A sample preparation and detection strategy for quantifying proteolytically unstable therapeutic peptides for early-ADME tissue distribution studies

摘要

THR-099 is a prototype peptide drug candidate for the treatment of chronic kidney disease (CKD). In this study, the effects of tissue sampling prior to sample preparation and bioanalysis were investigated. An optimized organ processing, sample preparation, and analysis workflow was developed in order to accurately quantitate THR-099 in mouse kidney. A novel heat stabilization device (Denator Stabilizor T1) was used to inactivate proteases, and compared to conventional snap-freezing approaches. When comparing both approaches from kidneys obtained from the same animal, up to 5 times more intact THR-099 was detected in heat stabilized vs. snap-frozen tissue.