Inhibition of 15-lipoxygenases by flavonoids: structure-activity relations and mode of action

We have recently reported that flavonoids of cocoa inhibit the mammalian 15-lipoxygenase-1-a catalyst of enzymatic lipid peroxidation. To elucidate the structure-activity relationship of the inhibitory effect, we investigated the effects of 18 selected flavonoids of variable structure on pure rabbit reticulocyte and soybean 15-lipoxygenases using linoleic acid as substrate. Moreover, the inhibition by quercetin was studied in detail to gain insight into the mode of action. Quercetin was found to modulate the time-course of the reaction of both lipoxygenases by three distinct effects: (i) prolongation of the lag period, (ii) rapid decrease in the initial rate after the lag phase was overcome, (iii) time-dependent inactivation of the enzyme during reaction but not in the absence of substrate. A comparison of the IC(50) for the rapid inhibition of rabbit reticulocyte 15-lipoxygenase-1 revealed that (i) the presence of a hydroxyl group in the flavonoid molecule is not essential, (ii) a catechol arrangement reinforces the inhibitory effect, (iii) in the presence of a catechol arrangement the inhibitory potency inversely correlates with the number of hydroxyl groups, (iv) a 2,3-double bond in the C ring strengthens the inhibitory effect. The flavone luteolin turned out to be the most potent inhibitor of the mammalian enzyme with an IC(50) of 0.6 microM followed by baicalein (1 microM) and fisetin (1.5 microM).