Comparative Assessment of the Inhibition of Recombinant Human CYP19 (Aromatase) by Azoles Used in Agriculture and as Drugs for Humans

Abstract

Azoles (imidazoles and triazoles) are used as antifungal agents in agriculture and in medicine, and also for antiestrogen therapy, e.g., for breast cancer treatment. Antifungal activity is based on inhibition of fungal CYP51 (lanosterol 14α‐demethylase), and estrogen biosynthesis reduction is due to azole inhibition of CYP19 (aromatase). Inhibition of aromatase by antifungal agents is usually an unwanted side effect and may cause endocrine disruption. A fluorimetric assay based on human recombinant CYP19 enzyme with dibenzylfluorescein as a substrate was used to compare the inhibitory potency of 22 azole compounds. Dose responses were established and duplicate datasets were analyzed with a nonlinear mixed‐effects model with cumulative normal distribution for the logarithm of concentration. IC50 values (50% inhibitory concentration) of 13 fungicides used in agriculture ranged more than 700‐fold, starting from 0.047 µM. The potency of seven human drugs spanned more than 7000‐fold, starting from 0.019 µM. Most potent fungicides included prochloraz, flusilazole, and imazalil, and most potent medicinal antifungals were bifonazole, miconazole, and clotrimazole. These in vitro data indicate that the top‐ranking azoles used as antifungal agents or drugs are as potent inhibitors of aromatase as are antiestrogen therapeutics used to treat breast cancer. These putative effects of azole agents and drugs on steroid biosynthesis and sex hormone balance should be considered when used in human subjects and also in wildlife exposed to azole fungicides used in agriculture.

• Azoles
• Antifungals
• Pesticides
• CYP19
• Aromatase
• Monooxygenase
• Steroid metabolism
• Estrogen biosynthesis
• Endocrine disruption
• Enzyme inhibition

Acknowledgments

This work was supported by the Swiss Federal Office of Public Health (grants no. 02.000451 and 02.001534)