¹⁹F NMR Spectroscopy for Functional and Binding High-Throughput Screening

NMR screening has emerged as a powerful and reliable approach for identification of potential drug candidates [1]. The technique is now recognized for its impact on the drug discovery process and has become an important tool for lead identification, lead validation, and lead optimization in many pharmaceutical companies and universities [2-17]. A plethora of different NMR experiments has been proposed in the literature for performing these tasks. Two of these approaches, recently introduced, use fluorine NMR spectroscopy. FAXS (fluorine chemical shift anisotropy and exchange for screening) [18,19] and 3-FABS (three fluorine atoms for biochemical screening) [20] allow one to perform binding and functional highthroughput screening (HTS), respectively, and determine the dissociation binding constant (K_d) and the 50% mean inhibition concentration (IC₅₀) of the identified binders and inhibitors, respectively. This chapter provides an insight into the theory and practical aspects of these two experiments and presents applications to the screening of different biomolecular targets.