Both P-gp and MRP2 mediate transport of Lopinavir, a protease inhibitor

Abstract

Polarized epithelial non-human (canine) cell lines stably transfected with human or murine complementary DNA (cDNA) encoding for various efflux transporters (P-gp/MDR1, MRP1, MRP2, and Bcrp1) were used to study transepithelial transport of Lopinavir (LVR) and compare results with the MDCKII-Wild type cells. These transmembrane proteins cause multidrug resistance by decreasing the total intracellular accumulation of drugs. Lopinavir efflux was directional and was completely inhibited by MK-571, a selective MRP family inhibitor in the MDCKII-MRP2 cell line. Similarly, LVR efflux was also inhibited by P-gp inhibitors P-gp 4008 and GF120918 in the MDCKII-MDR1 cell line. The efflux ratios (Efflux rate/ Influx rate) of LVR in the absence of any efflux inhibitors in the MDCK-Wild type, MDCKII-MDR1, MDCKII-MRP1 and MDCKII-MRP2 cell monolayers were 1.32, 4.91, 1.26 and 2.89 respectively. The MDCKII-MDR1 and MDCKII-MRP2 cells have significantly increased LVR efflux ratio relative to the parental cells due to the apically directed transport by MDR1 and MRP2 respectively. The efflux ratios in MRP2 and MDR1 transfected cell lines were close to unity in the presence of MK-571 and P-gp 4008 respectively; indicating that LVR efflux by MRP2 and P-gp was completely inhibited by their selective inhibitors. MDCKII-MRP1 cells did not exhibit a significant reduction in the LVR efflux relative to the parental cells, indicating that LVR is not a good substrate for MRP1. Transport studies across MDCKII-Bcrp1 cells indicated that LVR is not transported by Bcrp1 and is not a substrate for this efflux protein. In conclusion, this study presents direct evidence that LVR is effluxed by both P-gp and MRP2 which may contribute to its poor oral bioavailability and limited penetration into the CNS. KeywordsMDCKII-MDR1; MDCKII-MRP2; MDCKII-MRP1; MDCKII-Bcrp1; MDCKII-WT; Pglycoprotein (P-gp); multidrug resistance protein (MRP); breast cancer resistance protein (BCRP); Lopinavir (LVR); uptake; transport; permeability; efflux ratio (ER)

Mechanisms of Reduced Maternal and Fetal Lopinavir Exposure in a Rat Model of Gestational Diabetes

“…More specifically, it has been proposed that Cyp3a drastically lowers LPV concentrations so as to prevent the saturation of Mdr1-mediated hepatobiliary excretion. Agarwal et al (2007) provided evidence supporting the in vitro saturability of Mdr1 by LPV when they observed transporter activity at low LPV concentrations (0.5 M) but not at high LPV concentrations (5-25 M). Data from our study indicate that LPV metabolites also could contribute to Mdr1 saturation.…”

“…LPV is a substrate for the multidrug resistance protein 1 (MDR1) drug efflux transporter (Woodahl et al, 2005;Agarwal et al, 2007) and is metabolized extensively by cytochrome P450 3A4 (CYP3A4) (van Waterschoot et al, 2010). In the liver, MDR1 facilitates LPV's hepatobiliary excretion, whereas CYP3A4 converts it to increasingly hydrophilic metabolites (Fig.…”

The Dual Role of Pharmacogenetics in HIV Treatment: Mutations and Polymorphisms Regulating Antiretroviral Drug Resistance and Disposition

“…OATP2B1 Kumar et al, 1999;Agarwal et al, 2007;Janneh et al, 2007;Weiss et al, 2007a;Hartkoorn et al, 2010;Kis et al, 2010;van Waterschoot et al, 2010 Nelfinavir CYP3A4/5 CYP2C19 ABCB1 ABCC1 ABCG2 OCT1 OCT2 OATP2B1 Kim et al, 1998b;Choo et al, 2000;Zhang et al, 2000;Baede-van Dijk et al, 2001;Jones et al, 2001a;Jones et al, 2001b;Gupta et al, 2004;Hirani et al, 2004;Salama et al, 2005;Kaddoumi et al, 2007;Weiss et al, 2007a;Hirt et al, 2008;Jung et al, 2008;Kis et al, 2010 Ritonavir CYP3A4/5 ABCB1 ABCC1 ABCC2 ABCG2 OCT1 OCT2 OATP2B1 Kumar et al, 1996;Koudriakova et al, 1998;Lee et al, 1998;Zhang et al, 2000;Jones et al, 2001a;Jones et al, 2001b;van der Sandt et al, 2001;Huisman et al, 2002;Gupta et al, 2004;Jung et al, 2008;Zastre et al, 2009;Kis et al, 2010 Saquinavir CYP3A4/5 ABCB1 ABCC1 ABCC2 ABCG2 OATP1A2 OATP1A3 OATP1B1 Fitzsimmons and Collins, 1997;Kim et al, 1998a,b;Kupferschmidt et al, 1998;Lee et al, 1998;Srinivas et al, 1998;…”

MRP2 Inhibition by HIV Protease Inhibitors in Rat and Human Hepatocytes: A Quantitative Confocal Microscopy Study

“…This 'class of most potent' MRP2 inhibitors is largely in agreement with the data obtained by Ye et al, reporting at least 60% inhibition of MRP2 with the conventional method for lopinavir, saquinavir and atazanavir. For the same HIV PI (and including tipranavir), clear evidence was found in literature that they inhibit MRP1 and/or MRP2 (Agarwal et al 2007;Janneh et al 2008;Bierman et al 2010). Taken Figure 1).…”