Microsomal Metabolism of Delavirdine: Evidence for Mechanism-Based Inactivation of Human Cytochrome P450 3A

• Published in: The Journal of pharmacology and experimental therapeutics Vol. 287; no. 1; pp. 381 - 388
• Main Authors: Voorman, R L, Maio, S M, Payne, N A, Zhao, Z, Koeplinger, K A, Wang, X
• Format: Journal Article
• Language: English
• Published: United States American Society for Pharmacology and Experimental Therapeutics 01.10.1998
• Subjects: AIDS/HIV; Animals; Anti-HIV Agents - metabolism; Aryl Hydrocarbon Hydroxylases; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme Inhibitors; Delavirdine - metabolism; Delavirdine - pharmacology; Electrophoresis, Polyacrylamide Gel; Humans; Kinetics; Male; Microsomes, Liver - metabolism; Oxidoreductases, N-Demethylating - antagonists & inhibitors; Rats; Rats, Sprague-Dawley
• ISSN: 0022-3565; 1521-0103

Administration of delavirdine, an HIV-1 reverse transcriptase inhibitor, to rats or monkeys resulted in apparent loss of hepatic microsomal CYP3A and delavirdine desalkylation activity. Human CYP3A catalyzes the formation of desalkyl delavirdine and 6′-hydroxy delavirdine, an unstable metabolite, while CYP2D6 catalyzes only desalkyl delavirdine. CYP2D6 catalyzed desalkyl delavirdine formation was linear with time (up to 30 min) but when catalyzed by cDNA expressed CYP3A4 or human liver microsomes the reaction rate declined progressively with time. Coincubation with triazolam showed that delavirdine caused a time- and NADPH-dependent loss of CYP3A4 activity in human liver microsomes as measured by triazolam 1′-hydroxylation. The catalytic activity loss was saturable and was characterized by a K i of 21.6 ± 8.9 μM and a k inact of 0.59 ± 0.08 min − 1 . An apparent partition ratio of 41 was determined with cDNA expressed CYP3A4, based on the substrate depletion method. Incubation of [ 14 C]delavirdine with microsomes from several species resulted in irreversible association with an approximately 50 kDa protein, as demonstrated by SDS-PAGE/autoradiography. Binding to the protein was NADPH dependent, glutathione insensitive, proportional to the level of CYP3A expression and was inhibited by ketoconazole, a specific CYP3A inhibitor. NADPH-dependent irreversible binding to human and rat total microsomal protein was demonstrated following exhaustive extraction of microsomal protein. Binding was decreased in the presence of glutathione and appeared to be related to expression level of CYP3A. These results suggest that delavirdine can inactivate CYP3A and has the potential to slow the metabolism of coadministered CYP3A substrates.