title: Affinity screening using competitive binding with fluorine-19 hyperpolarized ligands.
abstract: Fluorine-19 NMR and hyperpolarization form a powerful combination for drug screening. Under a competitive equilibrium with a selected fluorinated reporter ligand, the dissociation constant (K(D)) of other ligands of interest is measurable using a single-scan Carr-Purcell-Meiboom-Gill (CPMG) experiment, without the need for a titration. This method is demonstrated by characterizing the binding of three ligands with different affinities for the serine protease trypsin. Monte Carlo simulations show that the highest accuracy is obtained when about one-half of the bound reporter ligand is displaced in the binding competition. Such conditions can be achieved over a wide range of affinities, allowing for rapid screening of non-fluorinated compounds when a single fluorinated ligand for the binding pocket of interest is known.
authors.affiliation: Department of Chemistry, Texas A & M University, College Station, TX 77843 (USA).
authors.forename: Yaewon
authors.initials: Y
authors.lastname: Kim
authors.forename: Christian
authors.initials: C
authors.lastname: Hilty
mesh_terms: D001550:Benzamidines / Q000737:chemistry / Q000378:metabolism; D001596:Benzylamines / Q000737:chemistry / Q000378:metabolism; D001667:Binding, Competitive; D005461:Fluorine / Q000737:chemistry*; D007700:Kinetics; D007976:Leupeptins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D009010:Monte Carlo Method; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
chemical_list: D001550:Benzamidines; D001596:Benzylamines; D007976:Leupeptins; D008024:Ligands; D005461:Fluorine; C030796:benzylamine; D014357:Trypsin; C032854:leupeptin; C032157:benzamidine
keywords: NMR spectroscopy; drug discovery; hyperpolarization; protein-ligand interaction