title: Enhancement of premature stop codon readthrough in the CFTR gene by Ataluren (PTC124) derivatives.
abstract: Premature stop codons are the result of nonsense mutations occurring within the coding sequence of a gene. These mutations lead to the synthesis of a truncated protein and are responsible for several genetic diseases. A potential pharmacological approach to treat these diseases is to promote the translational readthrough of premature stop codons by small molecules aiming to restore the full-length protein. The compound PTC124 (Ataluren) was reported to promote the readthrough of the premature UGA stop codon, although its activity was questioned. The potential interaction of PTC124 with mutated mRNA was recently suggested by molecular dynamics (MD) studies highlighting the importance of H-bonding and stacking π-π interactions. To improve the readthrough activity we changed the fluorine number and position in the PTC124 fluoroaryl moiety. The readthrough ability of these PTC124 derivatives was tested in human cells harboring reporter plasmids with premature stop codons in H2BGFP and FLuc genes as well as in cystic fibrosis (CF) IB3.1 cells with a nonsense mutation. Maintaining low toxicity, three of these molecules showed higher efficacy than PTC124 in the readthrough of the UGA premature stop codon and in recovering the expression of the CFTR protein in IB3.1 cells from cystic fibrosis patient. Molecular dynamics simulations performed with mutated CFTR mRNA fragments and active or inactive derivatives are in agreement with the suggested interaction of PTC124 with mRNA.
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy; Istituto EuroMediterraneo di Scienza e Tecnologia (IEMEST), Via Emerico Amari 123, 90139 Palermo, Italy.
authors.forename: Ivana
authors.initials: I
authors.lastname: Pibiri
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Lentini
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Melfi
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Gallucci
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy; Istituto EuroMediterraneo di Scienza e Tecnologia (IEMEST), Via Emerico Amari 123, 90139 Palermo, Italy. Electronic address: andrea.pace@unipa.it.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pace
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Spinello
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy; Istituto EuroMediterraneo di Scienza e Tecnologia (IEMEST), Via Emerico Amari 123, 90139 Palermo, Italy.
authors.forename: Giampaolo
authors.initials: G
authors.lastname: Barone
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 17, 90128 Palermo, Italy; Centro di OncoBiologia Sperimentale (COBS), Via San Lorenzo Colli, 90145 Palermo, Italy. Electronic address: aldo.dileonardo@unipa.it.
authors.forename: Aldo
authors.initials: A
authors.lastname: Di Leonardo
mesh_terms: D018389:Codon, Nonsense / Q000235:genetics*; D003550:Cystic Fibrosis / Q000188:drug therapy* / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000096:biosynthesis* / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D056004:Molecular Dynamics Simulation; D015394:Molecular Structure; D009154:Mutation; D010069:Oxadiazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: C505032:CFTR protein, human; D018389:Codon, Nonsense; D010069:Oxadiazoles; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C515878:ataluren
keywords: CFTR gene; Cystic fibrosis; Fluorinated oxadiazole; Green fluorescent protein; Nonsense mutation; PTCs readthrough