Abstract

Estradiol-17β-glucuronide (E₂17G) is an estrogen metabolite that has cholestatic properties. In humans, circulating E₂17G is transported into hepatocytes by organic anion transporting polypeptides (OATPs) and is excreted into bile by multidrug-resistance associated protein 2 (MRP2). E₂17G is also a substrate of the basolateral efflux transporters MRP3 and MRP4, which translocate E₂17G from hepatocytes to blood. However, the contribution of basolateral efflux to hepatocyte disposition of E₂17G has not been evaluated previously. To address this question, E₂17G disposition was studied in sandwich-cultured human hepatocytes and mechanistic modeling was applied to calculate clearance values (mean ± S.D.) for uptake, intrinsic biliary excretion (CL_int,bile) and intrinsic basolateral efflux (CL_int,BL). The biliary excretion index of E₂17G was 45% ± 6%. The CL_int,BL of E₂17G [0.18 ± 0.03 (ml/min)/g liver) was 1.6-fold higher than CL_int,bile [0.11 ± 0.06 (ml/min)/g liver]. Simulations were performed to study the effects of increased CL_int,BL and a concomitant decrease in CL_int,bile on hepatic E₂17G exposure. Results demonstrated that increased CL_int,BL can effectively reduce hepatocellular and biliary exposure to this potent cholestatic agent. Simulations also revealed that basolateral efflux can compensate for impaired biliary excretion and, vice versa, to avoid accumulation of E₂17G in hepatocytes. However, when both clearance processes are impaired by 90%, hepatocyte E₂17G exposure increases up to 10-fold. These data highlight the contribution of basolateral efflux transport, in addition to MRP2-mediated biliary excretion, to E₂17G disposition in human hepatocytes. This elimination route could be important, especially in cases where basolateral efflux is induced, such as cholestasis.