A sensitive LC–MS/MS assay to quantitatefree payload Aur0101 from ADC PYX-201 inrat and monkey plasma: supplementary materials

Aim: Aur0101 is a cytotoxic and small-molecule microtubule depolymerizing agent, and is the payload conjugated to antibody–drug conjugate PYX-201. Developing and validating a sensitive bioanalytical method to quantitate Aur0101 was novel and crucial in preclinical PYX-201 studies. Materials & methods: Reference standard Aur0101 and its stable isotope labelled internal standard Aur0101-d8 were used in this LC–MS/MS method. Results: This sensitive assay was validated at a lower limit of quantitation of 15 pg/ml and successfully applied to support preclinical rat and monkey toxicology studies. Preclinical plasma toxicokinetic parameters were presented. Conclusion: A sensitive and robust LC–MS/MS assay was validated for Aur0101 in rat and monkey plasma.研究目的：Aur0101是一种细胞毒性小分子微管解聚剂，同时作为抗体药物偶联物（antibody–drug conjugate）PYX-201的偶联有效载荷。在PYX-201的临床前研究中，建立并验证一种可定量检测Aur0101的高灵敏度生物分析方法，具有重要的创新性与关键意义。 材料与方法：本研究采用Aur0101对照品及其稳定同位素标记内标Aur0101-d8，构建液相色谱-串联质谱（LC–MS/MS）检测方法。 结果：该高灵敏度检测方法的定量下限为15 pg/ml，经方法学验证后成功应用于大鼠与猴的临床前毒理学研究，并报道了相关临床前血浆毒代动力学参数。 结论：本研究成功建立并验证了一种可用于大鼠与猴血浆中Aur0101定量检测的高灵敏度、稳健型LC–MS/MS检测方法。