A Clinical Cassette Dosing Study for Evaluating the Contribution of Hepatic OATPs and CYP3A to Drug-Drug Interactions



To demonstrate the relative importance of organic anion-transporting polypeptides (OATPs) and cytochrome P450 3A (CYP3A) in the hepatic elimination of substrate drugs.


A cocktail of subtherapeutic doses of bosentan, repaglinide, clarithromycin, darunavir, simeprevir, and midazolam (CYP3A probe) was administered orally to eight healthy volunteers. Rifampicin (OATP inhibitor; 600 mg, p.o.) and itraconazole (CYP3A inhibitor; 200 mg, i.v.) were coadministered with the cocktail in the second and third phases, respectively. Based on the extended clearance concept, in vivo β values (fraction of metabolism plus biliary excretion among all the intracellular fates of drugs including basolateral efflux) and Rdif values (ratio of diffusional uptake to active uptake) were estimated.


Rifampicin increased plasma AUCs of bosentan (×3.2), repaglinide (×1.9), clarithromycin (×1.9) and simeprevir (×7.2). Itraconazole increased those of clarithromycin (×2.3), simeprevir (×2.2) and midazolam (×3.7), which had relatively small β values. The plasma AUC of bosentan (with relatively large β and small Rdif) was dominated by OATP-mediated uptake. The AUC of simeprevir was also dominated by OATP-mediated uptake because of its small Rdif value.


The DDI study clarified the rate-determining processes of OATP/CYP3A substrates. Our analyses provide valuable information for predicting complex drug–drug interactions involving multiple processes.

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Fig. 1
Fig. 2
Fig. 3



Area under the plasma concentration–time curve


AUC ratio



CLh :

Hepatic clearance

CLint,all :

Overall hepatic intrinsic clearance

CLint,bile :

Intrinsic clearance of biliary excretion

CLint,met :

Intrinsic clearance of hepatic metabolism


Renal clearance


Cytochrome P450


Drug-drug interaction


University of Washington’s metabolism and transporter drug interaction database program


Extended clearance classification system


Extended clearance concept classification system

Fa :

Fraction of the oral dose that enters the gut wall

fB :

Protein unbound fraction in blood

Fg :

Fraction of drug passing on to the portal circulation

Fh :

Hepatic availability

fm,CYP3A :

Contribution of CYP3A to the overall metabolic activity

fOATP1Bs :

Fraction of OATP1Bs-mediated transport in the hepatic intrinsic uptake clearance


Inhibition ratio for CYP3A by itraconazole


Inhibition ratio for OATP1Bs by rifampicin


in vitro-in vivo extrapolation


Na+-taurocholate cotransporting polypeptide


Organic anion transporting polypeptide


Physiologically-based pharmacokinetic


Permeability surface area-product

PSact,eff :

Intrinsic active efflux clearance on sinusoidal membrane

PSact,inf :

Intrinsic active uptake clearance on sinusoidal membrane

PSdif,eff :

Intrinsic efflux clearance by passive diffusion on sinusoidal membrane

PSdif,inf :

Intrinsic influx clearance by passive diffusion on sinusoidal membrane

Qh :

Hepatic blood flow rate

RB :

Blood-to-plasma concentration ratio

Rdif :

Ratio of PSdif,inf to PSact,inf


Sandwich-cultured human hepatocyte




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Acknowledgements and Disclosures

This study was financially supported by Grant-in-Aid for Scientific Research (S) [Grant 24,229,002; TY, KM, YS].

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Corresponding author

Correspondence to Yuichi Sugiyama.

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Wrote Manuscript: Takashi Yoshikado, Kazuya Maeda, Hiroyuki Kusuhara, and Yuichi Sugiyama.

Designed Research: Takashi Yoshikado, Kazuya Maeda, Sawako Furihata, Hiroyuki Kusuhara, Ken-ichi Furihata and Yuichi Sugiyama.

Performed Research: Takashi Yoshikado, Kazuya Maeda, Sawako Furihata, Hanano Terashima, Takeshi Nakayama, Keiko Ishigame, Kazunobu Tsunemoto, Hiroyuki Kusuhara, Ken-ichi Furihata and Yuichi Sugiyama.

Analyzed Data: Takashi Yoshikado, Kazuya Maeda, Hiroyuki Kusuhara, and Yuichi Sugiyama.

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Yoshikado, T., Maeda, K., Furihata, S. et al. A Clinical Cassette Dosing Study for Evaluating the Contribution of Hepatic OATPs and CYP3A to Drug-Drug Interactions. Pharm Res 34, 1570–1583 (2017). https://doi.org/10.1007/s11095-017-2168-5

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Key words

  • Extended clearance concept
  • Hepatic uptake
  • Metabolism
  • Rate-determining process