Chromatographic studies of drug interactions with alpha(1)-acid glycoprotein by ultrafast affinity extraction and peak profiling.
Chromatographic studies of drug interactions with alpha(1)-acid glycoprotein by ultrafast affinity extraction and peak profiling.
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DOI：
10.1016/j.chroma.2017.03.056
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发表时间：
2017-05-12
期刊：
影响因子：
--
通讯作者：
Hage DS
中科院分区：
文献类型：
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作者：
Beeram S;Bi C;Zheng X;Hage DS
与血清蛋白如α1-酸性糖蛋白(AGP)的相互作用可对药物的行为和药代动力学产生重大影响。用AGP微柱进行超快亲和萃取和峰形分析，考察了几种模型药物(氯丙嗪、异丙吡胺、丙咪嗪、利多卡因、心得安和维拉帕米)的这一过程。在pH7.4和37℃条件下，用超快亲和萃取法测定药物与可溶性琼脂糖蛋白的结合平衡常数为10~4-106M−1，与文献值符合较好。其中一些值取决于使用单位点结合模型时存在的药物和蛋白质的相对浓度；这些结果表明实际上存在更复杂的混合模式相互作用，这也被用于分析数据。采用单位点模型时，用超快亲和萃取法测得的表观解离速率常数为0.2 4~7.0 S−1，并与药物和蛋白质的相对浓度有关。当药物与蛋白质的相对量减少时，该方法获得了较低的表观解离速率常数，其结果与低药物浓度下的峰分布测得的结果接近。这些信息应该有助于更好地了解这些药物和其他药物在循环中如何与AGP相互作用。此外，在本报告中优化和使用的用于检查这些系统的层析方法也可以用于其他具有生物医学意义的溶质-蛋白质相互作用的分析。
Interactions with serum proteins such as alpha1-acid glycoprotein (AGP) can have a significant effect on the behavior and pharmacokinetics of drugs. Ultrafast affinity extraction and peak profiling were used with AGP microcolumns to examine these processes for several model drugs (i.e., chlorpromazine, disopyramide, imipramine, lidocaine, propranolol and verapamil). The association equilibrium constants measured for these drugs with soluble AGP by ultrafast affinity extraction were in the general range of 104–106 M−1 at pH 7.4 and 37°C and gave good agreement with literature values. Some of these values were dependent on the relative drug and protein concentrations that were present when using a single-site binding model; these results suggested a more complex mixed-mode interaction was actually present, which was also then used to analyze the data. The apparent dissociation rate constants that were obtained by ultrafast affinity extraction when using a single-site model varied from 0.24 to 7.0 s−1 and were dependent on the relative drug and protein concentrations. Lower apparent dissociation rate constants were obtained by this approach as the relative amount of drug versus protein was decreased, with the results approaching those measured by peak profiling at low drug concentrations. This information should be useful in better understanding how these and other drugs interact with AGP in the circulation. In addition, the chromatographic approaches that were optimized and used in this report to examine these systems can be adapted for the analysis of other solute-protein interactions of biomedical interest.