title: Aryl acetylenes as mechanism-based inhibitors of cytochrome P450-dependent monooxygenase enzymes.
abstract: Aryl acetylenes have been investigated as inhibitors of cytochrome P450 (P450)-dependent alkoxyresorufin dealkylation dealkylation activities in liver microsomes prepared from rats exposed to beta-naphthoflavone, isosafrole, or phenobarbital. Many of the acetylenes investigated produce pseudo-first-order time-dependent and NADPH-dependent losses of the dealkylation activities characteristic of mechanism-based irreversible inactivation (suicide inhibition). Replacing the terminal hydrogen of aryl acetylenes with a methyl group to convert ethynes into propynes enhances the inhibition of P450 1A enzymes; in some instances, this modification converts a reversible inhibitor of P450s into a suicide inhibitor. In contrast, ethynes are more effective suicide inhibitors of P450 2B-dependent dealkylations than the corresponding propynes. Aryl acetylenes with an ethynyl group on the 2 position of naphthalene or on the 9 position of phenanthrene and arylalkyl acetylenes with alkyl chains containing 2, 3, or 4 methylene groups are selective inhibitors of P450 2B1/2B2 in liver microsomes from rats. Aryl acetylenes also act as suicide inhibitors of P450 1A2 in human liver microsomes, of purified P450 1A2 from rabbit or rat liver in reconstituted systems, and of purified recombinant human P450 1A2 and 1A1 in reconstituted systems. 4-(1-Propynyl)biphenyl (4PBi) inactivated P450 1A2-dependent ethoxyresourfin deethylation (EROD) activity in human liver microsomes in an NADPH-dependent process (k(inactivation), 0.23 min-1; KI, 2.3 microM). 4PBi also inactivated purified recombinant human P450 1A2 (k(inactivation), 0.24 min-1; KI, 4.3 microM). In agreement with previous reports [Yun, C.-H., Hammons, G. J., Jones, G., Martin, M. V., Hopkins, N. E., Alworth, W. L., and Guengerich, F. P. (1992) Biochemistry 31, 10556-10563], 2-ethynylnaphthalene (2EN) was not a suicide inhibitor of the P450 1A2 activity in human liver microsomes but did inactivate purified human P450 1A2. Neither 4PBi nor 2EN affected diagnostic activities of human microsomal P450 2E1, 2C9/10, 3A4, or 2C19. In the systems examined, the losses of P450-dependent activity produced by these aryl acetylenes were not accompanied by corresponding decreases in the measured P450 absorption spectra. Thus P450 inactivation by these aryl acetylenes does not involve labeling and destruction of the heme. Incubation of 4PBi with microsomal P450 1A1 or 1A2 from rat liver under conditions that lead to P450-dependent, enzyme inactivations generates a 2-biphenylylpropionic acid product. This suggests that the suicide inhibition of P450s by propynylaryl acetylenes proceeds via a methylaryl ketene formed by a 1,2-methyl rearrangement, analogous to the mechanism of suicide inhibition by ethynyl acetylenes that proceed via ketene intermediates formed by 1,2-hydrogen shifts [Ortiz de Montellano, P. R., and Kunze, K. L. (1981) Arch. Biochem. Biophys. 209, 710-712].
authors.affiliation: Department of Chemistry, Tulane University, New Orleans, Louisiana 70118-5645, USA.
authors.forename: M
authors.initials: M
authors.lastname: Foroozesh
authors.forename: G
authors.initials: G
authors.lastname: Primrose
authors.forename: Z
authors.initials: Z
authors.lastname: Guo
authors.forename: L C
authors.initials: LC
authors.lastname: Bell
authors.forename: W L
authors.initials: WL
authors.lastname: Alworth
authors.forename: F P
authors.initials: FP
authors.lastname: Guengerich
mesh_terms: D000114:Acetylene / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001555:Benzene Derivatives / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000494:pharmacology; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors; D065609:Cytochrome P-450 CYP1A2 Inhibitors; D019362:Cytochrome P-450 CYP2B1 / Q000037:antagonists & inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology; D009281:Naphthalenes / Q000494:pharmacology; D010616:Phenanthrenes / Q000494:pharmacology; D011721:Pyrenes / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D013250:Steroid Hydroxylases / Q000037:antagonists & inhibitors; D013329:Structure-Activity Relationship
chemical_list: D001555:Benzene Derivatives; D001713:Biphenyl Compounds; D065609:Cytochrome P-450 CYP1A2 Inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D009281:Naphthalenes; D010616:Phenanthrenes; D011721:Pyrenes; D011994:Recombinant Proteins; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D019363:Cytochrome P-450 CYP1A1; D019362:Cytochrome P-450 CYP2B1; C025619:steroid 16-beta-hydroxylase; D000114:Acetylene