CHARACTERIZATION OF THE PROTEIN-BINDING OF CHIRAL DRUGS BY HIGH-PERFORMANCE AFFINITY-CHROMATOGRAPHY - INTERACTIONS OF R-IBUPROFEN AND S-IBUPROFEN WITH HUMAN SERUM-ALBUMIN
CHARACTERIZATION OF THE PROTEIN-BINDING OF CHIRAL DRUGS BY HIGH-PERFORMANCE AFFINITY-CHROMATOGRAPHY - INTERACTIONS OF R-IBUPROFEN AND S-IBUPROFEN WITH HUMAN SERUM-ALBUMIN
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DOI：
10.1016/0021-9673(94)01009-4
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发表时间：
1995-02-17
影响因子：
4.1
通讯作者：
WAINER, IW
中科院分区：
文献类型：
--
作者：
HAGE, DS;NOCTOR, TAG;WAINER, IW
采用区带洗脱和高效亲和层析技术研究了右旋和右旋布洛芬与人血清白蛋白（HSA）的不同结合特性。这通过在含有已知浓度的R-或S-布洛芬作为竞争剂的移动的相存在下将少量R-和S-布洛芬注射到固定化HSA柱上来完成。这些研究表明，R-和S-布洛芬在固定化HSA柱上有一个共同的结合位点。此外，S-布洛芬至少有一个其他的主要结合区域。在pH6.9和25 ℃下，R-布洛芬与HSA的缔合平衡常数为5.3.10（5）M（-1）。在相同条件下，S-布洛芬在两个位点的缔合常数分别为1.1.10（5）M（-1）和1.2.10（5）M（-1）。S-布洛芬位点以约1：1的比例存在，并且似乎在高S-布洛芬浓度下表现出一些变构相互作用。在这项工作中使用的色谱技术是一个通用的，它可以适用于研究其他手性化合物与HSA或其他蛋白质的相互作用。
Zonal elution and high-performance affinity chromatography were used to study the different binding characteristics of R- and S-ibuprofen with the protein human serum albumin (HSA). This was done by injecting small amounts of R- and S-ibuprofen onto an immobilized HSA column in the presence of a mobile phase that contained a known concentration of R- or S-ibuprofen as a competing agent. These studies indicated that R- and S-ibuprofen had one common binding site on the-immobilized HSA column. In addition, S-ibuprofen had at least one other major binding region. The association equilibrium constant for R-ibuprofen with HSA was found to be 5.3.10(5) M(-1) at pH 6.9 and 25 degrees C. Under the same conditions, the association constants for S-ibuprofen at its two sites were 1.1.10(5) M(-1) and 1.2.10(5) M(-1). The S-ibuprofen sites were present in about a 1:1 ratio and appeared to exhibit some allosteric interactions at high S-ibuprofen concentrations. The chromatographic technique used in this work is a general one which can be adapted for use in studying the interactions of other chiral compounds with either HSA or additional proteins.