242PA (zh) |
TW (1) | TW201920933A (zh) |
WO (1) | WO2019050981A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7808641B2 (en) * | 2007-04-13 | 2010-10-05 | C Technologies Inc. | Interactive variable pathlength device |
CN111812220A (zh) * | 2020-05-29 | 2020-10-23 | 南京品生医学检验实验室有限公司 | 一种检测血浆中抗肿瘤药物浓度的方法 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7808641B2 (en) * | 2007-04-13 | 2010-10-05 | C Technologies Inc. | Interactive variable pathlength device |
EP2277044B1 (en) * | 2008-05-13 | 2015-06-17 | Genentech, Inc. | Analysis of antibody drug conjugates by bead-based affinity capture and mass spectrometry |
WO2014208987A1 (ko) * | 2013-06-24 | 2014-12-31 | 한화케미칼 주식회사 | 안정성이 개선된 항체-약물 결합체 및 이의 용도 |
US10261058B2 (en) * | 2014-01-14 | 2019-04-16 | Bio-Rad Laboratories, Inc. | Method and system for liquid chromatography data analysis |
-
2018
- 2018-09-05 AR ARP180102519A patent/AR112996A1/es unknown
- 2018-09-05 MX MX2020002395A patent/MX2020002395A/es unknown
- 2018-09-05 EP EP23217146.2A patent/EP4317980A3/en active Pending
- 2018-09-05 BR BR112020000978-4A patent/BR112020000978A2/pt not_active IP Right Cessation
- 2018-09-05 CN CN202311006893.6A patent/CN117030649A/zh active Pending
- 2018-09-05 CA CA3069455A patent/CA3069455A1/en active Pending
- 2018-09-05 WO PCT/US2018/049599 patent/WO2019050981A1/en unknown
- 2018-09-05 JP JP2020509083A patent/JP2020533565A/ja active Pending
- 2018-09-05 CN CN201880058051.6A patent/CN111148985B/zh active Active
- 2018-09-05 EA EA202090627A patent/EA202090627A1/ru unknown
- 2018-09-05 EP EP18779493.8A patent/EP3679344B1/en active Active
- 2018-09-05 IL IL300065A patent/IL300065B2/en unknown
- 2018-09-05 KR KR1020237024975A patent/KR102605352B1/ko active IP Right Grant
- 2018-09-05 US US16/121,909 patent/US10976241B2/en active Active
- 2018-09-05 KR KR1020237039624A patent/KR20230161542A/ko not_active Application Discontinuation
- 2018-09-05 TW TW107131123A patent/TW201920933A/zh unknown
- 2018-09-05 KR KR1020207006712A patent/KR102559940B1/ko active Application Filing
- 2018-09-05 IL IL307618A patent/IL307618A/en unknown
- 2018-09-05 SG SG11202000242PA patent/SG11202000242PA/en unknown
- 2018-09-05 AU AU2018328212A patent/AU2018328212A1/en not_active Abandoned
-
2020
- 2020-01-14 IL IL272028A patent/IL272028B2/en unknown
-
2021
- 2021-02-19 US US17/179,734 patent/US20210172866A1/en active Pending
-
2024
- 2024-02-19 JP JP2024023000A patent/JP2024071381A/ja active Pending
Non-Patent Citations (1)
Title |
---|
Chromatography-based methods for determining molar extinction coefficients of cytotoxic payload drugs and drug antibody ratios of antibody drug conjugates;Wang Chunlei et al.;《Journal of Chromatography A》;20160526;133-139 * |
Also Published As
Publication number | Publication date |
---|---|
EP3679344B1 (en) | 2023-12-27 |
AU2018328212A1 (en) | 2020-02-06 |
JP2020533565A (ja) | 2020-11-19 |
US20210172866A1 (en) | 2021-06-10 |
CA3069455A1 (en) | 2019-03-14 |
IL272028A (en) | 2020-03-31 |
KR20230161542A (ko) | 2023-11-27 |
AR112996A1 (es) | 2020-01-15 |
KR102559940B1 (ko) | 2023-07-27 |
IL300065B2 (en) | 2024-03-01 |
IL300065B1 (en) | 2023-11-01 |
EP3679344A1 (en) | 2020-07-15 |
SG11202000242PA (en) | 2020-02-27 |
EP4317980A3 (en) | 2024-04-17 |
US20190079005A1 (en) | 2019-03-14 |
IL307618A (en) | 2023-12-01 |
EP4317980A2 (en) | 2024-02-07 |
WO2019050981A1 (en) | 2019-03-14 |
EA202090627A1 (ru) | 2020-06-10 |
MX2020002395A (es) | 2020-07-13 |
TW201920933A (zh) | 2019-06-01 |
KR102605352B1 (ko) | 2023-11-24 |
IL300065A (en) | 2023-03-01 |
KR20230113417A (ko) | 2023-07-28 |
CN111148985A (zh) | 2020-05-12 |
KR20200051616A (ko) | 2020-05-13 |
JP2024071381A (ja) | 2024-05-24 |
IL272028B2 (en) | 2023-06-01 |
BR112020000978A2 (pt) | 2020-07-14 |
US10976241B2 (en) | 2021-04-13 |
CN117030649A (zh) | 2023-11-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP2024071381A (ja) | 抗体-薬物複合体の評価方法 | |
Wankhede et al. | Development and validation of UV-spectrophotometric methods for simultaneous estimation of cetirizine hydrochloride and phenylephrine hydrochloride in tablets | |
Bhongade et al. | A validated method for the quantitation of ciprofloxacin hydrochloride using diffuse reflectance infrared fourier transform spectroscopy | |
EP2344265B1 (en) | Evaluating heparin preparations for pharmaceutical use | |
Broto et al. | Bioanalytical methods for cytostatic therapeutic drug monitoring and occupational exposure assessment | |
Young et al. | Evaluation of the physicochemical and functional stability of diluted REMSIMA® upon extended storage—a study compliant with NHS (UK) guidance | |
SAVALE | UV spectrophotometric method development and validation for quantitative estimation of rutin | |
Reddy et al. | Determination of edoxaban in bulk and in tablet dosage form by stability indicating high-performance liquid chromatography | |
US10444144B2 (en) | Identification of consumed drugs and food by unique near infrared tag libraries | |
Bhaskar et al. | Q-analysis and simultaneous equation method for estimation of domperidone and naproxen by UV spectrophotometry in bulk and tablet dosage form | |
Mojsiewicz-Pieńkowska | Size exclusion chromatography with evaporative light scattering detection as a method for speciation analysis of polydimethylsiloxanes. II. Validation of the method for analysis of pharmaceutical formulations | |
Syahputra et al. | Determination of simultaneous irbesartan and hydrochlorothiazide by ultraviolet spectrophotometry with dual wavelength method | |
Vetrichelvan et al. | New Spectrophotometric Methods for the Determination of Racecadotril in Bulk Drug and Capsules. | |
Rajanit et al. | Absorption Correction Method for Simultaneous Estimation of Nifedipine and Metoprolol Succinate in Their Synthetic Mixture Using from Spectrophotometry | |
Vijaya et al. | Validation of UV Spectrophotometric and HPLC Methods for quantitative determination of Imatinib in Bulk and Pharmaceutical Dosage Form | |
Bharatea et al. | Development of validated spectrophotometric method for simultaneous estimation of acetylsalicylic acid and caffeine in pure and tablet dosage form | |
JP6734489B1 (ja) | 生体液の検査方法 | |
Rele et al. | Spectrophotometric Estimation of Carbocisteine in Bulk and Pharmaceutical Dosage Form by First Order Derivative and Area Under Curve Methods | |
Das et al. | Analytical Method for Development and Validation of Flupirtine Maleate by Reverse-phase High-performance Liquid Chromatography | |
Gobi | Development Newer Analytical Techniques for the Estimation of Benfotiamine and its Combination with Pyridoxamine Dihydrochloride and Resveratrol | |
Chinnapurkar et al. | DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR DETERMINATION OF RIFABUTINE IN API AND CAPSULE DOSAGE FORM | |
Khan et al. | Evaluation of the physicochemical and functional | |
Bhandary et al. | NON-COMPARTMENTAL PHARMACOKINETICS MODELING OF AMLODIPINE IN RATS | |
VENUMADHAV et al. | NEW SPECTROPHOTOMETRIC METHODS FOR THE DETERMINATION OF LORNOXICAM IN PHARMACEUTICAL DOSAGE FORMS. E. VENUMADHAV1, T. NEEHA2, P. BHARGAVI2, AMREEN NISHAT2, A. SWETHA2 AND G. DEVALA RAO2 | |
Vigbedor | Design of HPLC method and stability studies of cyanocobalamin injection |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |