Potently Inhibit Ultimate Carcinogen Formation from Benzo[ St. John's Wort Extracts and Some of Their Constituents

Commercially available St. John’s wort (Hypericum perforatum) preparations and some of their main constituents (hypericin, pseudohypericin, hyperforin, rutin, and quercetin) were examined for their potential to inhibit carcinogen activation by human cytochrome P450 1A1 (CYP1A1). We used a reconstituted system consisting of purified human CYP1A1, purified human NADPH-cytochrome P450 reductase, and dilaurylphosphatidylcholine as lipid component. St. John’s wort extracts potently inhibited CYP1A1-catalyzed ( )-trans-7,8-dihydro-7,8-dihydroxy-benzo(a)pyrene (7,8-diol-B[a]P) epoxidation, the terminal reaction leading to the ultimate carcinogenic product ( )-B[a]P-r-7,t-8-dihydrodiol-t-9,10epoxide (diolepoxide 2). All constituents, except rutin, were shown to possess strong inhibitory potencies toward diolepoxide 2 formation from 7,8-diol-B[a]P, with IC50 values of 0.5 M (hypericin), 1.2 M (hyperforin), 1.5 M (quercetin), and 8 M (pseudohypericin), respectively. Preincubation experiments revealed that their action was not mechanism based. Inhibition kinetics studies showed the anthrodianthrone compound hypericin to be a noncompetitive inhibitor, with a Ki value of 0.6 M, and the phloroglucinol hyperforin to be a competitive inhibitor, with a Ki value of 1.1 M. When the effects on NADPH-P450 reductase activity were investigated, all constituents of St. John’s wort studied turned out to be rather ineffective inhibitors; quercetin was the only exception, with an IC50 value of 20 M. These in vitro data indicate that St. John’s wort extracts and some of their constituents potently inhibit the major human procarcinogen-activating enzyme CYP1A1.