Microsomal prostaglandin E2 synthase-1 (MPGES1) catalyzes the formation of prostaglandin E2 from the endoperoxide prostaglandin H2. MPGES1 expression is induced in inflammatory diseases, and this enzyme is regarded as a potential drug target. To aid in the drug discovery effort, a simple method for determination of inhibition mechanism and potency toward both prostaglandin H2 and glutathione (GSH) has been developed. Using an assay with thiobarbituric acid-based detection, the inhibitory effects of six MPGES1 inhibitors were evaluated. The IC50 values obtained at three substrate (S) concentrations ([S]<KM, [S]≈KM, [S]>KM) were used to estimate inhibition modality and inhibition constant values. This facilitated strategy is a useful and general screening method to evaluate the inhibitory effects of new drug compounds.
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