title: 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.
  5(6)-Carboxy-2',7'-dichlorofluoroscein (PubChem CID: 132525); Rhodamine 123 (PubChem CID: 65217).
authors.affiliation: Center for Technology Innovation - Healthcare, Hitachi, Ltd., 2520 Hatoyama, Saitama 350-0395, Japan. Electronic address: ryosuke.takahashi.jr@hitachi.com.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Takahashi
authors.affiliation: Center for Technology Innovation - Healthcare, Hitachi, Ltd., 2520 Hatoyama, Saitama 350-0395, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Ichikawa
authors.affiliation: Science Systems Design Div., Hitachi High-Technologies Corporation, 882, Ichige, Hitachinaka, Ibaraki 312-8504, Japan. Electronic address: katsuhiro.kanda.pv@hitachi-hightech.com.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Kanda
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005452:Fluoresceins / Q000493:pharmacokinetics*; D022781:Hepatocytes / Q000378:metabolism*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D020112:Rhodamine 123 / Q000493:pharmacokinetics*
chemical_list: D005452:Fluoresceins; C053897:5(6)-carboxy-2',7'-dichlorofluorescein; D020112:Rhodamine 123
keywords: Basolateral efflux; Biliary excretion; Drug disposition; Hepatocyte; Sandwich culture; Uptake