title: Evaluation of multiple in vitro systems for assessment of CYP3A4 induction in drug discovery: human hepatocytes, pregnane X receptor reporter gene, and Fa2N-4 and HepaRG cells.
abstract: Prototypic CYP3A4 inducers were tested in a pregnane X receptor (PXR) reporter gene assay, Fa2N-4 cells, HepaRG cells, and primary human hepatocytes, along with negative controls, using CYP3A4 mRNA and activity endpoints, where appropriate. Over half of the compounds tested (14 of 24) were identified as time-dependent inhibitors of CYP3A4 and high mRNA/activity ratios (>10) were consistent with CYP3A4 time-dependent inhibition for compounds such as troleandomycin, ritonavir, and verapamil. Induction response was compared between two human donors; there was an excellent correlation in the EC(50) estimates (r(2) = 0.89, p < 0.001), and a weak but statistically significant correlation was noted for maximum observed induction at an optimum concentration (E(max)) (r(2) = 0.38, p = 0.001). E(max) and EC(50) estimates determined from the PXR reporter gene assay and Fa2N-4 and HepaRG cells were compared with those from hepatocytes. Overall, EC(50) values generated using hepatocytes agreed with those generated in the PXR reporter gene assay (r(2) = 0.85, p < 0.001) and Fa2N-4 (r(2) = 0.65, p < 0.001) and HepaRG (r(2) = 0.99, p < 0.001) cells. However, E(max) values generated in hepatocytes were only significantly correlated to those determined in Fa2N-4 (r(2) = 0.33, p = 0.005) and HepaRG cells (r(2) = 0.79, p < 0.001). "Gold standard" cytochrome P450 induction data can be generated using primary human hepatocytes, but a restricted, erratic supply and interdonor variability somewhat restrict routine application within a drug discovery setting. HepaRG cells are a valuable recent addition to the armory of in vitro tools for assessing CYP3A4 induction and seem to be an excellent surrogate of primary cells.
authors.affiliation: Discovery DMPK, AstraZeneca Research and Development, Charnwood, UK. dermot.f.mcginnity@astrazeneca.com
authors.forename: Dermot F
authors.initials: DF
authors.lastname: McGinnity
authors.forename: George
authors.initials: G
authors.lastname: Zhang
authors.forename: Jane R
authors.initials: JR
authors.lastname: Kenny
authors.forename: Geraldine A
authors.initials: GA
authors.lastname: Hamilton
authors.forename: Sara
authors.initials: S
authors.lastname: Otmani
authors.forename: Karen R
authors.initials: KR
authors.lastname: Stams
authors.forename: Suzzette
authors.initials: S
authors.lastname: Haney
authors.forename: Patrick
authors.initials: P
authors.lastname: Brassil
authors.forename: David M
authors.initials: DM
authors.lastname: Stresser
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Riley
mesh_terms: D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis*; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D004347:Drug Interactions; D004790:Enzyme Induction / Q000187:drug effects; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D004364:Pharmaceutical Preparations; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D004364:Pharmaceutical Preparations; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human