Simultaneous Assessment of Transporter-Mediated Drug–Drug Interactions Using a Probe Drug Cocktail in Cynomolgus Monkey

Abstract

We aim to establish an in vivo preclinical model to enable simultaneous assessment of inhibition potential of an investigational drug on clinically relevant drug transporters, organic anion-transporting polypeptide (OATP)1B, breast cancer resistance protein (BCRP), P-glycoprotein (P-gp), and organic anion transporter (OAT)3. Pharmacokinetics of substrate cocktail consisting of pitavastatin (OATP1B substrate), rosuvastatin (OATP1B/BCRP/OAT3), sulfasalazine (BCRP), and talinolol (P-gp) were obtained in cynomolgus monkey-alone or in combination with transporter inhibitors. Single-dose rifampicin (30 mg/kg) significantly ( < 0.01) increased the plasma exposure of all four drugs, with a marked effect on pitavastatin and rosuvastatin [area under the plasma concentration-time curve (AUC) ratio ∼21-39]. Elacridar, BCRP/P-gp inhibitor, increased the AUC of sulfasalazine, talinolol, as well as rosuvastatin and pitavastatin. An OAT1/3 inhibitor (probenecid) significantly ( < 0.05) impacted the renal clearance of rosuvastatin (∼8-fold). In vitro, rifampicin (10 M) inhibited uptake of pitavastatin, rosuvastatin, and sulfasalazine by monkey and human primary hepatocytes. Transport studies using membrane vesicles suggested that all probe substrates, except talinolol, are transported by cynoBCRP, whereas talinolol is a cynoP-gp substrate. Elacridar and rifampicin inhibited both cynoBCRP and cynoP-gp in vitro, indicating potential for in vivo intestinal efflux inhibition. In conclusion, a probe substrate cocktail was validated to simultaneously evaluate perpetrator impact on multiple clinically relevant transporters using the cynomolgus monkey. The results support the use of the cynomolgus monkey as a model that could enable drug-drug interaction risk assessment, before advancing a new molecular entity into clinical development, as well as providing mechanistic insights on transporter-mediated interactions.

Physiologically‐Based Pharmacokinetic Models for Evaluating Membrane Transporter Mediated Drug–Drug Interactions: Current Capabilities, Case Studies, Future Opportunities, and Recommendations

“…Hence, another OAT1/3 probe may be more reliable ( Figure b ). Recently, a substrate cocktail consisting of pitavastatin (OATP1B), rosuvastatin (OATP1B/BCRP/OAT3), sulfasalazine (BCRP), and talinolol (P‐gp) was evaluated in cynomolgus monkey …”

In vitro and in vivo methods to assess pharmacokinetic drug– drug interactions in drug discovery and development

“…Clinical DDI studies with cocktail substrates reduce the number of clinical trials, speed up drug development processes and enable better ethical compliance (Trueck et al, ). Similar cocktail approaches have been reported in human with micro‐dosing and in cynomolgus monkeys for both major CYPs and transporters (Kosa et al, ; Prueksaritanont et al, ). For transporter‐mediated DDI, cynomolgus monkey has been found to be a good model for prediction of OATP1B, BCRP, P‐gp, and OAT3 mediated DDI in humans (Kosa et al, ).…”

“…Animal models typically cannot reliably predict human DDIs, with the exception of a few reports on transporter DDIs. It has been shown that cynomolgus monkey may be a viable model to predict the activities and DDIs of major drug transporters in humans such as OATP1Bs, OAT3, P‐gp, and BCRP (Kosa et al, ). Both static or dynamic models are available for human DDI prediction.…”

Resveratrol glucuronidation in vitro: potential implications of inhibition by probenecid

“…In vitro work suggests that probenecid has lower inhibitory activity vs resveratrol sulfate formation compared with the glucuronide conjugation pathway . Also, probenecid minimally inhibits transport mediated by P‐gp …”

“…[70] Also, probenecid minimally inhibits transport mediated by P-gp. [71][72][73] The observed in vitro inhibition of glucuronidation by probenecid translates to clinical DDIs, in which coadministration of usual therapeutic doses of probenecid causes reduced clearance and increased plasma concentrations of a number of substrate drugs cleared mainly by glucuronide conjugation. Examples of affected substrates include paracetamol (acetaminophen), lorazepam, oxazepam, temazepam, diflunisal, zomepirac, zidovudine, and naproxen.…”