ABSTRACT
The stability and exposure of toxin-related catabolites in system circulation contributes to the evaluation of the stability, targeted delivery and off-target toxicity for antibody-drug conjugates (ADC) at different stages during drug development. In this study, simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for determination catabolites of Mertansine (DM1), MCC-DM1 and Lys-MCC-DM1 in cynomolgus serum have been developed. The serum samples are processed by protein precipitation. The LC-MS/MS methods are applied on a Phenomenex C8 column (50 × 2.0 mm, 5 µm) with gradient elution with water-formic acid 0.1% (A) and acetonitrile-formic acid 0.1% (B) at a flow rate of 0.5 mL/min. The analytical run time is only 4.0 min and the calibration ranges of the standard curve are 0.500-200 ng/mL for DM1, 1.00-500 ng/mL for MCC-DM1 and 2.00-1000 ng/mL for Lys-MCC-DM1. Intra- and inter-day precision of low, middle and high quality controls was <15%, and accuracy was 99.2-110.9%. The methods were successfully applied to evaluate three catabolites of novel ADCs with N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate linker in vitro and in vivo studies.
