PT  - JOURNAL ARTICLE
AU  - Poncho L. Meisenheimer
AU  - H. Tetsuo Uyeda
AU  - Dongping Ma
AU  - Mary Sobol
AU  - Mark G. McDougall
AU  - Cesear Corona
AU  - Dan Simpson
AU  - Dieter H. Klaubert
AU  - James J. Cali
TI  - Proluciferin Acetals as Bioluminogenic Substrates for Cytochrome P450 Activity and Probes for CYP3A Inhibition
AID  - 10.1124/dmd.111.041541
DP  - 2011 Dec 01
TA  - Drug Metabolism and Disposition
PG  - 2403--2410
VI  - 39
IP  - 12
4099  - http://dmd.aspetjournals.org/content/39/12/2403.short
4100  - http://dmd.aspetjournals.org/content/39/12/2403.full
SO  - Drug Metab Dispos2011 Dec 01; 39
AB  - Cytochrome P450 (P450) assays use probe substrates to interrogate the influence of new chemical entities toward P450 enzymes. We report the synthesis and study of a family of bioluminogenic luciferin acetal substrates that are oxidized by P450 enzymes to form luciferase substrates. The luciferin acetals were screened against a panel of purified P450 enzymes. In particular, one proluciferin acetal has demonstrated sensitive and selective CYP3A4-catalyzed oxidation to a luciferin ester—Km and kcat are 2.88 μM and 5.87 pmol metabolite · min−1 · pmol enzyme−1, respectively. The proluciferin acetal was used as a probe substrate to measure IC50 values of known inhibitors against recombinant CYP3A4 or human liver microsomes. IC50 values for the known inhibitors correlate strongly with IC50 values calculated from the traditional high-performance liquid chromatography-based probe substrate testosterone. Luciferin acetals are rapidly oxidized to unstable hemi-orthoesters by CYP3A resulting in luciferin esters and, therefore, are conducive to simple rapid CYP3A bioluminescent assays.