Efficient affinity ranking of fluorinated ligands by 19F NMR: CSAR and FastCSAR

J Biomol NMR. 2020 Nov;74(10-11):579-594. doi: 10.1007/s10858-020-00325-x. Epub 2020 Jun 16.

Abstract

Fluorine NMR has recently gained high popularity in drug discovery as it allows efficient and sensitive screening of large numbers of ligands. However, the positive hits found in screening must subsequently be ranked according to their affinity in order to prioritize them for follow-up chemistry. Unfortunately, the primary read-out from the screening experiments, namely the increased relaxation rate upon binding, is not proportional to the affinity of the ligand, as it is polluted by effects such as exchange broadening. Here we present the method CSAR (Chemical Shift-anisotropy-based Affinity Ranking) for reliable ranking of fluorinated ligands by NMR, without the need of isotope labeled protein, titrations or setting up a reporter format. Our strategy is to produce relaxation data that is directly proportional to the binding affinity. This is achieved by removing all other contributions to relaxation as follows: (i) exchange effects are efficiently suppressed by using high power spin lock pulses, (ii) dipolar relaxation effects are approximately subtracted by measuring at two different magnetic fields and (iii) differences in chemical shift anisotropy are normalized using calculated values. A similar ranking can be obtained with the simplified approach FastCSAR that relies on a measurement of a single relaxation experiment at high field (preferably > 600 MHz). An affinity ranking obtained in this simple way will enable prioritizing ligands and thus improve the efficiency of fragment-based drug design.

Keywords: Affinity ranking; Chemical shift anisotropy (CSA); Density functional theory (DFT) calculations; Drug discovery; Fluorine NMR; Fragment-based drug design (FBDD); Spin lock.

MeSH terms

  • Anisotropy
  • Density Functional Theory
  • Drug Design
  • Drug Discovery / methods*
  • Fluorine / chemistry*
  • Ligands
  • Magnetic Fields
  • Magnetic Resonance Spectroscopy / methods*
  • Proteins / chemistry*

Substances

  • Ligands
  • Proteins
  • Fluorine