Pharmacelsus – Success Story 4

Pharmacelsus as the lead lab in a formal ECVAM Validation trial on CytochromeP450 induction

Challenge

Pharmacelsus was selected to be the lead lab in an international metabolism and toxicology validation study which became part of a formal ECVAM validation trial entitled “Multi-study validation trial for cytochromeP450 induction providing a standard for human hepatic metabolism and toxicity using HepaRG^® cell line and cryopreserved hepatocytes”.

Idea

A validation trial was initiated aiming to assess the reliability and relevance with a challenging set of chemicals for which high quality in vivo data are available.

First Steps

The induction protocol was optimised for HepaRG^® cells in a 96-well-plate format using specific CYP1A2, CYP2B6, CYP2C9, and CYP3A4 probe substrates. Conversion was measured by LC/MS using a cocktail approach (see also Pharmacelsus Success Story 3).

Solution

By combining a multi-well screening format with n-in-one incubation and LC/MS cocktail analysis, a state-of-the-art high content screening platform for CYP induction was set up. Applying that protocol, HepaRG^® cells can be used as cost effective and truly predictive surrogate for human hepatic CYP induction studies.

Results

Pharmacelsus confirmed that cryopreserved HepaRG^® were advantageous over fresh HepaRG^® cells for CYP induction studies. Furthermore we demonstrated within-lab reproducibility and reliability and successfully transferred the method to two prototypical naïve labs.

Perhaps it is not about getting the cheapest CRO, but about getting results from a CRO, who is on the front lines of assay development!