香草酸酰胺衍生物的制备及其止血活性的动力学法研究

摘要

Three novel amide derivatives were designed and prepared by the reaction of vanillic acid with 6-aminocaproic acid ( EACA) , p-aminomethylbeozoic acid ( AMBA) or tranexamic acid ( AM-CA), respectively. The structures were identified by IR, ESI-MS, 1H NMR and 13C NMR. In order to study their hemostatic activity in vitro, the plasma recalcification time ( PRT) was determined by traditional method, fluorescence kinetics method and UV kinetics method, respectively. In order to study the stability of derivatives in blood, the interaction between derivatives and human serum albu-min ( HSA) was investigated under imitated physiological condition by fluorescence spectroscopy, UV-visible absorption spectroscopy and molecular docking, respectively. The results stated clearly that derivatives could reduce PRT significantly and exhibit obvious procoagulant effect in vitro. Mo-reover, in some concentrations, the hemostatic activity of derivativeIIwas better than AMBA. The results obtained by three different methods were in correspondence. New kinetics methods not only avoided artificial error effectually but also had good repeatability. In addition, more reasonable and credible information could be obtained in the process of serum agglutination. On the other hand, the spectra experiments showed that three derivatives could all form stable complexes with HSA respective-ly. The binding types between derivatives and HSA were all van der Waals force and hydrogen bonds. The results were accordant with molecular docking. It indicates that all the derivatives can be trans-ferred and delivered by HSA to play pharmacological effects at corresponding site. Consequently, there is a strong possibility that the three vanillylamide derivatives can be applied to practice.%以香草酸为母体,分别与6-氨基己酸( EACA)、止血芳酸( AMBA)和止血环酸( AMCA)反应,合成3种未见文献报道的香草酸酰胺衍生物,并采用IR、ESI-MS、1 H NMR和13 C NMR进行结构鉴定。以传统方法、荧光动力学法和紫外动力学法测定其对血浆复钙时间( PRT)的影响。为了研究衍生物在血液中的稳定性,在模拟生理条件下,运用荧光光谱、紫外光谱及分子对接技术研究了它分别与人血清白蛋白( HSA)的结合。结果表明,3种衍生物在一定浓度范围内都可以有效缩短PRT,具有显著的止血活性。新的动力学测定方法,不仅有效避免了传统方法引入的人为误差,重现性良好,而且判断标准更加科学,并可以获得更多血浆凝结过程中的信息。3种衍生物均主要通过氢键和范德华力与HSA结合形成稳定的复合物,表明它可通过HSA在血液中传递并运输到相应部位而发生药理作用。