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

Biochim Biophys Acta Gen Subj. 2017 May;1861(5 Pt B):1329-1340. doi: 10.1016/j.bbagen.2016.12.023. Epub 2016 Dec 23.

Abstract

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 (Tel2) and TERRA (TERRA2). 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 Tel2/TERRA2 G4-ligand at physiological KCl concentration, while hits 15 and 17 show preferential thermal stabilization for Tel2 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.

Keywords: Circular dichroism; G-quadruplex DNA; Mass spectrometry; TERRA; Virtual screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / metabolism*
  • Antineoplastic Agents / pharmacology
  • Binding Sites
  • Circular Dichroism
  • DNA / chemistry
  • DNA / drug effects
  • DNA / genetics
  • DNA / metabolism*
  • Drug Design*
  • G-Quadruplexes* / drug effects
  • Guanosine / chemistry
  • Guanosine / metabolism*
  • High-Throughput Screening Assays
  • Ligands
  • Molecular Docking Simulation
  • Nucleic Acid Denaturation
  • Potassium / chemistry
  • RNA / chemistry
  • RNA / drug effects
  • RNA / genetics
  • RNA / metabolism*
  • RNA Stability
  • Spectrometry, Mass, Electrospray Ionization
  • Structure-Activity Relationship
  • Telomerase / chemistry
  • Telomerase / drug effects
  • Telomerase / genetics
  • Telomerase / metabolism*
  • Temperature

Substances

  • Antineoplastic Agents
  • Ligands
  • Guanosine
  • RNA
  • DNA
  • Telomerase
  • Potassium