WY-14643, a novel antiplatelet and antithrombotic agent targeting the GPIbα receptor 

WY-14643, a novel antiplatelet and antithrombotic agent targeting the GPIbα receptor

Chen Cao a); Qingyuan Yang b); Xiaoshuang Xia a); Zhuangzhuang Chen a); Peilin Liu a); Xiaowen Wu b); Hu Hu c); Zhongren Ding b) *); Xin Li a) **)

a) Department of Neurology, The Second Hospital of Tianjin Medical University, Tianjin 300211, China

b) School of Pharmacy, Tianjin Medical University, Tianjin 300070, China

c) Department of Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310012, China

Background and purpose:

Hypolipidemia and platelet activation play key roles in atherosclerotic diseases. Pirinixic acid (WY-14643) was originally developed as a lipid-lowering drug. Here we focused on its antiplatelet and antithrombotic abilities and the underlying mechanism.

Experimental approach:

The effects of WY-14643 on platelet aggregation was measured using a lumiaggregometer. Clot retraction and spreading on fibrinogen were also assayed. PPARα􀀀 /􀀀 platelets were used to identify the target of WY-14643. The interaction between WY-14643 and glycoprotein Ibα (GPIbα) was detected using cellular thermal shift assay (CETSA), surface plasmon resonance (SPR) spectroscopy and molecular docking. GPIbα downstream signaling was examined by Western blot. The antithrombotic effect was investigated using mouse mesenteric arteriole thrombosis model. Mouse tail bleeding model was used to study its effect on bleeding side effects.

Key results:

WY-14643 concentration-dependently inhibits human washed platelet aggregation, clot retraction, and spreading. Significantly, WY-14643 inhibits thrombin-induced activation of human washed platelets with an IC50 of 7.026 μM. The antiplatelet effect of WY-14643 is mainly dependent of GPIbα. CESTA, SPR and molecular docking results indicate that WY-14643 directly interacts with GPIbα and acts as a GPIbα antagonist. WY-14643 also inhibits phosphorylation of PLCγ2, Akt, p38, and Erk1/2 induced by thrombin. Noteworthily, 20 mg/kg oral administration of WY-14643 inhibits FeCl3-induced thrombosis of mesenteric arteries in mice similarly to clopidogrel without increasing bleeding.

Conclusion and implications:

WY-14643 is not only a PPARα agonist with lipid-lowering effect, but also an antiplatelet agent as a GPIbα antagonist. It may have more significant therapeutic advantages than current antiplatelet agents for the treatment of atherosclerotic thrombosis, which have lipid-lowering effects without bleeding side effects.