Identification of G-quadruplex DNA/RNA binders: Structure-based virtual screening and biophysical characterization.

BACKGROUND

Recent findings demonstrated that, in mammalian cells, telomere DNA (Tel) is transcribed into telomeric repeat-containing RNA (TERRA), which is involved in fundamental biological processes, thus representing a promising anticancer target. For this reason, the discovery of dual (as well as selective) Tel/TERRA G-quadruplex (G4) binders could represent an innovative strategy to enhance telomerase inhibition.

METHODS

Initially, docking simulations of known Tel and TERRA active ligands were performed on the 3D coordinates of bimolecular G4 Tel DNA (Tel) and TERRA (TERRA). Structure-based pharmacophore models were generated on the best complexes and employed for the virtual screening of ~257,000 natural compounds. The 20 best candidates were submitted to biophysical assays, which included circular dichroism and mass spectrometry at different K concentrations.

RESULTS

Three hits were here identified and characterized by biophysical assays. Compound 7 acts as dual Tel/TERRA G4-ligand at physiological KCl concentration, while hits 15 and 17 show preferential thermal stabilization for Tel DNA. The different molecular recognition against the two targets was also discussed.

CONCLUSIONS

Our successful results pave the way to further lead optimization to achieve both increased selectivity and stabilizing effect against TERRA and Tel DNA G4s.

GENERAL SIGNIFICANCE

The current study combines for the first time molecular modelling and biophysical assays applied to bimolecular DNA and RNA G4s, leading to the identification of innovative ligand chemical scaffolds with a promising anticancer profile. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.