控制“总血栓风险”化合物HY023016对血小板糖蛋白受体GPIbα及凝血酶exosite II双靶点作用的抗血栓分子机制

Reduce "total thrombotic risk" of patients is expected to become the new ideas of the treating thrombotic diseases. Based on the idea,the target compound HY023016 is a novel molecular which consists of direct thrombin inhibitor dabigatran etexilate reactive groups and the effective ingredients of the traditional Chinese medicine "Chung Chong"(Ligusticum wallichii Franchat) -Tetramethylpyrazine( Chinese patent application number: 201110172073.5). Early pharmacodynamic study has found that compound HY023016 owns a extensive antithrombotic effect. We speculate that the HY023016 compound plays a synergistic antithrombotic effect, by acting on the essential to platelet activation and the burst of coagulation cascade-platelet glycoprotein receptor GPIbα and thrombin anion binding sites exosite II.On one hand, GPIbα inhibits the platelet activation mediated by itself and thrombin explosive proliferation; On the other hand as exosite II, and further inhibition of thrombin-induced platelet activation and thrombin procoagulant function, resulting in its synergistic multi-targetantithrombotic effect.The project around these two targets, the utilization of recombinant molecular techniques, flow cytometry experiments, explore the HY023016 the multi-target synergy. Through this project, will provide a scientific basis for the treating thrombotic diseases by reducing "total thrombotic risk".

降低患者"总血栓风险"有望成为血栓性疾病治疗的新思路。依据该思路，我们将直接凝血酶抑制剂达比加群与川芎嗪的活性代谢物川芎醇偶联设计合成了化合物HY023016，拥有自主知识产权。先期的药效学研究发现HY023016具有较全面的抗血栓作用。我们推测这种作用是通过作用于血小板激活与凝血瀑布爆发中至关重要的血小板糖蛋白受体GPIbα以及凝血酶阴离子结合位点exosite II得以实现。它作用于GPIbα，可抑制其介导的血小板激活和凝血酶爆发式增殖；作用于exosite II，可进一步抑制凝血酶诱导的血小板激活以及凝血酶促凝血功能，从而产生多靶点协同抗血栓作用，达到降低 "总血栓风险"的目的。本项目围绕这两个靶点，采用重组分子技术、流式细胞检测等实验方法，阐明HY023016多靶点抗血栓作用的分子机理。通过本项目的研究，将为血栓性疾病治疗新思路-降低"总血栓风险"提供科学的依据。

降低患者"总血栓风险"有望成为血栓性疾病治疗的新思路。我们利用直接凝血酶抑制剂达比加群与川芎嗪的活性代谢物川芎醇偶联设计合成了化合物HY023016，并研究血小板糖蛋白受体GPIbα与凝血酶阴离子结合位点exosite II在血栓发生发展过程中的重要地位，同时阐明化合物HY023016多靶点协同抗血栓的分子机制。通过本项目的研究，我们发现化合物HY023016通过多种途径发挥抗血栓作用：它能特异性结合人体血小板膜受体GPIbα，抑制GPIbα和vWF-A1结构域的结合、抑制凝血酶与GPIbα的结合（IC50=1.93×10-5M）；该化合物表现出较达比加群酯对凝血酶更强的亲和力，抑制凝血酶的exosite I（IC50=2.45×10-6M）和凝血酶阴离子结合位点exosite II作用较好；HY023016能够在一定程度上抑制凝血酶和肝素的结合、抑制凝血酶与纤维蛋白原γ’肽的结合（IC50=0.77±1.29 μM）、抑制凝血酶诱导的血小板激活（IC50=10.68±1.29μM）。目前，针对血小板糖蛋白受体GPIbα和凝血酶exosite II双靶点抑制药物尚未面世，化合物HY023016是我们在该领域的有益尝试。作为血小板糖蛋白受体GPIbα和凝血酶exosite II双靶点抑制剂，化合物HY023016在体内外研究中均表现出良好的抗血栓活性，我们将以此为基础对HY023016及其系列化合物的成药性进行深入研究，期望开发出以血小板糖蛋白受体GPIbα和凝血酶exosite II为靶点的有效抗血栓药物，为心血管疾病患者的血栓防治提供新的治疗方案。