Inhibition of human cytochromes P450 2A6 and 2A13 by flavonoids, acetylenic thiophenes and sesquiterpene lactones from Pluchea indica and Vernonia cinerea

Abstract

The human liver cytochrome P450 (CYP) 2A6 and the respiratory CYP2A13 enzymes play role in nicotine metabolism and activation of tobacco-specific nitrosamine carcinogens. Inhibition of both enzymes could offer a strategy for smoking abstinence and decreased risks of respiratory diseases and lung cancer. In this study, activity-guided isolation identified four flavonoids 1-4 (apigenin, luteolin, chrysoeriol, quercetin) from Vernonia cinerea and Pluchea indica, four hirsutinolide-type sesquiterpene lactones 5-8 from V. cinerea, and acetylenic thiophenes 9-11 from P. indica that inhibited CYP2A6- and CYP2A13-mediated coumarin 7-hydroxylation. Flavonoids were most effective in inhibition against CYP2A6 and CYP2A13, followed by thiophenes, and hirsutinolides. Hirsutinolides and thiophenes exhibited mechanism-based inhibition and in irreversible mode against both enzymes. The inactivation kinetic K_I values of hirsutinolides against CYP2A6 and CYP2A13 were 5.32-15.4 and 0.92-8.67 µM, respectively, while those of thiophenes were 0.11-1.01 and 0.67-0.97 µM, respectively.

Keywords: CYP2A13; CYP2A6; acetylenic thiophenes; flavonoids; sesquiterpene lactones.

Figure 1.

Chemical structures of apigenin (1), luteolin (2), chrysoeriol (3), quercetin (4), 8α-(2-methylacryloyloxy)-hirsutinolide-13-O-acetate (5), 8α-tigloyloxyhirsutinolide-13-O-acetate (6), 8α-(4-hydroxymethacryloyloxy)-hirsutinolide-13-O-acetate (7), 8α-(4-hydroxytigloyloxy)-hirsutinolide-13-O-acetate (8), 2-(penta-1,3-diyn-1-yl)-5–(4-acetoxy-3-hydroxybuta-1-yn-1-yl) thiophene (9), 2-(prop-1-inyl)-5–(6-acetoxy-5-hydroxyhexa-1, 3-diinyl) thiophene (10), 2-(prop-1-inyl)-5–(5, 6-dihydroxyhexa-1,3-diinyl) thiophene (11). Chemical structure was produced using ChemDraw Professional 8.

Figure 2.

Time- and concentration-dependent inactivation and kinetics of inhibition of CYP2A6-mediated coumarin 7-hydroxylation by 2-(prop-1-inyl)-5–(5, 6-dihydroxyhexa-1,3-diinyl) thiophene 11 (A), and of CYP2A13-mediated coumarin 7-hydroxylation by 8α-(4-hydroxymethacryloyloxy)-hirsutinolide-13-O-acetate 7 (B), and 2-(prop-1-inyl)-5–(5, 6-dihydroxyhexa-1,3-diinyl) thiophene 11 (C). Data are represented as mean ± SD of triplicate experiments.