Wurtzel Jeremy G T, Gray Brian D, Pak Koon Y, Zhao Xuefei, Ma Peisong, McKenzie Steven E, Tanujaya Michelle, Rizzo Victor, Del Carpio-Cano Fabiola, Rao A Koneti, Lee-Gau Chong Parkson, Goldfinger Lawrence E
Cardeza Foundation for Hematologic Research, Department of Medicine, Division of Hematology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
Molecular Targeting Technologies Inc, West Chester, Pennsylvania, USA.
J Thromb Haemost. 2025 Jan;23(1):108-122. doi: 10.1016/j.jtha.2024.10.007. Epub 2024 Oct 17.
Phosphatidylserine (PS) is a procoagulant phospholipid enriched on surfaces of activated vascular cells including platelets, endothelium, monocytes, and microvesicles. As a molecular driver of thrombosis accessible to drug blockade, PS is an attractive pharmacologic target for modulating thrombogenesis, with potentially reduced bleeding risk compared to anticoagulant and antiplatelet therapies.
Test antithrombotic capabilities of a liposomal formulation, Zn-dipicolylamine cyanine-3[22,22]/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (molar ratio, 3:97), designated as DPAL, which we previously described binds selectively to PS-enriched cell surfaces, compared with effects on bleeding, in mouse models.
PS-dependent DPAL binding to human and murine platelets was tested in vitro. Thrombosis and bleeding after DPAL intravenous administration were tested in C57Bl/6J mice following FeCl carotid arterial injury and tail tip amputation, respectively. Incorporation in hemostatic clots was investigated in the cremaster muscle laser injury model. Toxicity was tested by direct exposure to human endothelial cell cultures.
DPAL bound agonist-stimulated, PS-positive human and murine platelets, blocked by Annexin V or Ano6 deletion, which ablate PS exposure. DPAL prolonged prothrombin time, but did not prevent thrombin-induced fibrinogen receptor activation or aggregation, nor alter blood cell counts including platelets. Following arteriolar laser injury, DPAL bound wound surfaces and edges without destabilizing plugs. DPAL dose-dependently blocked FeCl-induced arterial thrombosis but did not substantially increase bleeding, or induce endothelial cell death.
DPAL reduces thrombogenesis with minimal effects on bleeding in mouse models via selective binding to PS. DPAL may support novel approaches to modulate pathogenic thrombin generation with improved safety profiles in multiple contexts.
磷脂酰丝氨酸（PS）是一种促凝磷脂，在包括血小板、内皮细胞、单核细胞和微囊泡在内的活化血管细胞表面富集。作为一种可通过药物阻断的血栓形成分子驱动因素，PS是调节血栓形成的一个有吸引力的药理学靶点，与抗凝和抗血小板治疗相比，其出血风险可能降低。
在小鼠模型中，测试一种脂质体制剂Zn-二吡啶甲胺花青素-3[22,22]/1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱（摩尔比，3:97），命名为DPAL的抗血栓形成能力，我们之前描述过它能选择性结合富含PS的细胞表面，并比较其对出血的影响。
在体外测试PS依赖性DPAL与人及小鼠血小板的结合。分别在C57Bl/6J小鼠中，通过FeCl颈动脉损伤和尾尖截肢后，测试静脉注射DPAL后的血栓形成和出血情况。在提睾肌激光损伤模型中研究其在止血凝块中的掺入情况。通过直接暴露于人内皮细胞培养物中来测试毒性。
DPAL结合激动剂刺激的、PS阳性的人及小鼠血小板，Annexin V或Ano6缺失可阻断这种结合，Annexin V或Ano6缺失会消除PS暴露。DPAL延长凝血酶原时间，但不阻止凝血酶诱导的纤维蛋白原受体激活或聚集，也不改变包括血小板在内的血细胞计数。在小动脉激光损伤后，DPAL结合伤口表面和边缘而不破坏血栓。DPAL剂量依赖性地阻断FeCl诱导的动脉血栓形成，但不会显著增加出血或诱导内皮细胞死亡。
DPAL通过选择性结合PS在小鼠模型中减少血栓形成，对出血影响最小。DPAL可能支持在多种情况下调节致病性凝血酶生成的新方法，具有更好的安全性。
