Microsomal alcohol oxygenase catalyzes the stereoselective oxidation of 7α- and 7β-hydroxy-Δ⁸-tetrahydrocannabinol (7α- and 7β-hydroxy-Δ⁸-THC) to 7-oxo-Δ⁸-THC in monkey liver, and the activity for 7β-hydroxy-Δ⁸-THC is relatively higher than that for 7α-hydroxy-Δ⁸-THC. We previously reported that purified P450JM-E, assumed to be CYP3A8, is a major enzyme responsible for the oxidation of 7-hydroxy-Δ⁸-THC to 7-oxo-Δ⁸-THC in monkey liver and is capable of catalyzing the oxidative reaction by NADH as well as NADPH. In the present study, we demonstrated that some steroids such as testosterone and progesterone stimulated both the NADH- and NADPH-dependent conversions of 7β-hydroxy-Δ⁸-THC to 7-oxo-Δ⁸-THC in monkey liver microsomes. Kinetic analyses revealed that both the NADH- and NADPH-dependent 7-oxo-Δ⁸-THC formation showed sigmoid kinetics. Testosterone caused a decrease in S₅₀ and an increase in V_max for the NADH-dependent activity, and resulted in a decrease in S₅₀ without changing the V_max for the NADPH-dependent activity. On the other hand, NADH-dependent testosterone 6β-hydroxylation activity showed Michaelis-Menten kinetics and was also inhibited by 7β-hydroxy-Δ⁸-THC, resulting in a decrease in V_max with no effect on the K_m. NADPH-dependent testosterone 6β-hydrozylation activity was also inhibited by 7β-hydroxy-Δ⁸-THC, resulting in a decrease in both S₅₀ and V_max. In order to explain the metabolic interaction between 7β-hydroxy-Δ⁸-THC and testosterone, we propose a kinetic model involving at least three binding sites, for the mechanism of activation by testosterone.

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