In vivo hepatic clearance of lipophilic drugs predicted by in vitro uptake data into cryopreserved hepatocytes suspended in sera of rats, guinea pigs, monkeys and humans.

Evaluation of uptake of lipophilic acid compounds into hepatocytes was an unresolved drug development issue because of their adsorption to cells and materials and low analytical sensitivity and accuracy in assessment of protein bindings. Uptake assays of compounds using hepatocytes suspended in serum were expected to solve these problems for prediction of in vivo hepatic clearance. Here, for compounds with high protein binding (>99%), diflunisal, montelukast, cerivastatin, telmisartan, fluvastatin and six new drug candidates, in vivo hepatic clearance predicted based on hepatic depletion and uptake (CLh, uptake, predicted) data using hepatocytes in the absence and presence of sera was investigated. In vitro hepatic uptake results with hepatocytes suspended in serum improved prediction of human hepatic clearance values for highly lipophilic montelukast and telmisartan. In vivo CLh, uptake, predicted values of six new highly lipophilic acid drug candidates (protein binding >99.97%) and diflunisal, montelukast and cerivastatin predicted based on hepatocytes suspended in serum were within threefold differences of their total clearance in vivo in rats, guinea pigs or monkeys, except for montelukast in monkeys (5.8-fold). These results suggest that the human hepatic uptake in hepatocytes suspended in serum is useful for prediction of CLh, uptake, predicted, especially for highly lipophilic/protein binding acid compounds.