title: 'Slow-binding' sixth-ligand inhibitors of cytochrome P-450 aromatase. Studies with 19-thiomethyl- and 19-azido-androstenedione.
abstract: The progress curves for the inhibition of aromatase by 19-thiomethylandrostenedione and 19-azidoandrostenedione were found to be non-linear where the extent of inhibition increased with time. Further experiments enabled these compounds to be classified as 'slow-binding' inhibitors of aromatase. The phenomenon was attributed to the formation of an initial E.I complex that rearranged to another species (E.I*) in which the interaction between the enzyme and inhibitor had been maximized, giving rise to tighter binding. When 19-thiomethylandrostenedione was used as the inhibitor the t0.5 (half-time) for the dissociation of E.I* was calculated to be 12.6 min with Ki and Ki* values of 2.4 and 1.4 nM respectively. In the case of 19-azidoandrostenedione, the two separate dissociation constants were not determined, and a single Ki value of 5 nM was obtained. The conclusions drawn from kinetic studies were confirmed by absorption spectrometry, when time-dependent formation of complexes between aromatase and either 19-thiomethylandrostenedione or 19-azidoandrostenedione were observed by the formation of 'Type II' spectra. The two complexes respectively had maxima at 429 and 418 nm. The spectral data suggested that the two inhibitors interact with the haem iron of aromatase, forming hexaco-ordinated species for which structural models are presented.
authors.affiliation: Department of Biochemistry, University of Southampton, U.K.
authors.forename: J N
authors.initials: JN
authors.lastname: Wright
authors.forename: G
authors.initials: G
authors.lastname: Slatcher
authors.forename: M
authors.initials: M
authors.lastname: Akhtar
mesh_terms: D000735:Androstenedione / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D001141:Aromatase / Q000302:isolation & purification / Q000378:metabolism*; D047072:Aromatase Inhibitors; D001665:Binding Sites; D005260:Female; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008433:Mathematics; D008861:Microsomes / Q000201:enzymology; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011485:Protein Binding
chemical_list: D047072:Aromatase Inhibitors; D008024:Ligands; C067446:19-azidoandrost-4-ene-3,17-dione; D000735:Androstenedione; C067447:19-thiomethylandrost-4-ene-3,17-dione; D001141:Aromatase