Selective degradation of platelet BTK by PROTAC NX-5948 provides antithrombotic benefits without affecting haemostasis

Current antithrombotic therapies are effective in reducing thrombotic events but are limited by their associated risk of bleeding. BTK acts as a key signalling switch that drives platelet activation during thrombosis but is largely dispensable for routine haemostasis. It is an important non-redundant signalling mediator downstream of the GPVI and CLEC-2 receptors, plays a key role in thrombosis with minimal involvement in haemostasis, making it an attractive antithrombotic target. While BTK inhibitors effectively reduce thrombosis, their clinical use has been limited due to off-target effects. Protein degraders may overcome this limitation by enabling the ubiquitin proteasomal system to selectively target and degrade BTK. We here assessed the ability of the BTK degraders NX-2127 and NX-5948, currently in clinical trials for B cell pathologies, to target platelet BTK for degradation. NX-2127 and NX-5948 induced concentration-dependent degradation of BTK in washed platelets, platelet-rich plasma and whole blood. NX-5948 showed no hook effect and outperformed NX-2127 in potency, efficacy, and degradation kinetics. TMT proteomic analysis confirmed selective BTK degradation by NX-5948 with no evidence of major off-target effects. BTK degradation impaired CRP-mediated integrin IIb{beta}3 activation, P-selectin expression, platelet aggregation and in vitro thrombosis, with PAR-1 mediated platelet function being left intact. Dosing mice with NX-5948 led to efficacious degradation of platelet BTK and impaired CRP-, but not thrombin-, mediated ex vivo platelet function. In vivo, arterial thrombosis was markedly reduced, without an increase in bleeding time. Together, these results highlight NX-5948 as a potent, selective BTK degrader with antithrombotic potential and minimal haemostatic impact. Key PointsO_LIThe BTK degrader NX-5948 potently and selectively degrades platelet BTK and suppresses thrombus formation without affecting bleeding. C_LIO_LITargeting BTK degradation offers a new antithrombotic strategy that spares haemostasis and may aid patients intolerant to DAPT. C_LI