Inhibition and inactivation of equine aromatase by steroidal and non-steroidal compounds. A comparison with human aromatase inhibition.
In order to approach the detailed structure-function relationships of aromatase, we studied the inhibitory and inactivatory potencies of several steroidal androstenedione analogues (1: 4-hydroxyandrostenedione, 2: 4-acetoxyandrostenedione and 3: 7 alpha-(4'-amino)phenylthio-4-androstene-3, 17-dione) and non-steroidal imidazole derivatives (4: ketoconazole, 5: miconazole and 6: fadrozole) on equine aromatase in placental microsomes, a well established mammalian model. Human placental microsomes and the purified enzyme from equine testis were also used to compare inhibition by 1 and 2. In equine microsomes, all compounds tested exhibited a competitive inhibition, with Ki values of 4.1, 26 and 1.8 nM for 1, 2 and 3, and of 2400, 1.4 and 4 nM for 4, 5, and 6, respectively. The Km for androstenedione, the substrate mainly used in these studies, was 1.8 +/- 0.13 nM. The three non-steroidal derivatives did not inactivate equine aromatase, but 1 and 2 acted as comparable inactivators to a much higher degree than 3. Compound 1 inhibited in a similar manner (89-94%) purified or equine and human microsomal aromatases, whereas 2 inhibited microsomal aromatase more efficiently in the horse than in man (92% and 33% inhibition, respectively). There was only a 40% inhibition with 2 on the purified equine enzyme, which is no more in the natural membrane environment. The comparisons between equine and human microsomal aromatases allow precise functional and structural differences to be observed with these enzymes.
Moslemi
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Laboratoire de Biochimie et Biologie Moléculaire, EP CNRS 9, IBBA, Université de Caen, France.
Seralini
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D000735:Androstenedione / Q000031:analogs & derivatives / Q000494:pharmacology*; D000818:Animals; D047072:Aromatase Inhibitors; D004789:Enzyme Activation / Q000187:drug effects; D017316:Fadrozole / Q000494:pharmacology; D006736:Horses; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D007654:Ketoconazole / Q000494:pharmacology; D008297:Male; D008825:Miconazole / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects
D047072:Aromatase Inhibitors; D007093:Imidazoles; D000735:Androstenedione; C028626:7-(4'-amino)phenylthioandrostenedione; D008825:Miconazole; D017316:Fadrozole; C014594:formestane; D007654:Ketoconazole