Hepatic basolateral efflux contributes significantly to rosuvastatin disposition I: characterization of basolateral versus biliary clearance using a novel protocol in sandwich-cultured hepatocytes

J Pharmacol Exp Ther. 2013 Dec;347(3):727-36. doi: 10.1124/jpet.113.207472. Epub 2013 Sep 10.


Transporters responsible for hepatic uptake and biliary clearance (CLBile) of rosuvastatin (RSV) have been well characterized. However, the contribution of basolateral efflux clearance (CLBL) to hepatic and systemic exposure of RSV is unknown. Additionally, the appropriate design of in vitro hepatocyte efflux experiments to estimate CLBile versus CLBL remains to be established. A novel uptake and efflux protocol was developed in sandwich-cultured hepatocytes (SCH) to achieve desired tight junction modulation while maintaining cell viability. Subsequently, studies were conducted to determine the role of CLBL in the hepatic disposition of RSV using SCH from wild-type (WT) and multidrug resistance-associated protein 2 (Mrp2)-deficient (TR(-)) rats in the absence and presence of the P-glycoprotein and breast cancer resistance protein (Bcrp) inhibitor elacridar (GF120918). RSV CLBile was nearly ablated by GF120918 in TR(-) SCH, confirming that Mrp2 and Bcrp are responsible for the majority of RSV CLBile. Pharmacokinetic modeling revealed that CLBL and CLBile represent alternative elimination routes with quantitatively similar contributions to the overall hepatocellular excretion of RSV in rat SCH under baseline conditions (WT SCH in the absence of GF120918) and also in human SCH. Membrane vesicle experiments revealed that RSV is a substrate of MRP4 (Km = 21 ± 7 µM, Vmax = 1140 ± 210 pmol/min per milligram of protein). Alterations in MRP4-mediated RSV CLBL due to drug-drug interactions, genetic polymorphisms, or disease states may lead to changes in hepatic and systemic exposure of RSV, with implications for the safety and efficacy of this commonly used medication.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acridines / pharmacology
  • Adenosine Triphosphate / physiology
  • Algorithms
  • Animals
  • Bile / metabolism*
  • Carrier Proteins / metabolism
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Fluorobenzenes / pharmacokinetics*
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism*
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacokinetics*
  • L-Lactate Dehydrogenase / metabolism
  • Male
  • Models, Biological
  • Multidrug Resistance-Associated Proteins / metabolism
  • Plasmids
  • Pyrimidines / pharmacokinetics*
  • Rats
  • Rats, Wistar
  • Rosuvastatin Calcium
  • Sulfonamides / pharmacokinetics*
  • Tetrahydroisoquinolines / pharmacology
  • Tight Junctions / metabolism


  • Abcc4 protein, rat
  • Acridines
  • Carrier Proteins
  • Fluorobenzenes
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • Multidrug Resistance-Associated Proteins
  • Pyrimidines
  • Sulfonamides
  • Tetrahydroisoquinolines
  • multidrug resistance-associated protein 3
  • Rosuvastatin Calcium
  • Adenosine Triphosphate
  • L-Lactate Dehydrogenase
  • Elacridar