RT Journal Article
SR Electronic
T1 Proluciferin Acetals as Bioluminogenic Substrates for Cytochrome P450 Activity and Probes for CYP3A Inhibition
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 2403
OP 2410
DO 10.1124/dmd.111.041541
VO 39
IS 12
A1 Meisenheimer, Poncho L.
A1 Uyeda, H. Tetsuo
A1 Ma, Dongping
A1 Sobol, Mary
A1 McDougall, Mark G.
A1 Corona, Cesear
A1 Simpson, Dan
A1 Klaubert, Dieter H.
A1 Cali, James J.
YR 2011
UL http://dmd.aspetjournals.org/content/39/12/2403.abstract
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.