Exploitation of a novel biosensor based on the full-length human F508del-CFTR with computational studies, biochemical and biological assays for the characterization of a new Lumacaftor/Tezacaftor analogue

Abstract Cystic fibrosis (CF) is mainly caused by the mutation F508del of the cystic fibrosis transmembrane conductance regulator (CFTR) that is thus retained in the endoplasmic reticulum and degraded. New drugs able to rescue F508del-CFTR trafficking and activity are eagerly awaited, a goal that requires the availability of computational and experimental models closely resembling the F508del-CFTR structure and environment in vivo. Here we describe the development of a biosensor based on F508del-CFTR in a lipid environment that proved to be endowed with a wider analytical potential in respect to the previous CFTR-based biosensors. Integrated with an appropriate computational model of the whole human F508del-CFTR in lipid environment and CFTR stability and functional assays, the new biosensor allowed the identification and characterization at the molecular level of the binding modes of some known F508del-CFTR-rescuing drugs and of a new aminoarylthiazole-Lumacaftor/Tezacaftor hybrid derivative endowed with promising F508del-CFTR-binding and rescuing activity.