Development of a Lc–Ms/Ms Method for the Quantification of Toxic Payload Dm1 Cleaved from Bt1718 in a Phase I Study
Catherine Gowland, Philip Berry, Julie Errington, Philip D. Jeffrey, Gavin Bennett, Lisa Godfrey, Marc Pittman, Andrew Niewiarowski, Stefan N. Symeonides, Gareth J. Veal
Abstract
Background: BT1718 is a novel bicyclic peptide anticancer drug targeting membrane type I matrix metalloproteinase to release its toxic payload DM1. A LC–MS/MS method was validated to quantify DM1 generated from BT1718 in a Phase I/IIa clinical trial. Materials & methods: Plasma samples underwent a reduction reaction to artificially cleave BT1718 into DM1 and its bicycle components. An alkylation step was carried out to stabilize the reaction products, and plasma proteins extracted using acetonitrile. LC–MS/MS analysis utilized a C18 column and Agilent 6460 triple quadrupole mass spectrometer (Agilent, Cheshire, UK). Results: The method was fully validated over a linear range of 200–50,000 ng/ml BT1718, with overall precision ≤10% and accuracy 89–102%. Conclusion: A novel method for quantifying DM1 yielded from BT1718 has been validated and is now being utilized clinically.