Successful Prediction of In Vivo Hepatobiliary Clearances and Hepatic Concentrations of Rosuvastatin Using Sandwich-Cultured Rat Hepatocytes, Transporter-Expressing Cell Lines, and Quantitative Proteomics

Abstract

We determined whether in vivo transporter-mediated hepatobiliary clearance (CL) and hepatic concentrations of rosuvastatin (RSV) in the rat could be predicted by transport activity in sandwich-cultured rat hepatocytes (SCRHs) and/or transporter-expressing cell lines scaled by differences in transporter protein expression between SCRHs, cell lines, and rat liver. The predicted hepatobiliary CLs and hepatic concentrations of RSV were compared with our previously published positron emission tomography imaging data. Sinusoidal uptake CL ([Formula: see text] [Formula: see text] [Formula: see text] [Formula: see text] [Formula: see text] [Formula: see text] for Oatp protein expression. This is the first demonstration of the successful prediction of in vivo hepatobiliary CLs and hepatic concentrations of RSV using transporter-expressing cells and SCRHs.

Positron Emission Tomography Imaging of [11C]Rosuvastatin Hepatic Concentrations and Hepatobiliary Transport in Humans in the Absence and Presence of Cyclosporin A

“…However, except for hepatocytes, primary cells from other tissues (e.g., blood–brain barrier and kidney epithelial proximal tubule cells) are either not routinely available or validated. Moreover, even where available (e.g., hepatocytes), they may not be predictive of the in vivo CL of the drug of interest . The reason for this lack of prediction is not well known but could be that the abundance and/or activity of the transporters in the cells may not represent that in vivo .…”

“…To address the shortcomings of the above approaches, our laboratory has hypothesized that drug concentration in tissues can be predicted from in vitro CL studies in cells that individually express the transporters of interest. Then, these CL values can be scaled to that in vivo ( in vitro – in vivo extrapolation (IVIVE)) using the proteomics‐informed bottom‐up approach . However, such predictions must be verified to determine their accuracy.…”

“…Therefore, we propose here that such verification be conducted for selected “probe” substrates that interrogate the major transporters present in the tissue of interest. We have shown the proof‐of‐principle of this approach by successfully predicting the hepatic concentrations of rosuvastatin (RSV) in the rat (obtained by PET imaging) by predicting the sinusoidal influx (organic‐anion‐transporting polypeptide (OATP)‐mediated) and sinusoidal efflux CL and biliary efflux CL of RSV in cells expressing the relevant transporters . As a first step toward replicating this proof‐of‐principle in humans, we conducted for the first time [ 11 C]RSV PET imaging in humans.…”

“…Then, these CL values can be scaled to that in vivo (in vitro-in vivo extrapolation (IVIVE)) using the proteomics-informed bottom-up approach. 9 However, such predictions must be verified to determine their accuracy. This cannot be done for every drug under development.…”

Toward a Consensus on Applying Quantitative Liquid Chromatography‐Tandem Mass Spectrometry Proteomics in Translational Pharmacology Research: A White Paper

“…In this approach, if both datasets are generated in the same laboratory and relative protein extraction and digestion efficiencies for cell and tissue samples are similar, a relative expression factor can be established among systems for IVIVE ( Table ). This approach has been successfully used for IVIVE of transporter‐based clearance of drugs …”