BF061, A Novel Antiplatelet And Antithrombotic Agent Targeting P2Y₁₂ And PDE

Though antiplatelet drugs are proven beneficial to coronary heart disease and stroke patients, more effective and safer antiplatelet drugs are still needed. Previously we reported that an adenine derivative BF081 inhibits platelet activation via P2Y12 antagonism and PDE inhibition. By introducing a pentose group into BF0801, we developed a novel antiplatelet agent BF061. In the range of 0.10-100μM, BF061 concentration-dependently inhibited platelet aggregation induced by 100 nM 2MeSADP. At 10μM BF061 blocked platelet aggregation and ATP release induced by 0.5 mM arachidonic acid or 1μM U46619. Compared to BF0801. BF061 is more potent in inhibiting platelet aggregation and dense granule release induced by ADP, 2MeSADP, collagen, arachidonic acid and U46619 in human washed platelets. BF061 antagonized intracellular cAMP decrease induced by 2MeSADP and further increased forskolin-elicited increase in human platelets. Basal and sodium prusside-stimulated intracellular cGMP levels in human washed platelets were also enhanced by BF061. Similar to AR-C69931MX, BF061 exhibited P2Y12 antagonism activity as evidenced by inhibiting the interaction between ADP and P2Y12 receptor expressed in CHO-K1 cells measured by atomic force microscopy. Similar to phosphodiesterase (PDE) inhibitor IBMX,BF061 robustly inhibited platelet PDE activity as measured by HPLC. Interestingly, despite being structurally similar to BF061, AR-C69931MX had no effect on human platelet PDE. Finally, we evaluated the antithrombotic role of B061 in mice using intravital microscopy. In the FeCl3 mesenteric arterial thrombosis model, a bolus injection of BF061 (25 mg/kg, intravenous) dramatically prolonged the stable thrombus formation time from 116±10 seconds to 418±55 seconds. BF061 also reduced thrombus volume in the laser-injured cremaster arteriole model. In contrast, BF061 induced dramatically less bleeding at antithrombotic dose compared to clopidogrel. In summary, we developed a novel antiplatelet and antithrombotic agent targeting both P2Y12 and PDE. Given the prevalence of combined antiplatelet therapy in clinical practice, an antiplatelet agent bearing dual activities may have therapeutic advantage as a potential antithrombotic drug.