Abstract
The liver plays an important role in the clearance of endogenous and exogenous compounds, including drugs. As hepatic uptake is the first step in hepatic clearance, any change in the former process directly affects the overall intrinsic hepatic clearance. Several uptake transporters are expressed on the basolateral membranes of hepatocytes and mediate the hepatic uptake of hydrophilic charged compounds that cannot easily penetrate the plasma membrane. As the substrate specificities of these individual drug transporters are broad and overlap, compounds are often recognized by multiple uptake transporters. Thus, knowledge of the contribution that each transporter makes to the hepatic uptake of a compound is important for predicting the extent to which hepatic uptake clearance will change if the activity of a specific transporter is altered by a genetic polymorphism or a drug–drug interaction. Human cryopreserved hepatocytes are now commercially available and can be used for studying hepatic uptake clearance. In this chapter, we describe a method for using isolated hepatocytes to estimate the in vivo uptake clearance of compounds and the quantitative contribution of each uptake transporter to the overall hepatic uptake of anionic compounds.
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References
Giacomini, K.M. and Sugiyama, Y. (2005) Membrane transporters and drug response, in Goodman & Gilman’s the Pharmacological Basis of Therapeutics, 11th ed. (Brunton, L.L., Lazo, J.S., and Parker, K.L., eds.), McGraw-Hill, New York, pp. 41–70.
Shitara, Y., Horie, T., and Sugiyama, Y. (2006) Transporters as a determinant of drug clearance and tissue distribution. Eur. J. Pharm. Sci. 27, 425–446.
Regazzi, M.B., Iacona, I., Campana, C., Raddato, V., Lesi, C., Perani, G., Gavazzi, A., and Vigano, M. (1993) Altered disposition of pravastatin following concomitant drug therapy with cyclosporin A in transplant recipients. Transplant. Proc. 25, 2732–2734.
Asberg, A., Hartmann, A., Fjeldsa, E., Bergan, S., and Holdaas, H. (2001) Bilateral pharmacokinetic interaction between cyclosporine A and atorvastatin in renal transplant recipients. Am. J. Transplant. 1, 382–386.
Ichimaru, N., Takahara, S., Kokado, Y., Wang, J.D., Hatori, M., Kameoka, H., Inoue, T., and Okuyama, A. (2001) Changes in lipid metabolism and effect of simvastatin in renal transplant recipients induced by cyclosporine or tacrolimus. Atherosclerosis 158, 417–423.
Mazzu, A.L., Lasseter, K.C., Shamblen, E.C., Agarwal, V., Lettieri, J., and Sundaresen, P. (2000) Itraconazole alters the pharmacokinetics of atorvastatin to a greater extent than either cerivastatin or pravastatin. Clin. Pharmacol. Ther. 68, 391–400.
Neuvonen, P.J., Kantola, T., and Kivisto, K.T. (1998) Simvastatin but not pravastatin is very susceptible to interaction with the CYP3A4 inhibitor itraconazole. Clin. Pharmacol. Ther. 63, 332–341.
Maeda, K. and Sugiyama, Y. (2008) Impact of genetic polymorphisms of transporters on the pharmacokinetic, pharmacodynamic and toxicological properties of anionic drugs. Drug Metab. Pharmacokinet. 23, 223–235.
Kiyotani, K., Mushiroda, T., Kubo, M., Zembutsu, H., Sugiyama, Y., and Nakamura, Y. (2008) Association of genetic polymorphisms in SLCO1B3 and ABCC2 with docetaxel-induced leukopenia. Cancer Sci. 99, 967–972.
Noe, J., Portmann, R., Brun, M.E., and Funk, C. (2007) Substrate-dependent drug-drug interactions between gemfibrozil, fluvastatin and other organic anion-transporting peptide (OATP) substrates on OATP1B1, OATP2B1, and OATP1B3. Drug Metab. Dispos. 35, 1308–1314.
Link, E., Parish, S., Armitage, J., Bowman, L., Heath, S., Matsuda, F., Gut, I., Lathrop, M., and Collins, R. (2008) SLCO1B1 variants and statin-induced myopathy – a genomewide study. N. Engl. J. Med. 359, 789–799.
Hirano, M., Maeda, K., Shitara, Y., and Sugiyama, Y. (2004) Contribution of OATP2 (OATP1B1) and OATP8 (OATP1B3) to the hepatic uptake of pitavastatin in humans. J. Pharmacol. Exp. Ther. 311, 139–146.
Hirano, M., Maeda, K., Shitara, Y., and Sugiyama, Y. (2006) Drug-drug interaction between pitavastatin and various drugs via OATP1B1. Drug Metab. Dispos. 34, 1229–1236.
Crespi, C.L. and Penman, B.W. (1997) Use of cDNA-expressed human cytochrome P450 enzymes to study potential drug-drug interactions. Adv. Pharmacol. 43, 171–188.
Kouzuki, H., Suzuki, H., Ito, K., Ohashi, R., and Sugiyama, Y. (1998) Contribution of sodium taurocholate co-transporting polypeptide to the uptake of its possible substrates into rat hepatocytes. J. Pharmacol. Exp. Ther. 286, 1043–1050.
Kouzuki, H., Suzuki, H., Ito, K., Ohashi, R., and Sugiyama, Y. (1999) Contribution of organic anion transporting polypeptide to uptake of its possible substrates into rat hepatocytes. J. Pharmacol. Exp. Ther. 288, 627–634.
Miyauchi, S., Sawada, Y., Iga, T., Hanano, M., and Sugiyama, Y. (1993) Comparison of the hepatic uptake clearances of fifteen drugs with a wide range of membrane permeabilities in isolated rat hepatocytes and perfused rat livers. Pharm. Res. 10, 434–440.
Kato, Y., Akhteruzzaman, S., Hisaka, A., and Sugiyama, Y. (1999) Hepatobiliary transport governs overall elimination of peptidic endothelin antagonists in rats. J. Pharmacol. Exp. Ther. 288, 568–574.
Watanabe, T., Maeda, K., Kondo, T., Nakayama, H., Horita, S., Kusuhara, H., and Sugiyama, Y. (2009) Prediction of the hepatic and renal clearance of transporter substrates in rats using in vitro uptake experiments. Drug Metab. Dispos. 37, 1471–1479.
Kitamura, S., Maeda, K., Wang, Y., and Sugiyama, Y. (2008) Involvement of multiple transporters in the hepatobiliary transport of rosuvastatin. Drug Metab. Dispos. 36, 2014–2023.
Ishiguro, N., Maeda, K., Kishimoto, W., Saito, A., Harada, A., Ebner, T., Roth, W., Igarashi, T., and Sugiyama, Y. (2006) Predominant contribution of OATP1B3 to the hepatic uptake of telmisartan, an angiotensin II receptor antagonist, in humans. Drug Metab. Dispos. 34, 1109–1115.
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Maeda, K., Sugiyama, Y. (2010). The Use of Hepatocytes to Investigate Drug Uptake Transporters. In: Maurel, P. (eds) Hepatocytes. Methods in Molecular Biology, vol 640. Humana Press. https://doi.org/10.1007/978-1-60761-688-7_18
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DOI: https://doi.org/10.1007/978-1-60761-688-7_18
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