Studies of phenytoin binding to human serum albumin by high-performance affinity chromatography
Studies of phenytoin binding to human serum albumin by high-performance affinity chromatography
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DOI：
10.1016/j.jchromb.2004.06.012
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发表时间：
2004-09-25
影响因子：
3
通讯作者：
Hage, DS
中科院分区：
文献类型：
--
作者：
Chen, JZ;Ohnmacht, C;Hage, DS
采用高效亲和色谱法研究了苯妥英钠与固定化人血清白蛋白（HSA）的结合。这是通过前沿分析和竞争性结合区带洗脱实验完成的，后者使用了四种探针化合物，用于注射到含有已知浓度苯妥英的移动的相存在下的HSA的主要和次要结合位点。结果发现，苯妥英可以与HSA在华法林-阿扎丙宗，吲哚-苯二氮卓类，他莫昔芬，洋地黄毒苷网站的这种蛋白质。在pH 7.4和37 ℃下，苯妥英在吲哚-苯二氮卓和洋地黄毒苷位点的缔合常数分别为1.04（+/-0.05）x 10（4）M-1和6.5（+/-0.6）x 10（3）M-1。苯妥英的变构相互作用和直接结合似乎发生在华法林-阿扎丙酮和他莫昔芬位点。这种相当复杂的结合系统表明，确定特定药物的HSA上的结合区域作为了解这些物质在血液中的运输和表征其药物相互作用潜力的手段的重要性。(C)2004 Elsevier B. V.保留所有权利。
High-performance affinity chromatography was used to study the binding of phenytoin to an immobilized human serum albumin (HSA) column. This was accomplished through frontal analysis and competitive binding zonal elution experiments, the latter of which used four probe compounds for the major and minor binding sites of HSA injected into the presence of mobile phases containing known concentrations of phenytoin. It was found that phenytoin can interact with HSA at the warfarin-azapropazone, indole-benzodiazepine, tamoxifen, and digitoxin sites of this protein. The association constants for phenytoin at the indole-benzodiazepine and digitoxin sites were determined to be 1.04 (+/-0.05) x 10(4) M-1 and 6.5 (+/-0.6) x 10(3) M-1, respectively, at pH 7.4 and 37 degreesC. Both allosteric interactions and direct binding for phenytoin appear to take place at the warfarin-azapropazone and tamoxifen sites. This rather complex binding system indicates the importance of identifying the binding regions on HSA for specific drugs as a means for understanding the transport of such substances in blood and in characterizing their potential for drug-drug interactions. (C) 2004 Elsevier B.V. All rights reserved.