Metabolite Formation Kinetics and Intrinsic Clearance of Phenacetin, Tolbutamide, Alprazolam, and Midazolam in Adenoviral Cytochrome P450-Transfected HepG2 Cells and Comparison with Hepatocytes and In Vivo

Abstract

Cryopreserved human hepatocytes and other in vitro systems often underpredict in vivo intrinsic clearance (CL int)

Effect of Albumin on Human Liver Microsomal and Recombinant CYP1A2 Activities: Impact on In Vitro-In Vivo Extrapolation of Drug Clearance

“…In the absence of BSA, the respective K m values of the high-affinity component of PHEN O-deethylation by HLM and rCYP1A2 (11.4 -67.8 and 13.6 M) were similar to previously published ranges (9 -38.4 and 12.3-30.9 M) (Gillam and Reilly, 1988;Tassaneeyakul et al, 1993;Venkatakrishnan et al, 1998;Kobayashi et al, 1999;Polasek et al, 2006b;Donato et al, 2010). Likewise, K m values of the high-affinity component of LID N-deethylation by HLM (135-246 M) were similar to data reported by Wang et al (2000).…”

Prediction of Human Metabolic Clearance from In Vitro Systems: Retrospective Analysis and Prospective View

“…Hence, there is a need for novel cellular systems that are devoid of this limitation and display the advantages of both systems. Cell lines which are genetically engineered to provide activities comparable to what is believed to be occurring in vivo should be the aspiration (14,26). The large database of in vivo numbers enclosed here indicates the range that is required and provides the necessary in vivo correlates to allow a top-down assessment of any novel in vitro system for clearance prediction.…”

Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME

“…Recombinant adenoviruses rapidly and efficiently infect hepatic cell lines, and almost 100 % of the cells can express functional levels of the transgene after a short exposure to the virus (Castell et al 1997). This technology has facilitated the generation of transient metabolically competent cells after transduction with recombinant-defective adenoviral vectors encoding for CYP genes (Castell et al 1997; Bai and Cederbaum 2004; Naiki et al 2004; Hosomi et al 2011; Donato et al 2010). A major characteristic of this approach is that functional levels of CYP are easily modulated as a function of the number of infecting virus particles (Fig.…”

“…Among several hepatoma cell lines (HepG2, Hep3B, HLE, HLF, Huh6, Huh7 and Fa2N4 cells) efficiently infected with adenovirus vector harboring CYP3A4, HepG2 cells showed the highest CYP3A4 protein expression and corresponding testosterone 6β-hydroxylase activity (Hosomi et al 2011). The use of adenoviral strategy to confer CYP activities to HepG2 cells has been extensively reported (Vignati et al 2005; Donato et al 2010; Aoyama et al 2009). This cell system has been proposed as a new in vitro tool for metabolism-mediated toxicity and clearance prediction of drugs metabolized by CYPs (Vignati et al 2005; Donato et al 2010).…”