Quantitative In Vitro-to-In Vivo Extrapolation: Nominal versus Freely Dissolved Concentration

Discussions are ongoing on which dose metric should be used for quantitative in vitro-to-in vivo extrapolation (QIVIVE) of in vitro bioassay data. The nominal concentration of the test chemicals is most commonly used and easily accessible, while the concentration freely dissolved in the assay medium is considered to better reflect the bioavailable concentration but is tedious to measure. The aim of this study was to elucidate how much QIVIVE results will differ when using either nominal or freely dissolved concentrations. QIVIVE_nom and QIVIVE_free ratios, that is, the ratios of plasma concentrations divided by in vitro effect concentrations, were calculated for 10 pharmaceuticals using previously published nominal and freely dissolved effect concentrations for the activation of the peroxisome proliferator-activated receptor gamma (PPARγ) and the activation of oxidative stress response. The QIVIVE_nom ratios were higher than QIVIVE_free ratios by up to a factor of 60. The risk of in vivo effects was classified as being high or low for four chemicals using the QIVIVE_nom and for three chemicals using QIVIVE_free ratios. Unambiguous classification was possible for nine chemicals by combining the QIVIVE_nom or QIVIVE_free ratios with the respective specificity ratios (SR_nom or SR_free) of the in vitro effect data, which helps to identify whether the specific effect was influenced by cytotoxicity. QIVIVE_free models should be preferred as they account for differences in bioavailability between in vitro and in vivo, but QIVIVE_nom may still be useful for screening the effects of large numbers of chemicals because it is generally more conservative. The use of SR of the in vitro effect data as a second classification factor is recommended for QIVIVE_nom and QIVIVE_free models because a clearer picture can be obtained with respect to the likelihood that a biological effect will occur and that it is not caused by nonspecific cytotoxicity.