8-pCPT-conjugated cyclic AMP analogs exert thromboxane receptor antagonistic properties

Carsten Sand; Maria Grandoch; Christof Börgermann; Paschal A. Oude Weernink; Yvonne Mahlke; Benjamin Schwindenhammer; Artur-Aron Weber; Jens W. Fischer; Karl H. Jakobs; Martina Schmidt

doi:10.1160/TH09-06-0341

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2010; 103(03): 662-676
DOI: 10.1160/TH09-06-0341

Cardiovascular Biology and Cell Signalling

Schattauer GmbH

8-pCPT-conjugated cyclic AMP analogs exert thromboxane receptor antagonistic properties

Carsten Sand^*

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Maria Grandoch^*

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Christof Börgermann

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Paschal A. Oude Weernink

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Yvonne Mahlke

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Benjamin Schwindenhammer

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Artur-Aron Weber

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Jens W. Fischer

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Karl H. Jakobs

¹Institut für Pharmakologie, Universitätsklinikum Essen, Essen, Germany

,

Martina Schmidt

²Department of Molecular Pharmacology, University of Groningen, Groningen, The Netherlands

› Author Affiliations

Abstract

Summary

Membrane-permeable 8-(4-chlorophenylthio)-2'-O-methyl cyclic AMP (8-pCPT-2'-O-Me-cAMP) has been shown to specifically activate cAMP-regulated Epac proteins, without direct effects on protein kinase A and protein kinase G. During isometric tension measurements in thoracic aortic rings from Wistar rats, we observed that 8-pCPT-2'-O-Me-cAMP selectively induced a rightward shift of the concentration response curve for the thromboxane mimetic U46619, without altering the contractile response to noradrenaline. We hypothesised that 8-pCPT-2'-O-Me-cAMP and similar compounds may function as direct thromboxane receptor antagonists. Indeed, in addition to 8-pCPT-2'-OMe-cAMP, also 8-pCPT-cAMP, 8-(4-chlorophenylthio)-adenosine-3',5'-cyclic monophosphorothioate, Rp-isomer (Rp-8-CPT-cAMPS) and 8-CPT-adenosine, but not 8-Bromo-2'-O-Me-cAMP, induced rightward shifts of the contractile response to U46619. Likewise, 8-pCPT-2'-O- Me-cAMP and Rp-8-CPT-cAMPS, but not 8-Bromo-2'-O-Me-cAMP, specifically reduced U46619-induced aggregation of human platelets. In addition, 8-pCPT-2'-O-Me-cAMP and Rp-8-CPT-cAMPS completely reversed U46619-induced reduction of intercellular adhesion molecule-1 expression and migration of human coronary artery endothelial cells. Most important, the cAMP analogs that reduced the contractile response to U46619 also concentration-dependently inhibited binding of the thromboxane receptor radioligand [5,6-³H]SQ29548 to human platelets. We conclude that 8-pCPT-conjugated cAMP analogs exert competitive thromboxane receptor antagonistic properties.

Keywords

Exchange protein directly activated by cyclic AMP - vascular tissue - human platelets - endothelial cells - thromboxane A₂

Full Text

References

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