A novel triple combination of pharmacological chaperones improves F508del-CFTR correction
A novel triple combination of pharmacological chaperones improves F508del-CFTR correction
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DOI：
10.1038/s41598-018-29276-y
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发表时间：
2018-07-30
影响因子：
4.6
通讯作者：
Thomas, David Y.
中科院分区：
文献类型：
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作者：
Carlile, Graeme W.;Yang, Qi;Thomas, David Y.
直接结合F508 del-CFTR并纠正其错误定位的药理学伴侣（例如VX-809，lumacaftor）是囊性纤维化（CF）的有希望的治疗剂。然而，迄今为止，单个校正剂仅提供了以FEV 1测量的肺功能的4%的改善，这表明需要多种药物来实现实质性的临床益处。在这里，我们检查是否存在多个药理学伴侣位点，并且可以靶向增强F508 del-CFTR的拯救，前提是与单一校正剂相比，多个药理学伴侣的相加或协同拯救表明它们具有不同的作用位点。首先，我们发现药理学伴侣VX-809和RDR 1的组合提供了F508 del-CFTR的相加校正。然后使用细胞热稳定性测定（CETSA），我们使用新型药理学伴侣工具化合物4-甲基-N-[3-（吗啉-4-基）喹喔啉-2-基]苯磺酰胺（MCG 1516 A）证明了第三个重要位点的可能性。所有三种药理学伴侣似乎与第一个核苷酸结合结构域（NBD 1）相互作用。MCG 1516 A、RDR 1和VX-809的三重组合使CFTR功能恢复至良好分化的HBE细胞中非CF细胞的> 20%，并且在其他细胞类型中恢复至高得多的水平。因此，结果表明在F508 del-CFTR NBD 1上存在至少三个不同的药理学伴侣位点，这鼓励了三重校正剂组合的开发。
Pharmacological chaperones (e.g. VX-809, lumacaftor) that bind directly to F508del-CFTR and correct its mislocalization are promising therapeutics for Cystic Fibrosis (CF). However to date, individual correctors provide only similar to 4% improvement in lung function measured as FEV1, suggesting that multiple drugs will be needed to achieve substantial clinical benefit. Here we examine if multiple sites for pharmacological chaperones exist and can be targeted to enhance the rescue of F508del-CFTR with the premise that additive or synergistic rescue by multiple pharmacological chaperones compared to single correctors indicates that they have different sites of action. First, we found that a combination of the pharmacological chaperones VX-809 and RDR1 provide additive correction of F508del-CFTR. Then using cellular thermal stability assays (CETSA) we demonstrated the possibility of a third pharmacologically important site using the novel pharmacological chaperone tool compound 4-methyl-N-[3-(morpholin-4-yl)quinoxalin-2-yl] benzenesulfonamide (MCG1516A). All three pharmacological chaperones appear to interact with the first nucleotide-binding domain (NBD1). The triple combination of MCG1516A, RDR1, and VX-809 restored CFTR function to > 20% that of non-CF cells in well differentiated HBE cells and to much higher levels in other cell types. Thus the results suggest the presence of at least three distinct sites for pharmacological chaperones on F508del-CFTR NBD1, encouraging the development of triple corrector combinations.