Cynomolgus Monkey as a Clinically Relevant Model to Study Transport Involving Renal Organic Cation Transporters: In Vitro and In Vivo Evaluation

Abstract

Organic cation transporter (OCT) 2, multidrug and toxin extrusion protein (MATE) 1, and MATE2K mediate the renal secretion of various cationic drugs and can serve as the loci of drug-drug interactions (DDI). To support the evaluation of cynomolgus monkey as a surrogate model for studying human organic cation transporters, monkey genes were cloned and shown to have a high degree of amino acid sequence identity versus their human counterparts (93.7, 94.7, and 95.4% for OCT2, MATE1, and MATE2K, respectively). Subsequently, the three transporters were individually stably expressed in human embryonic kidney (HEK) 293 cells and their properties (substrate selectivity, time course, pH dependence, and kinetics) were found to be comparable to the corresponding human form. For example, six known human cation transporter inhibitors, including pyrimethamine (PYR), showed generally similar IC 50 values against the monkey transporters (within sixfold). Consistent with the in vitro inhibition of metformin (MFM) transport by PYR (IC 50 for cynomolgus OCT2, MATE1, and MATE2K; 1.2 6 0.38, 0.17 6 0.04, and 0.25 6 0.04 mM, respectively), intravenous pretreatment of monkeys with PYR (0.5 mg/kg) decreased the clearance (54 6 9%) and increased in the area under the plasma concentration-time curve of MFM (AUC ratio versus control = 2.23; 90% confidence interval of 1.57 to 3.17). These findings suggest that the cynomolgus monkey may have some utility in support of in vitroin vivo extrapolations (IVIVEs) involving the inhibition of renal OCT2 and MATEs. In turn, cynomolgus monkey-enabled IVIVEs may inform human DDI risk assessment. IntroductionIt is widely appreciated that renal elimination of cations, drug, toxin, or endogenous metabolite related, is achieved not only by glomerular filtration but also by active transport processes that facilitate their tubular secretion and reabsorption. Indeed, the mechanisms governing the renal elimination of organic cations have been studied in detail, and the transporters facilitating their tubular uptake and extrusion have been identified (Okuda et al., 1996;Otsuka et al., 2005;Masuda et al., 2006).Secretion of organic cations in renal proximal tubules involves at least two distinct transporters, located in the basolateral and apical membranes of proximal tubule cells (Motohashi et al., 2002Masuda et al., 2006). Specifically, organic cations, like metformin (MFM), 1-methyl-4-phenylpyridinium (MPP + ), tetraethylammonium (TEA), and cimetidine (CMD), are taken up from the circulation by organic cation transporter 2 (OCT2) expressed on the basolateral domain of renal proximal tubular cells. In turn, uptake is followed by efflux into the tubular fluid by multidrug and toxin extrusion protein (MATE) 1 and MATE2K expressed on the apical domain of the same cells. Therefore, OCT2 and MATEs function coordinately to mediate vectorial transport of certain cationic drugs, from blood to tubular fluid, which represents the tubular secretion clearance component of total renal clearance. Perturbation of c...

Mechanistic in vitro studies confirm that inhibition of the renal apical efflux transporter multidrug and toxin extrusion (MATE) 1, and not altered absorption, underlies the increased metformin exposure observed in clinical interactions with cimetidine, trimethoprim or pyrimethamine

“…Determined IC 50 (K i ) values (Table 2) versus OCT2, MATE1 and MATE2-K were in reasonable concordance with literature values and were much lower (35-to 170-fold; Fig. 1) for MATEs versus OCT2 (Kusuhara et al 2011;Ito et al 2012;M€ uller et al 2015;Shen et al, 2016). However, only inhibition constants for OCT2 and MATE1 were subsequently incorporated into mechanistic equations for predicting metformin AUC increases as the study of Prasad et al (2016) determined that these, and not MATE2-K, are the major cationic transporter proteins expressed in human renal proximal tubules; representing 26% and 18% of renal transporter protein.…”

“…; Müller et al. ; Shen et al., ). However, only inhibition constants for OCT2 and MATE1 were subsequently incorporated into mechanistic equations for predicting metformin AUC increases as the study of Prasad et al.…”

Coproporphyrins in Plasma and Urine Can Be Appropriate Clinical Biomarkers to Recapitulate Drug-Drug Interactions Mediated by Organic Anion Transporting Polypeptide Inhibition

“…The results suggested that CPs could be potential endogenous biomarkers of OATP activity in vivo (Benz-de Bretagne et al, 2011;van de Steeg et al, 2012). Indeed, the plasma concentrations of both CP-I and CP-III in cynomolgus monkeys were markedly increased following administration of OATP inhibitors cyclosporine A (100 mg/kg oral) or RIF (15 mg/kg oral) (Shen et al, 2015(Shen et al, , 2016b. The observations in monkey could explain the clinical findings that cyclosporine A-or RIF-induced porphyria in patients is likely due to the inhibition of OATP transporters (Millar, 1980;Hivnor et al, 2003).…”

Globotriaosylsphingosine induces oxidative DNA damage in cultured kidney cells

“…In this work we chose a human embryonic kidney cell‐line 293 T (HEK‐293 T) to evaluate the effects on DNA damage of the latest described potential biomarker for FD – lyso‐Gb3 – regarding to oxidative damage. HEK‐293 and HEK‐293 T cells have been used as a model of epithelial kidney cell‐line in renal physiology studies . To reproduce pathological conditions, all experiments were done using lyso‐Gb3 in concentrations found in plasma of Fabry patients: 10, 50 and 100 nM .…”

“…HEK-293 and HEK-293 T cells have been used as a model of epithelial kidney cell-line in renal physiology studies. [14][15][16][17] To reproduce pathological conditions, all experiments were done using lyso-Gb3 in concentrations found in plasma of Fabry patients: 10, 50 and 100 nM. 5,[7][8][9][10] To the best of our knowledge, this is the first study focusing on these effects of lyso-Gb3.…”