Effective Prevention of Arterial Thrombosis with Albumin-Thrombin Inhibitor Conjugates.
Effective Prevention of Arterial Thrombosis with Albumin-Thrombin Inhibitor Conjugates.
复制标题
使用白蛋白-凝血酶抑制剂结合物有效预防动脉血栓形成。
DOI：
10.1021/acs.molpharmaceut.3c00325
复制
发表时间：
2023
影响因子：
4.9
通讯作者：
Glassman,PatrickM
中科院分区：
文献类型：
--
作者：
Marcos-Contreras,OscarA;Myerson,JacobW;Nong,Jia;Brenner,JacobSamuel;Muzykantov,VladimirR;Glassman,PatrickM
血栓预防适用于血栓性疾病风险较高的患者，通常使用直接口服抗凝剂或低分子量肝素。我们假设，短暂的血栓预防（数天-数周）可以提供单剂量的抗凝剂设计延长药代动力学。在目前的工作中，d-苯丙氨酰-L-脯氨酰-L-精氨酸氯甲基酮（PPACK）被用作模型抗凝剂，以测试的假设，即凝血酶抑制剂的表面的共轭白蛋白将提供持久的保护，防止血栓性损伤。使用点击化学在白蛋白和PPACK之间形成共价缀合物，并使用凝血酶活性测定和凝块形成测定在体外对它们进行测试。在化学诱导（FeCl 3）和缺血-再灌注（短暂性大脑中动脉闭塞; tMCAO）后的动脉血栓形成小鼠模型中测试血栓预防功效。白蛋白-PPACK结合物在两种体外试验中都显示出纳摩尔效力，静脉注射后循环延长。偶联物不影响正常小鼠的止血（剪尾）或全身凝血参数。在FeCl 3诱导的血栓形成之前静脉注射缀合物提供了对大脑中动脉和颈总动脉闭塞的显著保护，并且在tMCAO模型中，在缺血-再灌注后立即注射使损伤后3天测量的每搏输出量减少约40%。本文提供的数据支持使用白蛋白相关抗凝剂作为可注射、长循环、安全的血栓预防剂。特别是，白蛋白-PPACK提供了显著的保护，防止由多种机制诱导的血栓形成，而不会对止血产生不利影响。
Thromboprophylaxis is indicated in patients at an elevated risk of developing thrombotic disorders, typically using direct oral anticoagulants or low-molecular-weight heparins. We postulated that transient thromboprophylaxis (days–weeks) could be provided by a single dose of an anticoagulant engineered for prolonged pharmacokinetics. In the present work,d-phenylalanyl-l-prolyl-l-arginine chloromethyl ketone (PPACK) was used as a model anticoagulant to test the hypothesis that conjugation of thrombin inhibitors to the surface of albumin would provide durable protection against thrombotic insults. Covalent conjugates were formed between albumin and PPACK using click chemistry, and they were testedin vitrousing a thrombin activity assay and a clot formation assay. Thromboprophylactic efficacy was tested in mouse models of arterial thrombosis, both chemically induced (FeCl3) and following ischemia-reperfusion (transient middle cerebral artery occlusion; tMCAO). Albumin-PPACK conjugates were shown to have nanomolar potency in bothin vitroassays, and following intravenous injection had prolonged circulation. Conjugates did not impact hemostasis (tail clipping) or systemic coagulation parameters in normal mice. Intravenous injection of conjugates prior to FeCl3-induced thrombosis provided significant protection against occlusion of the middle cerebral and common carotid arteries, and injection immediately following ischemia-reperfusion reduced stroke volume measured 3 days after injury by ∼40% in the tMCAO model. The data presented here provide support for the use of albumin-linked anticoagulants as an injectable, long-circulating, safe thromboprophylactic agent. In particular, albumin-PPACK provides significant protection against thrombosis induced by multiple mechanisms, without adversely affecting hemostasis.