Sowell J, Mason J C, Strekowski L, Patonay G
Department of Chemistry, Georgia State University, Atlanta, USA.
Electrophoresis. 2001 Aug;22(12):2512-7. doi: 10.1002/1522-2683(200107)22:12<2512::AID-ELPS2512>3.0.CO;2-9.
Drug binding to serum albumin influences several important pharmacological properties such as toxicity, solubility, activity, distribution, and excretion. It is therefore of interest to have methodologies that allow for the determination of drug-albumin affinity constants while simultaneously providing information on the location of the drug binding site. In the present work we describe a method for the determination of binding constants of drugs known to bind to subdomain IIIA of serum albumin. Drugs used in the study were ketoprofen, ibuprofen, quinidine, naproxen, imipramine, and clofibrate. Binding constants of the drugs were determined by near-infrared dye-displacement capillary electrophoresis. The dye-displacement technique uses a competitive-type interaction between the drug of interest and a dye probe to arrive at a binding constant. A heptamethine cyanine dye was used as a probe for drug binding at subdomain IIIA of serum albumin. The utility of the dye as a noncovalent label for serum albumin was investigated. Additionally, the ability of the method to illustrate enantioselective binding is shown. The dye displacement technique has advantages over current electrophoresis-based techniques in that it is faster and uses less reagent.
药物与血清白蛋白的结合会影响多种重要的药理学特性，如毒性、溶解度、活性、分布和排泄。因此，拥有能够测定药物 - 白蛋白亲和常数并同时提供药物结合位点位置信息的方法很有意义。在本研究中，我们描述了一种用于测定已知与血清白蛋白亚结构域IIIA结合的药物结合常数的方法。研究中使用的药物有酮洛芬、布洛芬、奎尼丁、萘普生、丙咪嗪和氯贝丁酯。药物的结合常数通过近红外染料置换毛细管电泳法测定。染料置换技术利用目标药物与染料探针之间的竞争型相互作用来得出结合常数。一种七甲川花菁染料被用作血清白蛋白亚结构域IIIA上药物结合的探针。研究了该染料作为血清白蛋白非共价标记的效用。此外，还展示了该方法说明对映体选择性结合的能力。染料置换技术相对于当前基于电泳的技术具有优势，因为它速度更快且使用的试剂更少。
