PTC124 derivatives as a novel approach to improve the readthrough of premature stop codons in the CFTR gene

Progetto: Research project

Dettagli progetto


The presence in mRNA of premature stop codons (PTCs) results in protein truncation responsible for inherited (genetic) diseases. A well-known example is cystic fibrosis (CF), where approximately 10% (worldwide) of patients have nonsense mutations in the CF trans-membrane regulator (CFTR) gene. Pharmacological approaches aimed to rescue protein function have been proposed to directly overcome nonsense mutations. PTC124 (Ataluren) a small molecule that mimic the activity of aminoglycosides has been suggested to allow PTCs readthrough. However, despite the results obtained from "in vitro" and "in vivo" experiments as well the advanced clinical trials done with PTC124, some caveats exist. In fact PTC124 has a lower activity against ochre and amber non sense mutations. There is no general consensus about its mechanism of action and very high doses have to be administered to reach working blood concentrations. The project is aimed to design and synthesize new molecules, alternatives or complementary to PTC124, able to promote the PTCs readthrough and with better bioavailability. Heterocycles possessing the geometrical requirements to match the hydrogen bonding of the nonsense stop codon in the mRNA will be synthesized. Ability of the new molecules to promote the readthrough will be tested in cells transfected with pBOS-H2BGFP plasmid harboring PTCs and in CF epithelial cells (IB3-1). Cells will be treated with PTC124, as a control, and with different PTC124 derivatives. The effect of these compounds will be monitored by fluorescence microscopy and at molecular level by immunoblotting and Real time PCR. First year: design and synthesis of 24 new PTC124 derivatives; additional reporter vector mutagenesis, cell transfection and treatment with new synthesized molecules. Second year: synthesis of selected derivatives with improved bioavailability as suggested by biological data; treatments on transfected cells with reporter vector and their validation on IB3-1 cells.
Data di inizio/fine effettiva1/1/11 → …


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