Abstract

Despite the fact that much progress has been made recently in the development of targeted covalent inhibitors (TCIs), their pharmacokinetics (PK) have not been well characterized in the light of extrahepatic clearance (CL_extH) by glutathione (GSH)/glutathione S-transferase (GST)-dependent conjugation attributable to the unique electrophilic structure (e.g., acrylamide moiety) of TCI compounds. In the present study, CL_extH values were examined in rat, dog, and monkey to predict the contribution of CL_extH to the PK of the TCIs afatinib, ibrutinib, and neratinib in humans. Afatinib and neratinib both underwent extensive conjugation with GSH in buffer and cytosol fractions of liver and kidney, whereas ibrutinib showed much lower reactivity/susceptibility to GSH/GST-dependent conjugation. The CL_extH in each species was calculated from the difference between observed total body clearance and predicted hepatic clearance (CL_H) in cryopreserved hepatocytes suspended in 100% serum of the corresponding species. The power-based simple allometry relating the CL_extH for the unbound compound to animal body weight was applicable across species for afatinib and neratinib (R² ≥ 0.9) but not for ibrutinib (R² = 0.04). The predicted AUC after oral administration of afatinib and neratinib agreed reasonably closely with reported values in phase I dose-escalation studies. Comparisons of CL_extH and CL_H predicted that CL_extH largely determined the PK of afatinib (>90% as a proportion of total body clearance) and neratinib (∼34%) in humans. The present method can serve as one of the tools for the optimization of PK in humans at the discovery stage for the development of TCI candidates.