Departamento de Química, Universidade Federal de São Carlos, CP 676, São Carlos, 13565-905, SP, Brazil.
J Med Chem. 2013 Mar 14;56(5):2038-44. doi: 10.1021/jm301732a. Epub 2013 Mar 4.
The use of immobilized capillary enzyme reactors (ICERs) for online ligand screening has been adopted as a new technique for high-throughput screening (HTS). In this work, the selected target was the enzyme acetylcholinesterase (AChE), and the AChE-ICERs produced were used in a liquid chromatograph-tandem ion-trap mass spectrometer. The activity and kinetic parameters were evaluated by monitoring the choline's precursor ion (M + H)(+)m/z 104.0 and its ion fragment (C2H3OH) - (M + H)(+)m/z 60.0. The assay method was validated using the reference AChE inhibitors tacrine and galanthamine. Two new ligands, out of a library of 17 coumarin derivatives, were identified, and the half-maximal inhibitory concentration (IC50), inhibition constant (K(i)), and the inhibition mechanism were determined. A coumarin derivative with IC50 similar to tacrine was highlighted.
固定化毛细管酶反应器（ICERs）用于在线配体筛选已被采用为高通量筛选（HTS）的新技术。在这项工作中，选择的靶标是酶乙酰胆碱酯酶（AChE），并在液相色谱-串联离子阱质谱仪中使用所产生的 AChE-ICERs。通过监测胆碱前体离子（M+H）（+）m/z 104.0 和其离子碎片（C2H3OH）-（M+H）（+）m/z 60.0 来评估活性和动力学参数。使用参考 AChE 抑制剂他克林和加兰他敏验证了该测定方法。从 17 种香豆素衍生物文库中鉴定出两种新的配体，并确定了半最大抑制浓度（IC50）、抑制常数（K（i））和抑制机制。具有类似于他克林的 IC50 的香豆素衍生物被突出显示。
