Evaluation of Normothermic Machine Perfusion of Porcine Livers as a Novel Preclinical Model to Predict Biliary Clearance and Transporter-Mediated Drug-Drug Interactions Using Statins.

L J Stevens,P P Chothe,W H J Vaes,J M Donkers,C A J Knibbe,E Van De Steeg,A Z X Zhu,S K Chowdhury,I P J Alwayn

doi:10.1124/dmd.121.000521

Abstract

There is a lack of translational preclinical models that can predict hepatic handling of drugs. In this study, we aimed to evaluate the applicability of normothermic machine perfusion (NMP) of porcine livers as a novel ex vivo model to predict hepatic clearance, biliary excretion, and plasma exposure of drugs. For this evaluation, we dosed atorvastatin, pitavastatin, and rosuvastatin as model drugs to porcine livers and studied the effect of common drug-drug interactions (DDIs) on these processes. After 120 minutes of perfusion, 0.104 mg atorvastatin (n = 3), 0.140 mg pitavastatin (n = 5), or 1.4 mg rosuvastatin (n = 4) was administered to the portal vein, which was followed 120 minutes later by a second bolus of the statin coadministered with OATP perpetrator drug rifampicin (67.7 mg). After the first dose, all statins were rapidly cleared from the circulation (hepatic extraction ratio > 0.7) and excreted into the bile. Presence of human-specific atorvastatin metabolites confirmed the metabolic capacity of porcine livers. The predicted biliary clearance of rosuvastatin was found to be closer to the observed biliary clearance. A rank order of the DDI between the various systems upon coadministration with rifampicin could be observed: atorvastatin (AUC ratio 7.2) > rosuvastatin (AUC ratio 3.1) > pitavastatin (AUC ratio 2.6), which is in good agreement with the clinical DDI data. The results from this study demonstrated the applicability of using NMP of porcine livers as a novel preclinical model to study OATP-mediated DDI and its effect on hepatic clearance, biliary excretion, and plasma profile of drugs. SIGNIFICANCE STATEMENT: This study evaluated the use of normothermic machine perfusion (NMP) of porcine livers as a novel preclinical model to study hepatic clearance, biliary excretion, plasma (metabolite) profile of statins, and OATP-mediated DDI. Results showed that NMP of porcine livers is a reliable model to study OATP-mediated DDI. Overall, the rank order of DDI severity indicated in these experiments is in good agreement with clinical data, indicating the potential importance of this new ex vivo model in early drug discovery.

Highlights

The liver is a complex organ involved in the uptake, metabolism, and biliary excretion of xenobiotics and endogenous compounds
We describe for the first time the use of normothermic machine perfusion (NMP) of porcine livers to predict human hepatobiliary disposition and drug-drug interaction (DDI) of drugs as demonstrated by using known organic anion transporting peptides (OATP) substrate drugs such as atorvastatin, pitavastatin, and rosuvastatin with known OATP inhibitor rifampicin as a perpetrator drug
Our results showed a relation between plasma rifampicin area under the concentration-time curve (AUC) and AUC ratio (AUCR) for atorvastatin, whereas minor variation was observed for the pitavastatin and Percent Biliary Clearance of Metabolites

Summary

Introduction

The liver is a complex organ involved in the uptake, metabolism, and biliary excretion of xenobiotics and endogenous compounds. Concomitant administration of drugs that are substrates for the same transporters and/or metabolizing enzymes can result in a drug-drug interaction (DDI) affecting plasma as well as biliary levels of one of the drugs As a result, this can change the drug concentration at ABBREVIATIONS : ALT, alanine aminotransferase; AST, aspartate aminotransaminase; AUC, area under the concentration-time curve; AUCR, AUC ratio; BCRP, breast cancer resistance protein; BSEP, bile salt export pump; DDI, drug-drug interaction; GLUT1, glucose transporter 1; ICG, indocyanine green; IPRL, isolated perfused rat liver; MCT1, monocarboxylate transporter 1; MDR1, multidrug resistance protein 1; MRP1, multidrug resistance–associated protein 1; MRP2, multidrug resistance–associated protein 2; MRP3, multidrug resistance–associated protein 3; NMP, normothermic machine perfusion; NTCP, Na1-taurocholate cotransporting polypeptide; OATP, organic anion transporting polypeptide; OATP1B1, organic anion transporting polypeptide 1B1; OATP1B3, organic anion transporting polypeptide 1B3; OATP2B1, organic anion transporting polypeptide 2B1; PK, pharmacokinetics

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