PB0761 - Differential interactions of energy metabolism inhibitors – CORM-A1 and 3PO – with cangrelor in terms of platelet antiaggregatory effect

Background: A major strategy to reduce cardiovascular events related to thrombosis is antiplatelet therapy, but some patients with metabolic diseases show platelet hyperreactivity despite treatment. The hypothesis of our study is that the pharmacological regulation of platelet energy metabolism augments the action of antiplatelet drugs, which do not affect energy metabolism per se. Such an approach can help reduce their effective doses and unwanted side effects.

Aims: The aim was to investigate the effects of partial inhibition of platelet energy metabolism on the antiaggregatory action of cangrelor.

Methods: Washed platelets, isolated from blood donated by healthy volunteers, were exposed to carbon monoxide releasing molecule A1 (CORM-A1; inhibits platelet aggregation by inhibiting mitochondrial respiration and glycolysis) or 3PO (an inhibitor of the glycolytic enzyme 6-PhosphoFructo-2-Kinase/Fructose-2,6-Biaphosphatase 3) and cangrelor (an inhibitor of the purinergic P2Y12 receptor), individually and in combination. Platelet aggregation was measured by light transmission aggregometry on a multiwall plate. Mitochondrial respiration and glycolysis rates were measured using the Seahorse XF technique, while intraplatelet metabolites and eicosanoids levels were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Results: Individually added CORM-A1, 3PO and cangrelor inhibited platelet aggregation with EC20: 84.6 ± 8.90 µM, 12.2 ± 1.56 µM and 0.48 ± 0.127 nM, respectively. Cangrelor, in contrast to CORM-A1 and 3PO, did not affect energy metabolism pathways in platelets. CORM-A1 activated the pentose phosphate pathway and reduced the production of thromboxane A2 and other eicosanoids, while 3PO induced the production of prostaglandins D2, E2, and F2α, without affecting thromboxane A2. The combined treatment of platelets with cangrelor and, respectively, CORM-A1 or 3PO, showed distinct effects: CORM-A1 synergistically enhanced the antiaggregatory effect of cangrelor, while the effect of 3PO was only additive.

Conclusion(s): We showed that pharmacological attenuation of platelet energy metabolism resulted in a more potent antiplatelet effect of cangrelor.

Funding
The National Science Centre Poland [2021/41/B/NZ7/01426]