Hemostasis-Independent targeted treatment of thrombosis

 描述(申请人提供)：血栓性疾病，包括心肌梗死(MI)、中风和肺栓塞(PE)，是全球主要的死亡原因。目前的抗血小板、抗凝和纤溶药物在减少缺血并发症方面是有效的，但与严重出血相关，从而对死亡率产生不利影响。我们建议的理由是，定向抗血栓药物输送将增强疗效和 减少出血。最近，通过与血小板受体CD36的结合，MRP-14被确定为血栓形成的直接调节因子。因此，MRP-14靶向作为一种双重治疗方法具有潜力-即，它可以通过阻断MRP-14与CD36的结合来直接阻断血栓形成，以及用于纳米颗粒导向的溶栓剂输送。重要的是，与纤维蛋白等替代靶点不同，MRP-14缺乏对多种止血参数没有影响，例如尾部出血时间、凝血酶生成或血小板黏附/扩散。这项建议使用组合方法来确定同时与MRP-14结合的多肽，同时阻止与CD36的结合。我们将使用自然启发的烟草花叶病毒(TMV)，这是一种300×18 nm的杆状植物病毒纳米颗粒，我们已经证明由于形状效应，它具有增强血栓归巢的作用。中心假设是MRP-14靶向将允许抑制血栓形成和准确地递送 携带药物的纳米颗粒，从而减少溶栓和止血所需的有效剂量。这一假说将通过具有以下两个目标的光化学小鼠血栓形成模型来验证：将确定针对MRP-14的特异性多肽以阻断其下游效应(目标1)，以及将利用最有前途的多肽来靶向血栓形成，并将在体外和体内评估治疗效果(目标2)。这种方法是创新的，因为它将是第一个开发出MRP-14靶向多肽的公司，这种多肽能够同时干扰血小板中的MRP-14：CD36信号，并靶向血栓的溶栓剂。该项目为临床翻译铺平了道路，利用已经获得FDA批准的阿尔替普酶，并通过积极提供来提高其安全性和有效性。这些目标的完成将导致靶向载体系统的产生，该系统将解决当前的临床挑战，b为出血有限的血栓形成提供更有效的治疗方法。

 DESCRIPTION (provided by applicant): Thrombotic disorders, including myocardial infraction (MI), stroke, and pulmonary embolism (PE), are the leading causes of death worldwide. Current antiplatelet, anticoagulant, and fibrinolytic agents are effective at reducing ischemic complications but are associated with significant bleeding that adversely impacts mortality. The rationale for our proposal is that targeted anti-thrombotic drug delivery will enhance efficacy and reduce bleeding. Recently, MRP-14 was identified as a direct regulator of thrombosis through its association with the platelet receptor CD36. Therefore, MRP-14 targeting has potential as a dual-therapeutic approach - namely, it can be used to directly block thrombosis by interrupting MRP-14 binding to CD36 as well as for nanoparticle-directed delivery of a thrombolytic agent. Importantly, unlike alternate targets such as fibrin, MRP-14 deficiency has no effect on multiple hemostatic parameters, such as tail bleeding time, thrombin generation, or platelet adhesion/spreading. This proposal uses a combinatorial approach to determine peptides that simultaneously bind to MRP-14 while blocking binding to CD36. We will use nature-inspired tobacco mosaic virus (TMV), a 300 by 18 nm rod-shaped plant virus-based nanoparticle we have shown to have enhanced thrombus homing due to shape-based effects. The central hypothesis is that MRP-14 targeting will allow for inhibition of thrombosis and precise delivery of drug-bearing nanoparticles, thereby reducing the effective dose necessary for thrombolysis and limiting bleeding. This hypothesis will be tested with a photochemical mouse thrombosis model with the following two aims: peptides specific for MRP- 14 that blocks its downstream effects will be identified (Aim 1), and the the most promising peptides will be utilized to target thrombosis and the therapeutic efficacy will be evaluated in vitro and in vivo (Aim 2). This approach is innovative, as it will be the first to develop an MRP-14 targeting peptide that is capable of simultaneously disrupting MRP-14:CD36 signaling in platelets and targeting a thrombolytic agent to thrombus. This project paves the way for clinical translation, utilizing already FDA-approved alteplase and improving its safety and efficacy through active delivery. Completion of these aims will result in a targeted carrier system that will address current clinical challenges b providing a more efficacious therapeutic approach for thrombosis with limited bleeding.