Adenosine Receptor Agonist HE-NECA Enhances Antithrombotic Activities of Cangrelor and Prasugrel in vivo by Decreasing of Fibrinogen Density in Thrombus

Abstract

Blood platelets’ adenosine receptors (AR) are considered to be a new target for the anti-platelet therapy. This idea is based on in vitro studies which show that signaling mediated by these receptors leads to a decreased platelet response to activating stimuli. In vivo evidence for the antithrombotic activity of AR agonists published to date were limited, however, to the usage of relatively high doses given in bolus. The present study was aimed at verifying if these substances used in lower doses in combination with inhibitors of P2Y12 could serve as components of dual anti-platelet therapy. We have found that a selective A2A agonist 2-hexynyl-5’-N-ethylcarboxamidoadenosine (HE-NECA) improved the anti-thrombotic properties of either cangrelor or prasugrel in the model of ferric chloride-induced experimental thrombosis in mice. Importantly, HE-NECA was effective not only when applied in bolus as other AR agonists in the up-to-date published studies, but also when given chronically. In vitro thrombus formation under flow conditions revealed that HE-NECA enhanced the ability of P2Y12 inhibitors to decrease fibrinogen content in thrombi, possibly resulting in their lower stability. Adenosine receptor agonists possess a certain hypotensive effect and an ability to increase the blood–brain barrier permeability. Therefore, the effects of anti-thrombotic doses of HE-NECA on blood pressure and the blood–brain barrier permeability in mice were tested. HE-NECA applied in bolus caused a significant hypotension in mice, but the effect was much lower when the substance was given in doses corresponding to that obtained by chronic administration. At the same time, no significant effect of HE-NECA was observed on the blood–brain barrier. We conclude that chronic administration of the A2A agonist can be considered a potential component of a dual antithrombotic therapy. However, due to the hypotensive effect of the substances, dosage and administration must be elaborated to minimize the side-effects. The total number of animals used in the experiments was 146.

Potential for modulation of platelet function via adenosine receptors during inflammation

“…These studies were performed using non‐selective or selective adenosine A 2A/2B receptor agonists (2Cl‐Ado, CV‐1808, NECA, CGS 21680, HE‐NECA, regadenoson, MRE 0094, UK 432097, PSB 0777 and LUF 5835), most of which are commercially available, in combination with the P2Y 12 receptor antagonists (cangrelor, the prasugrel metabolite R‐138727, and ticagrelor). The vast majority of the examined adenosine analogues was found to possess antiplatelet activity and/or have the ability to improve the action of P2Y 12 antagonists (cangrelor and prasugrel metabolite) in preventing platelet activation, aggregation or thrombus formation (Boncler et al, 2019; Polak et al, 2021; Wolska et al, 2019). At the intracellular level, stimulation of adenosine A 2A receptor, in addition to blockade of P2Y 12 receptor, resulted in further elevation of intracellular cAMP and inhibition of calcium mobilization caused by P2Y 12 inhibitors (Wolska et al, 2020).…”