Evaluation of the clearance and metabolic profile of low clearance compounds using HR-MS and plated cryopreserved hepatocytes

摘要

The method presented here using plate human hepatocytes and UPLC-HRAMS is a valid means of determining the apparent in vitro intrinsic clearance of low clearance compounds. These values are used to predict in vivo clearance values, which are key in estimating doses for first in human studies. Underprediction and large intersubject variability of clearance is observed with plated hepatocytes, as with hepatocyte suspensions. 2. Statistical analysis of elimination constants indicates that while hepatocyte suspension incubations (acceptable enzyme activity for ≤4h) can accurately measure half-life values of ～1h or less, plated hepatocyte incubations can measure half -life values of ～60h. The minimum predicted clearance achievable translates to roughly 0.5-1 ml/min/kg (assumes a slope error of 0.005 with 7 timepoints and p<0.05).