Organic anion transporting polypeptide 2-mediated uptake of paclitaxel and 2′-ethylcarbonate-linked paclitaxel in freshly isolated rat hepatocytes

Abstract

AbstractObjectivesThe P-glycoprotein (P-gp) efflux pump plays an important role in paclitaxel detoxification. However, hepatic uptake of paclitaxel mediated by a solute-linked carrier transporter family is still poorly understood in animals and humans. Freshly isolated hepatocyte suspensions are a well established in-vitro model for studying drug transport and xenobiotic metabolism. Therefore, the hepatic uptake of paclitaxel and its P-gp-insensitive prodrug, 2′-ethylcarbonate-linked paclitaxel (TAX-2′-Et), has been characterized using freshly isolated and pregnenolone-16-α-carbonitrile (PCN)-treated hepatocytes in rats.MethodsPaclitaxel and TAX-2′-Et were incubated with rat hepatocyte suspensions in the presence or absence of inhibitors.Key findingsPaclitaxel and TAX-2′-Et showed concentration-dependent uptake in rat hepatocytes. The intrinsic transport capacity was two-fold higher for paclitaxel uptake than for TAX-2′-Et uptake. Rifampicin (a potent inhibitor of organic anion transporting polypeptide (Oatp) 2), but not indometacin (a representative inhibitor of organic anion transporter (Oat) 2 and Oatp1) treatment, significantly inhibited the uptake of paclitaxel and TAX-2′-Et. We characterized the rifampicin-sensitive uptake of paclitaxel and TAX-2′-Et using rat hepatocytes treated with PCN, which dramatically enhances hepatic Oatp2 protein levels. PCN-treated hepatocytes displayed a 1.6-fold greater uptake of paclitaxel and TAX-2′-Et than the vehicle-treated hepatocytes. The uptake of the two compounds was significantly reduced by rifampicin but not by indometacin treatment. These findings demonstrated that the rat Oatp2, but not Oatp1 orOat2, was a candidate transporter for the hepatic uptakeofpaclitaxel and TAX-2′-Et.ConclusionsThe findings have provided an important step towards identifying a key transporter in hepatic detoxification of paclitaxel and TAX-2′-Et in small animals.