title: In vivo drug-linker stability of an anti-CD30 dipeptide-linked auristatin immunoconjugate.
abstract: Effective antibody-drug conjugates (ADC) combine high drug-linker stability in circulation and efficient intratumoral release of drug. Conjugation of monomethyl auristatin E (MMAE) to the anti-CD30 monoclonal antibody (mAb), cAC10, produced a selective and potent ADC against CD30(+) anaplastic large cell lymphoma and Hodgkin's disease models. This ADC, cAC10-valine-citrulline-MMAE, uses a protease-sensitive dipeptide linker designed to release MMAE by lysosomal cathepsin B in target cells but maintain a stable linkage and attenuate drug potency in circulation. To evaluate ADC stability in vivo, we developed methods for measuring drug/mAb ratios at progressive times in plasma from ADC-treated mice and nonhuman primates. Anti-idiotype mAb permitted the capture and quantitation of mAb cAC10, whereas antidrug mAb and MMAE-conjugated horseradish peroxidase reporter provided quantitative detection of conjugated drug following its in vitro release by cathepsin B. These data were validated by an alternative ELISA using anti-idiotype and anti-MMAE mAbs for capture and detection, respectively. Both methods differentiated ADC with variable levels of drug loading and were subsequently applied to stability studies in severe combined immunodeficient mice and cynomolgus monkeys. Evaluation of ADC from mouse circulation showed the linker half-life to be approximately 144 hours (6.0 days), significantly greater than that reported for disulfide- or hydrazone-linked ADCs in mice or human trials. In cynomolgus monkey, the apparent linker half-life was approximately 230 hours (9.6 days), suggesting that the drug-linker will be highly stable in humans. These data represent the longest reported drug-linker half-life to date and provide the basis for the pronounced specificity and antitumor activity of cAC10-valine-citrulline-MMAE.
authors.affiliation: Seattle Genetics Inc., 21813 30th Drive Southeast, Bothell, WA 98021, USA.
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Sanderson
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Hering
authors.forename: Stephanie F
authors.initials: SF
authors.lastname: James
authors.forename: Michael M C
authors.initials: MM
authors.lastname: Sun
authors.forename: Svetlana O
authors.initials: SO
authors.lastname: Doronina
authors.forename: Anthony W
authors.initials: AW
authors.lastname: Siadak
authors.forename: Peter D
authors.initials: PD
authors.lastname: Senter
authors.forename: Alan F
authors.initials: AF
authors.lastname: Wahl
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000627:therapeutic use; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004220:Disulfides; D004355:Drug Stability; D005260:Female; D006689:Hodgkin Disease / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism; D006835:Hydrazones; D018796:Immunoconjugates / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use*; D017730:Ki-1 Antigen / Q000276:immunology* / Q000378:metabolism; D017728:Lymphoma, Large-Cell, Anaplastic / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism; D008252:Macaca fascicularis; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D009842:Oligopeptides / Q000737:chemistry / Q000627:therapeutic use*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D004220:Disulfides; D006835:Hydrazones; D018796:Immunoconjugates; D017730:Ki-1 Antigen; D009842:Oligopeptides; C495575:monomethyl auristatin E