Adenosine-mediated inhibition of platelet aggregation by acadesine. A novel antithrombotic mechanism in vitro and in vivo.
Inhibition of platelet aggregation by acadesine was evaluated both in vitro and ex vivo in human whole blood using impedance aggregometry, as well as in vivo in a canine model of platelet-dependent cyclic coronary flow reductions. In vitro, incubation of acadesine in whole blood inhibited ADP-induced platelet aggregation by 50% at 240 +/- 60 microM. Inhibition of platelet aggregation was time dependent and was prevented by the adenosine kinase inhibitor, 5'-deoxy 5-iodotubercidin, which blocked conversion of acadesine to its 5'-monophosphate, ZMP, and by adenosine deaminase. Acadesine elevated platelet cAMP in whole blood, which was also prevented by adenosine deaminase. In contrast, acadesine had no effect on ADP-induced platelet aggregation or platelet cAMP levels in platelet-rich plasma, but inhibition of aggregation was restored when isolated erythrocytes were incubated with acadesine before reconstitution with platelet-rich plasma. Acadesine (100 mg/kg i.v.) administered to human subjects also inhibited platelet aggregation ex vivo in whole blood. In the canine Folts model of platelet thrombosis, acadesine (0.5 mg/kg per min, i.v.) abolished coronary flow reductions, and this activity was prevented by pretreatment with the adenosine receptor antagonist, 8-sulphophenyltheophylline. These results demonstrate that acadesine exhibits antiplatelet activity in vitro, ex vivo, and in vivo through an adenosine-dependent mechanism. Moreover, the in vitro studies indicate that inhibition of platelet aggregation requires the presence of erythrocytes and metabolism of acadesine to acadesine monophosphate (ZMP).
Bullough
D A
DA
Department of Cardiovascular Pharmacology, Gensia Inc., San Diego, California 92121.
Zhang
C
C
Montag
A
A
Mullane
K M
KM
Young
M A
MA
D000241:Adenosine / Q000502:physiology*; D000243:Adenosine Deaminase / Q000494:pharmacology; D000248:Adenosine Kinase / Q000037:antagonists & inhibitors; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001241:Aspirin / Q000494:pharmacology; D001790:Blood Physiological Phenomena; D003328:Coronary Thrombosis / Q000097:blood; D003331:Coronary Vessels / Q000187:drug effects / Q000502:physiology; D004176:Dipyridamole / Q000494:pharmacology; D004195:Disease Models, Animal; D004285:Dogs; D004912:Erythrocytes / Q000187:drug effects; D006801:Humans; D008297:Male; D010949:Plasma / Q000502:physiology; D010974:Platelet Aggregation / Q000187:drug effects*; D058915:Purinergic P1 Receptor Antagonists; D012039:Regional Blood Flow / Q000187:drug effects; D012263:Ribonucleosides / Q000378:metabolism / Q000494:pharmacology*; D013806:Theophylline / Q000031:analogs & derivatives / Q000494:pharmacology; D014372:Tubercidin / Q000031:analogs & derivatives / Q000494:pharmacology
C040541:5'-deoxy-5-iodotubercidin; D058915:Purinergic P1 Receptor Antagonists; D012263:Ribonucleosides; D000620:Aminoimidazole Carboxamide; C011651:acadesine; D004176:Dipyridamole; C031433:8-(4-sulfophenyl)theophylline; D013806:Theophylline; D000248:Adenosine Kinase; D000243:Adenosine Deaminase; D000241:Adenosine; D014372:Tubercidin; D001241:Aspirin