¹⁹F chemical library and ¹⁹F-NMR for a weakly bound complex structure

Area of Science:

• Biochemistry
• Structural Biology
• Medicinal Chemistry

Background:

• Fragment-based drug discovery (FBDD) is a powerful technique utilizing small molecules (<300 Da).
• Low affinity of hit compounds in FBDD presents challenges in analyzing protein-compound interactions.
• Developing efficient screening and analysis methods is crucial for FBDD success.

Purpose of the Study:

• To develop and validate a novel 19F-NMR screening method for FBDD.
• To optimize a 19F chemical library for high-throughput screening of soluble monomeric molecules.
• To enable precise structural modeling of protein-weak binder complexes.

Main Methods:

• Optimization of a 19F chemical library with highly soluble monomeric molecules.
• Application of a 19F-NMR screening method to four diverse proteins, including kinases and a membrane protein.
• Validation of hit compounds using protein thermal shift analysis, 1H-15N HSQC NMR, and isothermal titration calorimetry.
• Utilizing 1H and 19F saturation transfer difference experiments for precise structural determination.

Main Results:

• Successful application of the 19F-NMR screening method across multiple protein targets.
• Accurate determination of dissociation constants for weak binders.
• Generation of highly precise structural models for protein-compound complexes.
• Demonstration of the method's efficacy in overcoming challenges associated with low-affinity binders.

Conclusions:

• The developed 19F-NMR screening method, combined with an optimized 19F chemical library, effectively facilitates the analysis of weak protein-compound interactions in FBDD.
• This approach enables the structural modeling of complexes formed between low-affinity binders and their target proteins.
• The study provides a robust platform for advancing drug discovery efforts through enhanced fragment screening and structural analysis.