Quantitation of Plasma Membrane Drug Transporters in Kidney Tissue and Cell Lines Using a Novel Proteomic Approach Enabled a Prospective Prediction of Metformin Disposition.

Autor:	Kikuchi R; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Chiou WJ; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Durbin KR; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Savaryn JP; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Ma J; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Emami Riedmaier A; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., de Morais SM; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Jenkins GJ; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois., Bow DAJ; Drug Metabolism and Pharmacokinetics, AbbVie Inc., North Chicago, Illinois daniel.bow@abbvie.com.
Jazyk:	angličtina
Zdroj:	Drug metabolism and disposition: the biological fate of chemicals [Drug Metab Dispos] 2021 Oct; Vol. 49 (10), pp. 938-946. Date of Electronic Publication: 2021 Jul 30.
DOI:	10.1124/dmd.121.000487
Abstrakt:	The successful prospective incorporation of in vitro transporter kinetics in physiologically based pharmacokinetic (PBPK) models to describe drug disposition remains challenging. Although determination of scaling factors to extrapolate in vitro to in vivo transporter kinetics has been facilitated by quantitative proteomics, no robust assessment comparing membrane recoveries between different cells/tissues has been made. HEK293 cells overexpressing OCT2, MATE1, and MATE2K or human kidney cortex were homogenized and centrifuged to obtain the total membrane fractions, which were subsequently subjected to liquid-liquid extraction followed by centrifugation and precipitation to isolate plasma membrane fractions. Plasma membrane recoveries determined by quantitation of the marker Na + /K + -ATPase in lysate and plasma membrane fractions were ≤20% but within 3-fold across different cells and tissues. A separate study demonstrated that recoveries are comparable between basolateral and apical membranes of renal proximal tubules, as measured by Na + /K + -ATPase and γ -glutamyl transpeptidase 1, respectively. The plasma membrane expression of OCT2, MATE1, and MATE2K was quantified and relative expression factors (REFs) were determined as the ratio between the tissue and cell concentrations. Corrections using plasma membrane recovery had minimal impact on REF values (<2-fold). In vitro transporter kinetics of metformin were extrapolated to in vivo using the corresponding REFs in a PBPK model. The simulated metformin exposures were within 2-fold of clinical exposure. These results demonstrate that transporter REFs based on plasma membrane expression enable a prediction of transporter-mediated drug disposition. Such REFs may be estimated without the correction of plasma membrane recovery when the same procedure is applied between different matrices. SIGNIFICANCE STATEMENT: Transporter REFs based on plasma membrane expression enable in vitro-in vivo extrapolation of transporter kinetics. Plasma membrane recoveries as determined by the quantification of sodium-potassium adenosine triphosphatase were comparable between the in vitro and in vivo systems used in the present study, and therefore had minimal impact on the transporter REF values. (Copyright © 2021 by The American Society for Pharmacology and Experimental Therapeutics.)
Databáze:	MEDLINE
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