Novel multiple assessment of hepatocellular drug disposition in a single packaged procedure

Abstract

Better prediction of drug disposition prior to the clinical trial is critical for the efficient development of new drugs. The purpose of this study is to develop a novel multiple assessment methodology of hepatocellular drug disposition from drug uptake to efflux including biliary and basolateral excretion, in a single packaged procedure. We started a sandwich culture using rat primary hepatocytes. After five days culture, the hepatocytes were incubated with a dosing solution including CDF or Rhodamine 123. Three distinct sequences were then performed in parallel: disrupting and maintaining the tight junctions comprising a bile canalicular network at 37 °C, and maintaining the network at 4 °C. Supernatant fractions were collected from each sequence, and followed by the cell lysate collection. The disposition rates of basolateral efflux by diffusion, by transporter-mediation, biliary excretion, and residual cellular fraction of CDF and Rhodamine 123 were 38.2% and 11.0%, 26.6% and 12.1%, 18.6% and 4.9%, and, 16.7% and 72.0%, respectively. CDF was likely to excrete extracellularly whereas Rhodamine 123 tended to remain intracellularly. CDF showed a relatively higher biliary excretion rate than Rhodamine 123. This novel protocol may contribute to improve the predictability of pharmacokinetics eventually in human, and streamline new drug development. Chemical compounds5(6)-Carboxy-2′,7′-dichlorofluoroscein (PubChem CID: 132525); Rhodamine 123 (PubChem CID: 65217).