posted on 2024-01-12, 18:08authored byEnrico Luchinat, Letizia Barbieri, Ben Davis, Paul A. Brough, Matteo Pennestri, Lucia Banci
The development of more effective drugs requires knowledge
of their
bioavailability and binding efficacy directly in the native cellular
environment. In-cell nuclear magnetic resonance (NMR) spectroscopy
is a powerful tool for investigating ligand–target interactions
directly in living cells. However, the target molecule may be NMR-invisible
due to interactions with cellular components, while observing the
ligand by 1H NMR is impractical due to the cellular background.
Such limitations can be overcome by observing fluorinated ligands
by 19F in-cell NMR as they bind to the intracellular target.
Here we report a novel approach based on real-time in-cell 19F NMR that allows measuring ligand binding affinities in human cells
by competition binding, using a fluorinated compound as a reference.
The binding of a set of compounds toward Hsp90α was investigated.
In principle, this approach could be applied to other pharmacologically
relevant targets, thus aiding the design of more effective compounds
in the early stages of drug development.