Pre‐incubation with cyclosporine A potentiates its inhibitory effects on pitavastatin uptake mediated by recombinantly expressed cynomolgus monkey hepatic organic anion transporting polypeptide

Abstract

Cyclosporine A, an inhibitor of hepatic organic anion transporting polypeptides (OATPs), reportedly increased plasma concentrations of probe substrates, although its maximum unbound blood concentrations were lower than the experimental half-maximal inhibitory (IC ) concentrations. Pre-incubation with cyclosporine A in vitro before simultaneous incubation with probes has been reported to potentiate its inhibitory effects on recombinant human OATP-mediated probe uptake. In the present study, the effects of cyclosporine A and rifampicin on recombinant cynomolgus monkey OATP-mediated pitavastatin uptake were investigated in pre- and simultaneous incubation systems. Pre-incubation with cyclosporine A, but not with rifampicin, decreased the apparent IC values on recombinant cynomolgus monkey OATP1B1- and OATP1B3-mediated pitavastatin uptake. Application of the co-incubated IC values toward R values (1 + [unbound inhibitor] /inhibition constant) in static models, 1.1 in monkeys and 1.3 in humans, for recombinant cynomolgus monkey and human OATP1B1-mediated pitavastatin uptake might result in the poor prediction of drug interaction magnitudes. In contrast, the lowered IC values after pre-incubation with cyclosporine A provided better prediction with R values of 3.9 for monkeys and 2.7 for humans when the estimated maximum cyclosporine A concentrations at the inlet to the liver were used. These results suggest that the enhanced inhibitory potential of perpetrator medicines by pre-incubation on cynomolgus monkey OATP-mediated pitavastatin uptake in vitro could be of value for the precise estimation of drug interaction magnitudes in silico, in accordance with the findings from pre-administration of inhibitors on pitavastatin pharmacokinetics validated in monkeys. Copyright © 2016 John Wiley & Sons, Ltd.

Gaining Mechanistic Insight Into Coproporphyrin I as Endogenous Biomarker for OATP1B‐Mediated Drug–Drug Interactions Using Population Pharmacokinetic Modeling and Simulation

“…The uncertainty in the literature reported in vitro inhibition parameters (30‐fold range in RIF IC 50 , Table S5 ) and substrate‐dependent inhibition evident for OATP1B1 makes the translational aspect of OATP1B DDIs challenging, even for prototypical inhibitors like RIF. In addition, methodological considerations, e.g., preincubation with an inhibitor, have been reported to increase the potency of OATP1B inhibition in vitro , as recently acknowledged by the US Food and Drug Administration (FDA) DDI guidance . RIF model‐based in vivo Ki values were 3–4‐fold lower compared with in vitro parameters obtained in human hepatocytes with the corresponding probes and following preincubation with inhibitor ( Figure b ).…”

Mechanistic Evaluation of the Complex Drug-Drug Interactions of Maraviroc: Contribution of Cytochrome P450 3A, P-Glycoprotein and Organic Anion Transporting Polypeptide 1B1

“…The empirical correction of the in vitro transporter K i value may add some degree of uncertainty to the interpretation of quantitative contribution of the individual mechanisms to the noted DDIs. The potential TDI for transporters, such as cyclosporin A, could be one possibility (Takahashi et al, 2016;Shitara and Sugiyama, 2017). Nevertheless, our crossvalidation approach to verify individual interaction mechanisms of TVR using clinical data from probe substrates (e.g., midazolam for CYP3A, digoxin for P-gp, and atorvastatin for OATP1B1) provided rationale for such empirical corrections to the less understood in vitro-in vivo disconnect in transporter inhibition potency.…”

Functional characterization for polymorphic organic anion transporting polypeptides (OATP/SLCO1B1, 1B3, 2B1) of monkeys recombinantly expressed with various OATP probes

“…For telmisartan and simeprevir, HBSS containing 0.3% bovine serum albumin was used. In our previous study (Takahashi et al, 2016), the uptake of [ 3 H]pitavastatin for cynomolgus monkey OATP1B1-and OATP1B3-expressing cells increased linearly up to 3 min during the 5 min incubation. Then, uptake was initiated by replacing the medium with HBSS containing substrate compounds at 37°C for 2 min in kinetic assays or 10 min for capacity assays in this study.…”

“…The uptake study was carried out as described previously with the following modifications using human embryonic kidney 293 (HEK293) cells transiently expressing recombinant cynomolgus monkey OATP1B1, OATP1B3 and OATP2B1, and OATP1B1 variants, along with the control HEK293 cells (mock cells) (Takahashi et al, ). The cells were transfected using the Lipofectamine® 2000 protocol (Thermo Fisher Scientific Inc. Rockford, IL) with the pcDNA5/FRT vector containing the cynomolgus monkey OATP1B1, OATP1B3, OATP2B1 or OATP1B1 variant, or pcDNA5/FRT (for mock) vector, according to the manufacturer's protocols.…”

“…The uptake study was carried out as described previously with the following modifications using human embryonic kidney 293 (HEK293) cells transiently expressing recombinant cynomolgus monkey OATP1B1, OATP1B3 and OATP2B1, and OATP1B1 variants, along with the control HEK293 cells (mock cells) (Takahashi et al, 2016).…”