Zhu Junfeng, Yi Xiaojiao, Huang Peng, Chen Shuqing, Wu Yongjiang
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, PR China.
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, PR China.
J Pharm Biomed Anal. 2017 Feb 5;134:100-107. doi: 10.1016/j.jpba.2016.11.028. Epub 2016 Nov 18.
Danhong injection (DHI) is a widely used Chinese medicine injection (CMI) for the clinical treatment of cardiovascular and cerebrovascular diseases. In this study, a simple and efficient in vitro method based on ultrafiltration LC-MS and molecular modeling has been developed to study the human serum albumin (HSA) binding of the compounds in DHI. Seven major components including protocatechuic aldehyde, p-coumaric acid, salvianolic acid D, rosmarinic acid, salvianolic acid E, lithospermic acid and salvianolic acid B were identified as HSA ligands and their binding degrees in the proposed non-saturated model were 26.17, 37.69, 99.77, 91.78, 96.91, 99.42 and 98.10%, respectively. Considering the drug-HSA binding property of the compounds in DHI may change during drug combination therapy, competitive binding assay was carried out to evaluate the influence of aspirin on the DHI-HSA binding. Experimental results revealed that the salvianolic acids in DHI had stronger binding ability to HSA than sodium salicylate. To further verify the results above, molecular modeling and probe displacement assay were conducted to investigate the optimum binding site and binding affinity of the ligands on HSA. Our findings suggested that the established method could be a powerful tool to study the drug-HSA binding property of CMIs.
丹红注射液（DHI）是一种广泛用于临床治疗心脑血管疾病的中药注射剂。在本研究中，已开发出一种基于超滤液相色谱 - 质谱联用和分子模拟的简单高效体外方法，用于研究丹红注射液中化合物与人血清白蛋白（HSA）的结合情况。原儿茶醛、对香豆酸、丹酚酸D、迷迭香酸、丹酚酸E、紫草酸和丹酚酸B这七种主要成分被鉴定为HSA配体，它们在提出的非饱和模型中的结合度分别为26.17%、37.69%、99.77%、91.78%、96.91%、99.42%和98.10%。考虑到丹红注射液中化合物的药物 - HSA结合特性在联合药物治疗过程中可能会发生变化，进行了竞争性结合试验以评估阿司匹林对丹红注射液 - HSA结合的影响。实验结果表明，丹红注射液中的丹酚酸对HSA的结合能力比水杨酸钠更强。为了进一步验证上述结果，进行了分子模拟和探针置换试验，以研究配体在HSA上的最佳结合位点和结合亲和力。我们的研究结果表明，所建立的方法可能是研究中药注射剂药物 - HSA结合特性的有力工具。
