2. Academic Validation
  3. Ligand-Based Competition Binding by Real-Time 19F NMR in Human Cells

Ligand-Based Competition Binding by Real-Time 19F NMR in Human Cells

  • J Med Chem. 2024 Jan 25;67(2):1115-1126. doi: 10.1021/acs.jmedchem.3c01600.
Enrico Luchinat 1 2,  Letizia Barbieri 2,  Ben Davis 3,  Paul A Brough 3,  Matteo Pennestri 4,  Lucia Banci 2 5 6
Affiliations

Affiliations

  • 1 Dipartimento di Scienze e Tecnologie Agro-Alimentari, Alma Mater Studiorum─Università di Bologna, Piazza Goidanich 60, Cesena 47521, Italy.
  • 2 Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine─CIRMMP, Via Luigi Sacconi 6, Sesto Fiorentino 50019, Italy.
  • 3 Vernalis Research, Granta Park, Great Abington, Cambridge CB21 6GB, U.K.
  • 4 Pharmaceutical Business Unit, Bruker UK Limited, Banner Lane, Coventry CV4 9GH, U.K.
  • 5 Centro di Risonanze Magnetiche─CERM, Università degli Studi di Firenze, Via Luigi Sacconi 6, Sesto Fiorentino 50019, Italy.
  • 6 Dipartimento di Chimica, Università degli Studi di Firenze, Via della Lastruccia 3, Sesto Fiorentino 50019, Italy.
PMID: 38215028 DOI: 10.1021/acs.jmedchem.3c01600
Abstract

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.

Figures