新型大豆苷元磺酸酯抗动脉粥样硬化功效的评价与机制

Daidzein has prevention effect to the abnormal proliferation of VSMCs, so it has anti-atherosclerosis efficacy also. But its defect in pharmaceutical and pharmacokinetic properties lead to weak effect, it cannot be used widely and effectively in clinical until now. So, enhancing its pharmaceutical properties and pharmacokinetics has important potentials for its clinical application by chemical modification. In this application, daidzein sulfonate derivatives were designed and synthesized by the kinetophore principle. By cellular tests derivatives with high prevention effect on VSMCs were screened and its mechanism will be explored. And then, the prevention effect and mechanism of the optimal drugs on atherosclerotic animal will be evaluated. Meanwhile, by studing their pharmaceutical properties, absorption and metabolism mechanism, in vivo pharmacokinetics, structure-activity relationship of pharmacokinetic phase of derivatives will be expored. Based on the results of activity experiments, the relationship between sulfonate, pharmacokinetic properties and efficacy will be revealed, and the novel anti-atherosclerotic candidates will be selected. Currently, the optimization of natural products were stopped on preparation of derivatives and effect at cell level, but the key factors which affect the macroscopic effects of drugs - behavior in pharmacokinetic phase - were not studied enough. The affects of sulphonate group to pharmaceutical properties and Pharmacokinetics, and to activities will be clarified in this study according to the experimental results. And, the relations between structure, behaviors in pharmacokinetic phase and activities were revealed in deeper order, to look for a new door for shortening the exploration process for new chemical entities.

大豆苷元对血管平滑肌细胞增殖有抑制作用而起到抗动脉粥样硬化的保健功效，但由于药代性质缺陷导致药效低而难以在临床上有效使用。因此，通过化学修饰改善药代性质，提高药效对其临床应用具有重要意义。本研究利用药动团变换原理设计、高选择性合成新的大豆苷元磺酸酯；利用亲和色谱技术确证大豆苷元在细胞的作用靶点与机制，筛选作用更强的大豆苷元磺酸酯进行整体动物实验评价其疗效；同时对衍生物的药学性质、药代动力学研究，揭示衍生物的药动相构效关系，进一步揭示药代性质与药效之间的关系，优选抗动脉粥样硬化的新候选药物。目前对天然产物的优化主要停留在修饰物的制备与细胞药效层面，而对影响药效的关键因素—药动相行为研究不够，本研究将着重阐释磺酸酯基的引入对衍生物的药动相行为的影响, 进而对药效的影响规律，从更深层次上揭示结构、药动相行为与活性之间的关系，尽可能的为寻找新化学实体做出探索。

大豆苷元对血管平滑肌细胞增殖有抑制作用而起到抗动脉粥样硬化的保健功效，但由于药代性质缺陷导致药效低而难以在临床上有效使用。因此，通过化学修饰改善药代性质，提高药效对其临床应用具有重要意义。本研究利用药动团变换原理设计、高选择性合成了大豆苷元脂溶性衍生物14个，确证了2个化合物的晶体结构。利用亲和色谱技术、MALDI-TOF和Sybyl X 2.0完成了大豆苷元衍生物的细胞靶蛋白的实验研究，初步清楚大豆苷元在血管平滑肌细胞的生物学靶点分子量范围。完成部分衍生物的体外抑制血管平滑肌细胞增殖与迁移作用研究，发现绝大多数大豆苷元衍生物体外抑制血管平滑肌细胞增殖活性的增加，大豆苷元4’-OH的保存更有利提高其活性。发现大豆苷元可能通过下调周期蛋白依赖性激酶，上调激酶抑制蛋白的表达来调控周期进程，从而影响细胞增殖。完成部分衍生物的体外抗炎抗氧化研究，发现大多数大豆苷元衍生物体外具有优异的抗炎活性。完成部分衍生物的抗家兔AS的研究，表明大多数衍生物具有优良的抗AS功效，机制研究表明，大豆苷元衍生物抑制VSMC增殖与迁移生物活性大幅度提高，同时经大豆苷元衍生物治疗后，主动脉局部炎症反应得到抑制，大豆苷元可能是通过抑制VSMC增殖与迁移和抗炎双重作用达到抗AS作用的。利用HPLC-MS完成了衍生物的溶解度、脂水分配系数、细胞吸收代谢，利用MALDI-TOF完成小鼠体内原位代谢研究，结合相关活性研究结果，初步揭示大豆苷元磺酸酯药动相的构效关系；进一步揭示药物的药理作用与其结构、药动相行为的内在本质联系。研究表明，大豆苷元衍生物因其磺酸酯药代团的引入，增强了其脂溶性，优化了分子结构（与靶蛋白特异性结合），更利于被细胞吸收通过细胞膜到达靶标从而使其抑制VSMC增殖迁移生物活性提高与通过JNK/AP-1通路发挥抗炎作用，从而表现出良好的抗AS作用。当然从抗炎的角度研究大豆苷元衍生物的抗AS治疗机制还需进一步研究。目前合成的14个大豆苷元衍生物均具有临床前抗AS研究价值。