Synthesis and characterization of a photoaffinity labelling probe based on the structure of the cystic fibrosis drug ivacaftor
Synthesis and characterization of a photoaffinity labelling probe based on the structure of the cystic fibrosis drug ivacaftor
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DOI：
10.1016/j.tet.2018.06.016
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发表时间：
2018-09-20
期刊：
影响因子：
2.1
通讯作者：
Young, Robert N.
中科院分区：
文献类型：
--
作者：
Hamilton, C. Michael;Hung, Maurita;Young, Robert N.
囊性纤维化（CF）是由编码囊性纤维化跨膜传导调节因子（CFTR）蛋白的基因的功能丧失突变引起的遗传性疾病。依伐卡托（1）是第一个批准用于治疗CF的治疗药物，能够恢复某些CFTR变体的门控活性，尽管对其作用机制知之甚少。在本文中，我们描述了基于并入光反应性二氮杂环丙烷部分的依伐卡托的结构的光亲和标记（PAL）探针（2）的合成，用于设计用于鉴定突变CFTR上依伐卡托的结合位点的标记研究。PAL探针2保留了增强活性，其效力与1相似，使用荧光成像板读取器（FLIPR（R））测定测量野生型（Wt）-CFTR的离子电导增强。以人血清白蛋白（HSA）为模型蛋白的光标记实验表明，探针2可以以与观察到的和预测的结合一致的方式标记HSA。(C)2018爱思唯尔有限公司版权所有
Cystic Fibrosis (CF) is a genetic disorder caused by loss-of-function mutations to the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Ivacaftor (1) was the first therapeutic approved for the treatment of CF that is able to restore gating activity to certain CFTR variants although the mechanism of action is poorly understood. Herein we describe the synthesis of a photoaffinity labelling (PAL) probe (2) based on the structure of ivacaftor incorporating a photoreactive diazirine moiety for use in labelling studies designed to identify the binding site for ivacaftor on mutant CFTR. The PAL probe 2 retained potentiation activity, with a potency similar to 1, using a Fluorescent Imaging Plate Reader (FLIPR (R)) assay measuring ion conductance potentiation of wild type (Wt)-CFTR. Photolabelling experiments with human serum albumin (HSA) as a model protein have shown that probe 2 can label HSA in a manner consistent with observed and predicted binding. (C) 2018 Elsevier Ltd. All rights reserved.