title: Discovery of New E-Selectin Inhibitors by Virtual Screening, Fluorescence Binding Assays, and STD NMR Experiments.
abstract: E-selectin is an endothelial protein that participates in the adhesion of metastatic cancer cells, and is therefore a relevant target for antitumor therapeutic intervention. In this work, virtual screening was used to identify new E-selectin inhibitors from a subset of drug-like molecules retrieved from the ZINC database, including the physiological ligand sLe(x) as reference structure (PDB ID: 1G1T). Four hits were chosen and subjected to molecular dynamics simulations and fluorescence binding assays, which led to the determination of experimental dissociation constants between 333 and 1012 μm. The candidate with the highest affinity was studied by saturation transfer difference (STD) NMR experiments and complete relaxation and conformational exchange matrix analysis of saturation transfer (CORCEMA-ST), aimed at identifying the preferable binding mode with E-selectin. Our results revealed that this new inhibitor binds more strongly than sLe(x) in the E-selectin binding site, in good agreement with simulation predictions. These properties will prove valuable for the future design of drugs that target E-selectin.
authors.affiliation: Department of Organic Chemistry, Universidad de Concepción, Casilla 160-C, Concepción, Chile.
authors.forename: Pabla A
authors.initials: PA
authors.lastname: Barra
authors.affiliation: Department of Chemical Sciences, Faculty of Exact Sciences, Universidad Andres Bello, Autopista Concepción-Talcahuano 7100, Talcahuano, Chile.
authors.forename: Verónica A
authors.initials: VA
authors.lastname: Jiménez
authors.affiliation: Instituto Universitario de Bioorgánica "A. González", Universidad de La Laguna, Avenida Astrofísico Francisco Sánchez, 38206, La Laguna, Tenerife, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Gavin
authors.affiliation: Instituto Universitario de Bioorgánica "A. González", Universidad de La Laguna, Avenida Astrofísico Francisco Sánchez, 38206, La Laguna, Tenerife, Spain.
authors.forename: Antonio H
authors.initials: AH
authors.lastname: Daranas
authors.affiliation: Department of Organic Chemistry, Universidad de Concepción, Casilla 160-C, Concepción, Chile. jalderet@udec.cl.
authors.forename: Joel B
authors.initials: JB
authors.lastname: Alderete
mesh_terms: D001555:Benzene Derivatives / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D019040:E-Selectin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D056004:Molecular Dynamics Simulation; D010793:Phthalazines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence
chemical_list: D001555:Benzene Derivatives; D019040:E-Selectin; D008024:Ligands; D010793:Phthalazines; C015763:cumene
keywords: E-selectin; NMR spectroscopy; drug design; molecular dynamics; virtual screening