Studies on aromatase inhibition with 4-androstene-3,6,17-trione: Its 3β-reduction and time-dependent irreversible binding to aromatase with human placental microsomes

• Published in: Journal of steroid biochemistry Vol. 28; no. 3; pp. 337 - 344
• Main Authors: Numazawa, Mitsuteru, Tsuji, Masachika, Mutsumi, Ayako
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.09.1987
• Subjects: Androstenes - metabolism; Androstenes - pharmacology; Aromatase Inhibitors; Female; Humans; Kinetics; Microsomes - enzymology; Oxidation-Reduction; Placenta - enzymology; Protein Binding; Time Factors
• ISSN: 0022-4731
• DOI: 10.1016/0022-4731(87)91028-4

The metabolism of 4-androstene-3,6,17-trione (AT), previously described as a suicide substrate for aromatase, and its irreversible binding to aromatase were studied by using human placental microsomes. AT was rapidly converted into 3β-reduced metabolite (3-OHAT) with an enzyme other than aromatase in the microsomes in the presence of NADPH under either aerobic or anaerobic conditions. The conversion was efficiently prevented by a steroid 5α-reductase inhibitor. 3-OHAT was characterized as a competitive ( K i = 6.5 μM) and irreversible inhibitor of aromatase. Both 14C-labeled AT and 3-OHAT were demonstrated to be irreversibly bound to aromatase probably through a sulfur atom of the enzyme in time-dependent manners in the presence of NADPH, being accompanied with time-dependent losses of the enzyme activity. It was shown that the process of an apparent time-dependent loss of aromatase activity caused by AT even under conditions allowing its 3β-reduction should principally depend on the action of the parent inhibitor AT itself and not on that of the metabolite 3-OHAT.