Drug Metabolism

Nuclear Receptor Activation


  • The primary mechanism of cytochrome P450 induction is via increased gene transcription which typically occurs through nuclear receptor activation.
  • The most common nuclear receptors involved in the induction of drug metabolizing enzymes include the pregnane X receptor (PXR), the aryl hydrocarbon receptor (AhR), and the constitutive androstane receptor (CAR) which are known to regulate CYP3A4, CYP1A2 and CYP2B6, respectively.
  • An industry survey of current practices and recommendations (1Chu et al., (2009) Drug Metab Dispos 37; 1339) indicates 64% of survey respondents routinely use nuclear receptor transactivation assays to assess the potential of test compounds to cause enzyme induction.
  • Cyprotex can evaluate PXR and AhR nuclear receptor activation utilizing stably-transfected human hepatoma cell lines (DPX2™ for PXR and 1A2-DRE™ for AhR) and a luciferase reporter gene assay.
  • In addition, we can also evaluate CAR-3 using INDIGO's proprietary CAR3 Reporter Cells (mammalian cells engineered to provide high-level expression of human CAR3).


PXR & AhR Nuclear Receptor Activation Assay Protocol

CAR-3 Nuclear Receptor Activation Assay Protocol


Data from Cyprotex's Nuclear Receptor Activation Assay

Known activator of PXR was selected and screened in the PXR nuclear receptor activation assay. Data generated were compared to those published in the literature.

Web graphics Cyprotex 231024 01

Figure 1
PXR nuclear receptor activation by rifampicin.
Data show the mean fold activation relative to the vehicle control. Error bars represent the standard deviation of 2 replicate incubations.

Data analysis of the CAR3 nuclear receptor activation assay:

Transcriptional activation is monitored by luminescence. Data are expressed as fold activation relative to the vehicle control, following normalization of luciferase activity against cell viability. The use of 5 or more doses of test article and positive control allows for the derivation of EC50 and Emax values from nonlinear regression analysis of the log dose‑response curves. These outputs may not be determined with test articles exhibiting atypical dose‑response curves or with poorly defined parameters. The average observed fold activation is calculated as a percentage of CITCO (2.5 µM) to allow for inter‑day comparison and ranking of test articles, using the equation below:

of CITCO (2.5 μM) =(Fold activation of test article)(Fold activation of CITCO (2.5 μM))x 100


1) Chu V et al., (2009) Drug Metab Dispos 37; 1339-1354
2) Yueh M-F et al., (2005) Drug Metab Dispos 33; 38-48


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Cyprotex enables and enhances the prediction of human exposure, clinical efficacy and toxicological outcome of a drug or chemical. By combining quality data from robust in vitro methods with contemporary in silico technology, we add value, context and relevance to the ADME-Tox data supplied to our partners in the pharmaceutical or chemical industries.