Improved Predictability of Hepatic Clearance with Optimal pH for Acyl-Glucuronidation in Liver Microsomes

The purpose of this study was to investigate the optimal pH for acyl-glucuronidation formation with carboxylic acid-containing compounds in human and rat liver microsomes to improve the predictability of their hepatic clearance. The optimal pH for acyl-glucuronidation of all 17 compounds was around pH 6.0 in human and rat liver microsomes. Correlation analysis was done with the predicted in vitro intrinsic clearance (CL_{int,in vitro}) and in vivo intrinsic clearance (CL_{int,in vivo}) calculated from available reported data of total clearance (CL_tot) of 11 compounds in humans. For 8 of the 11 compounds, under the pH 6.0 condition, the CL_{int,in vitro} were within 1/3 to 3-fold error of the observed CL_{int,in vivo} whereas, the error was within 1/3 to 3-fold of the observed CL_{int,in vivo} for only 3 of the 11 under the pH 7.4 condition. The intracellular pH in human and rat hepatocytes decreased in the presence of a carboxylic acid-containing compound. These findings suggest that acyl-glucuronidation in liver microsomes at pH 6.0 is closer to physiological conditions in the presence of carboxylic acid compounds, and thus, use of this pH condition is important for physiological interpretation and predictability of intrinsic clearance.