title: A Modified Procedure for Estimating the Impact of the Uptake on the Overall Biliary Clearance in Sandwich Culture of Rat Hepatocytes.
abstract: Sandwich culture of hepatocytes is commonly applied for the prediction of in vivo biliary clearance (CLbil). In this paper, we present a modified procedure for the determination of in vitro CLbil in sandwich culture of rat hepatocytes, which allows the estimation of the impact of uptake processes on the overall CLbil. The main point of this modification is the separation of uptake and efflux processes. Ten drugs from four biopharmaceutics drug disposition classification system classes were chosen in order to demonstrate the advantages of this method: 1) the uptake is performed identically before the canaliculi are opened, thus the efflux starts at the same intracellular concentration of the drugs and the effect of Ca2+/Mg2+ depletion on the uptake is excluded; 2) exact intracellular concentrations can be measured at the start and at the end of the efflux; 3) the biliary clearance can be determined irrespective of the uptake; 4) the canalicular and the sinusoidal transport can be measured simultaneously; 5) drug-drug interactions concerning uptake and efflux transporters can be estimated independently. Depending on the degree of uptake, CLbil,app (calculated using the concentration of drugs in the medium) was significantly higher (sulfasalazine, fluvastatin, rosuvastatin, atorvastatin) or lower (pravastatin, procainamide) than CLbil,int (calculated using the intracellular concentration of drugs). When the uptake had no impact on the CLbil, the apparent and intrinsic CLbil did not differ significantly (lovastatin, rifampicin, quetiapine). Our results confirm that transporters may play a significant role in the uptake of drugs both with high and poor permeability and solubility.
authors.affiliation: Department of Functional Pharmacology, Institute of Cognitive Neuroscience and Psychology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Address: H-1117 Budapest, Magyar tudosok krt 2, Hungary. jemnitz.katalin@ttk.mta.hu.
authors.forename: Katalin
authors.initials: K
authors.lastname: Jemnitz
authors.forename: Monika
authors.initials: M
authors.lastname: Szabo
authors.forename: Attila
authors.initials: A
authors.lastname: Batai-Konczos
authors.forename: Pal
authors.initials: P
authors.lastname: Szabo
authors.forename: Balazs
authors.initials: B
authors.lastname: Magda
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Veres
mesh_terms: D000818:Animals; D001646:Bile / Q000378:metabolism*; D002478:Cells, Cultured; D004347:Drug Interactions; D065669:Hepatobiliary Elimination; D022781:Hepatocytes / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D026901:Membrane Transport Proteins / Q000378:metabolism; D008954:Models, Biological; D010539:Permeability; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D061251:Primary Cell Culture; D017208:Rats, Wistar; D012995:Solubility
chemical_list: D026901:Membrane Transport Proteins; D004364:Pharmaceutical Preparations