FG-8mer抑制内源性因子X酶的分子机制及其机制相关的抗凝活性特征

岩藻糖化糖胺聚糖(FG)是化学结构特殊的天然产物，具有强效内源性因子X酶(f.Xase)抑制活性。由于内源性f.Xase是新的低出血倾向抗血栓药物的靶点，FG化学及药理学研究具有重要的学术价值和应用价值。持续近40年的FG研究成果丰硕，但其确切化学结构和抑制f.Xase活性的分子机制是仍尚待逾越的关键性学术难题。本课题前期在阐明FG确切结构研究中取得重要突破，发现了目前所知的化学结构最简单的选择性f.Xase抑制剂FG-8mer。在此条件下，本课题拟采用酶促动力学、靶蛋白相互作用、荧光及FRET法蛋白结构分析以及靶蛋白共结晶制备等方法，希望通过全面系统的分析研究，清晰揭示FG-8mer强效抑制f.Xase的分子机制，解决FG药理学关键学术问题，为f.Xase抑制剂的发现提供新的策略思路；并进一步研究FG-8mer分子机制相关的抗凝抗血栓活性特点，分析和评价f.Xase抑制剂的潜在应用价值。

Fucosylated glycosaminoglycan (FG) is a natural product with unique chemical structure, which can potently inhibit tenase (factor Xase, f.Xase) of the intrinsic coagulation pathway. The intrinsic f.Xase has been identified as a novel target for antithrombotic agents with low bleeding tendency, therefore, chemical and pharmacological study of FG is important both for academic value and potential application. During the past 40 years, although academic research on FG has achieved great progress, the precise chemical structure and molecular mechanism of f.Xase inhibition are the conundrums that remains to be solved. Our previous work has made a significant breakthrough in illuminating the precise structure of FG, and discovered that FG-8mer is the selective f.Xase inhibitor with the simplest chemical structure so far known. Based on these, this project is intended to study more comprehensively and systemically to reveal the molecular mechanism of f.Xase inhibition by FG-8mer, using the technical methods, such as enzymic kinetics, protein-compound interaction, fluorescence method and FRET for protein structural analysis, and analysis of co-crystal f.IXa and FG-8mer. Additionally, we will further explore the anticoagulant and antithrombotic characteristic of FG-8mer associated with the molecular mechanism, as well as evaluate the potential application value of f.Xase inhibitor. This study will provide useful information and strategy for the discovery of novel f.Xase inhibitors.