Lunaccio Dario, Allegretta Caterina, Pesce Emanuela, Scarano Naomi, Vinci Virginia, Salis Annalisa, Tasso Bruno, Brullo Chiara, Capitanio Nazzareno, Piccoli Claudia, Pedemonte Nicoletta, Cichero Elena, Millo Enrico, Laselva Onofrio
Department of Experimental Medicine, Section of Biochemistry, School of Medical and Pharmaceutical Sciences, University of Genoa, Genoa, Italy.
Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
Biochem Pharmacol. 2025 Jul 9;240:117127. doi: 10.1016/j.bcp.2025.117127.
Although remarkable rescue has been achieved for treatment of Cystic Fibrosis (CF) by the combination of two correctors (VX-661, VX-445) and one potentiator (VX-770), the stability and trafficking defects induced by the most common mutation, F508del, are not completely reversed. Therefore, more effective CFTR correctors are still needed. We employed in silico and molecular modelling approaches to design and probe the binding site of novel series of CFTR correctors (a-c). Structure-based studies allowed us to design and synthesize novel class I (b series) and class II (a series) modulators. Thus, class I modulator activity relies on interactions with Met152, Phe81, Phe191, Trp361. The design of class II corrector could be managed via NBD2-ligand H-bonds, involving Gln1291 or Val1288. Furthermore, c compounds were proposed featuring putative dual corrector ability (2c) and class II corrector behavior (1c). Functional measurements in F508del-CFTR CFBE cells and primary nasal epithelial cells demonstrated that eight of fourteen compounds acted as CFTR correctors and the F508del-CFTR rescue was comparable to the level measured after VX-809 or VX-445 treatment in CFBE cells. Through rational selection based on molecular docking studies and mechanisms of action, we showed that combination of compounds (7a+1b and 2a+2b) targeting distinct domains of CFTR, can additively/synergistically rescue F508del-CFTR function in both CFBE cell line and primary nasal cells. Our study demonstrated that in silico and in vitro approaches to develop and investigate the mechanism of action of novel CFTR correctors could be a tool to optimize the combination correctors therapy to synergistically rescue mutated CFTR.
尽管通过两种校正剂（VX-661、VX-445）和一种增效剂（VX-770）联合使用，在囊性纤维化（CF）治疗方面取得了显著疗效，但由最常见的突变F508del引起的稳定性和转运缺陷并未完全得到纠正。因此，仍需要更有效的CFTR校正剂。我们采用计算机模拟和分子建模方法来设计和探究新型系列CFTR校正剂（a-c）的结合位点。基于结构的研究使我们能够设计并合成新型的I类（b系列）和II类（a系列）调节剂。因此，I类调节剂的活性依赖于与Met152、Phe81、Phe191、Trp361的相互作用。II类校正剂的设计可通过涉及Gln1291或Val1288的NBD2-配体氢键来实现。此外，还提出了具有假定双重校正能力（2c）和II类校正行为（1c）的c类化合物。在F508del-CFTR CFBE细胞和原代鼻上皮细胞中的功能测定表明，14种化合物中有8种可作为CFTR校正剂，且F508del-CFTR的挽救水平与CFBE细胞中VX-809或VX-445处理后测得的水平相当。通过基于分子对接研究和作用机制的合理筛选，我们表明靶向CFTR不同结构域的化合物组合（7a + 1b和2a + 2b）可在CFBE细胞系和原代鼻细胞中累加/协同挽救F508del-CFTR功能。我们的研究表明，通过计算机模拟和体外方法来开发和研究新型CFTR校正剂的作用机制，可能是优化联合校正剂疗法以协同挽救突变CFTR的一种工具。
