Influence of vehicle, distant topical delivery, and biotransformation on the chemopreventive activity of apigenin, a plant flavonoid, in mouse skin

Purpose. This study was designed to (a) establish a short-term in vivo system to evaluate topical formulations of apigenin, (b) determine whether apigenin should be topically delivered to the local skin tissue or systemic circulation, (c) investigate if biotransformation was involved in apigenin's chemopreventive activity. Methods. The effect of topical applied apigenin in acetone/DMSO (A/D, 9:1) on the promotion of skin tumorigenesis was studied. The influence of apigenin in DMSO, A/D (4:1), and propylene glycol/DMSO (PG/D, 4:1) on 12-O-tetradecanoylphorbol-13-acetate (TPA) induced ornithine decarboxylase (ODC) activity was compared. Distant topical delivery of apigenin was conducted on abdominal skin and ODC activity was monitored in dorsal skin. Potential glucuronidation/sulfation of apigenin in intact skin was assessed by measuring isolated apigenin before and after enzyme hydrolysis with glucuronidase/sulfatase. The epidermal extracts from apigenin-treated SENCAR mice were analyzed for metabolites by HPLC. Results. Apigenin (5 mu mol) in A/D did not significantly reduce skin tumor incidence in contrast to previous data with DMSO. Inhibition of TPA-induced ODC by apigenin in three vehicles was in the order of DMSO > A/D > PG/D. TPA-induced ODC in dorsal skin was not inhibited by apigenin delivered from abdominal skin. The quantity of apigenin recovered from epidermal extract was not different before and after beta-glucuronidase/sulfatase treatment. Metabolites were not observed in the HPLC profiles of epidermal extracts from apigenin-treated mice. Conclusions. (a) The short-term TPA-induced ODC was validated for evaluating topical formulations of apigenin. (b) Topical delivery of apigenin should target the local skin tissue, (c) Glucuronidation/sulfation appeared not to be involved in apigenin's chemopreventive activity.