Studies on the mechanism of action of the omeprazole-derived cyclic sulphenamide.
The inhibitory effects of omeprazole and omeprazole-derived metabolites were studied on Escherichia coli glutaminase activity at pH 2.5 which might represent the conditions present at the target enzyme (K+/H+-ATPase) in the secretory membrane of the intact parietal cell. Omeprazole and the omeprazole-derived cyclic sulphenamide inhibited glutaminase at pH 2.5 with identical potency (IC50 36 microM). The substrate, glutamine as well as the mercaptane, dithiothreitol, protect the enzyme. Furthermore, dithioerythritol was found to reverse inhibition. This indicates that an SH-group localized in the substrate binding center of glutaminase is most likely involved in the reaction leading to enzyme inhibition. Glutaminase inhibition by both compounds was less pronounced at pH 5.0. Omeprazole radical, the metabolite generated from the cyclic sulphenamide at more neutral pH values, failed to affect the enzyme. These findings were in contrast with the properties of the omeprazole-derived cyclic sulphenamide and radical at the K+/H+-ATPase preparation. This enzyme was inhibited by both compounds at pH 7.5 with a high potency, and reversal experiments with dithiothreitol demonstrate that these agents interfere with SH-groups of the K+/H+-ATPase. From these data it is suggested that the cyclic sulphenamide and the radical interfere by different reaction pathways with enzymatic SH-groups.
Beil
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Abteilung Allgemeine Pharmakologie, Medizinische Hochschule Hannover, Federal Republic of Germany.
Staar
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Sewing
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KF
D000251:Adenosine Triphosphatases / Q000378:metabolism; D004226:Dithioerythritol / Q000494:pharmacology; D004229:Dithiothreitol / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005609:Free Radicals; D005972:Glutaminase / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D017506:H(+)-K(+)-Exchanging ATPase; D006863:Hydrogen-Ion Concentration; D009853:Omeprazole / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D013997:Time Factors
D005609:Free Radicals; D005973:Glutamine; C055084:omeprazole cyclic sulfenamide; D004226:Dithioerythritol; D005972:Glutaminase; D000251:Adenosine Triphosphatases; D017506:H(+)-K(+)-Exchanging ATPase; D009853:Omeprazole; D004229:Dithiothreitol