基于血浆蛋白结合率硫酸氢氯吡格雷片与血塞通分散片协同作用

Synergistic Effects of Clopidogrel and Xuesaitong Dispersible Tablet by Modulating Plasma Protein Binding

目的:以抗血小板聚集效应物质人参皂苷Rb1为对象,研究硫酸氢氯吡格雷片对血塞通分散片血浆蛋白结合率影响。方法:首先建立PBS透析外液、健康受试者空白血清中人参皂苷Rb1的HPLC测量方法,选用平衡透析考查血塞通单用和联合氯吡格雷血浆蛋白结合率的差异;以家兔血清白蛋白(PDB ID 3V09)为模板选用同源模建方法构建受试者空白血清白蛋白(HSA)三维空间结构,用PRO-CHECK和ERRAT方法对两种蛋白合理性进行评价,采用分子模拟技术研究上述药物与HSA结合情况,多层面考查血塞通及氯吡格雷在血浆蛋白结合率方面的作用特征。结果:血塞通在高(人参皂苷Rb1含量5.0μg/m L)、中(人参皂苷Rb1含量1.0μg/m L)、低(人参皂苷Rb1含量0.4μg/m L)下与健康受试者空白血清蛋白结合率分别为(58.17±3.82)%、(57.43±3.21)%、(55.63±3.42)%,当合并加入含2.0μg/m L硫酸氢氯吡格雷的片剂后血塞通血浆蛋白结合率分别下降为(46.54±3.35)%、(49.25±3.56)%、(48.15±3.76)%,分子模拟结果显示化合物均与健康受试者空白血清白蛋白存在不同程度的竞争结合效应。结论:氯吡格雷对血塞通血浆蛋白结合率存在影响。

Objective： The ginsenoside Rb1,which account for platelet aggregation of Xuesaitong dispersible tablet,was selected to investigate the synergistic effects of clopidogrel（ CPG） and Xuesaitong dispersible tablet drug by modulating plasma protein binding rate aspect. Methods： The HPLC and equilibrium dialysis were employed to determine the concentration of Rb1 both in dialysate（ PBS） and blank plasma from healthy volunteer blood donors. The differences in protein-binding rate between Xuesaitong dispersible tablet alone（ the concentration of ginsenoside Rb1 were 5. 0,1. 0,0. 4 μg / m L,respectively） and combined with CPG（ each add CPG 2 μg / m L） were then compared. The three-dimensional spatial structure of the blank plasma albumin（ HSA） in the subjects was construct by rabbit plasma albumin（ PDB ID 3V09） template and evaluated by PRO-CHECK and ERRAT methods. Molecular simulation technique was used to display the competition mechanism with human plasma protein. Results： The protein binding rate of Xuesaitong dispersible tablet alone group in plasma PBS and human plasma at high（ the concentration of ginsenoside Rb1 were 5. 0 μg / m L）,middle（ the concentred of ginsenoside Rb1 were 1. 0 μg / m L） and low（ the concentration of ginsenoside Rb1 were 0. 4 μg / m L） concentrations were（ 58. 17 ±3. 82） %,（ 57. 43 ± 3. 21） %,（ 55. 63 ± 3. 42） % respectively. When combined with CPG（ each add CPG 2 μg / m L）,the protein binding rate value were decline to（ 46. 54 ± 3. 35） %,（ 49. 25 ± 3. 56） %,（ 48. 15 ± 3. 76） %,respectively. The molecular simulation results suggested that the two compounds have competitive synergistic effects with human plasma protein. Conclusion： The present investigation suggestes that there are synergistic effects of CPG and Xuesaitong dispersible tablet by modulating plasma protein binding rate of ginsenoside Rb1.