Advancing Target Validation and Ligandability Assessment: Fragment Screening via ¹⁹F NMR Spectroscopy.

Area of Science:

• Medicinal Chemistry
• Biophysics
• Structural Biology

Background:

• Precise target validation and ligandability assessment are critical in drug discovery.
• Fragment-based screening (FBS) is a key strategy for identifying novel drug leads.
• Fluorine-19 (19F) NMR spectroscopy offers unique advantages for biomolecular studies.

Purpose of the Study:

• To introduce an advanced protocol using 19F NMR for fragment-based screening.
• To demonstrate the utility of this method in distinguishing fragment hits and elucidating binding interactions.
• To highlight its application in early-stage drug discovery, focusing on target validation and ligandability assessment.

Main Methods:

• Utilized fluorine-19 (19F) NMR spectroscopy for fragment-based screening.
• Applied the protocol to assess fragment binding interactions at millimolar concentrations.
• Employed 19F NMR's high signal dispersion, background-free spectra, and sensitivity to protein binding.

Main Results:

• Successfully distinguished fragment hits from screening experiments.
• Elucidated the binding interactions of identified fragments.
• Demonstrated the method's effectiveness even at millimolar fragment concentrations.

Conclusions:

• The developed 19F NMR protocol is a potent tool for fragment-based screening in early-stage drug discovery.
• This technique facilitates robust target validation and ligandability assessment.
• It provides valuable insights into potential drug targets and their binding challenges from the project's inception.