Direct inhibition of cyclooxygenase-1 and -2 by the kinase inhibitors SB 203580 and PD 98059 -: SB 203580 also inhibits thromboxane synthase

The kinase inhibitors SE 203580 and PD 98059 have been reported to be specific inhibitors of the 38- and 42/44-kDa mitogen-activated protein kinase (MAPK) pathways, respectively. In this study, the two inhibitors were found to decrease platelet aggregation induced by low concentrations of arachidonic acid, suggesting that they also interfere with the metabolism of arachidonic acid to thromboxane A(2). In support of this, SE 203580 and PD 98059 inhibited the conversion of exogenous [H-3]arachidonic acid to [H-3]thromboxane in intact platelets. Measurement of platelet cyclooxygenase-1 activity following immunoprecipitation revealed that SE 203580 and PD 98059 are direct inhibitors of this enzyme. Both compounds were shown to inhibit purified cyclooxygenase-1 and -2 by a reversible mechanism, In addition, SE 203580 (but not PD 98059) inhibited platelet aggregation induced by prostaglandin H-2 and the conversion of prostaglandin H-2 to thromboxane A(2) in intact platelets. SE 203580 also inhibited this pathway in platelet microsome preparations, suggesting a direct inhibitory effect on thromboxane synthase. These results demonstrate that direct effects of the two kinase inhibitors on active arachidonic acid metabolites have to be excluded before using these compounds for the investigation of MAPKs in signal transduction pathways. This is of particular relevance to studies on the regulation of cytosolic phospholipase A(2) as these two MAPKs are capable of phosphorylating cytosolic phospholipase A(2), thereby increasing its intrinsic activity.