Fragment screening by biochemical assay.

Area of Science:

• Biochemistry
• Medicinal Chemistry
• Molecular Biology

Background:

• High concentration biochemical assays are an effective strategy for identifying weak binding fragment molecules.
• These assays serve as novel starting points for medicinal chemistry programs.
• They offer a viable alternative to common fragment screening methods like NMR and high-throughput X-ray crystallography.

Purpose of the Study:

• To evaluate the efficacy of high concentration biochemical assays in fragment-based drug discovery.
• To present a sensitive and reproducible screening method using a portfolio of single-molecule fluorescence correlation spectroscopy techniques.
• To demonstrate the utility of X-ray crystallography in determining the binding modes of identified active fragments.

Main Methods:

• Utilized high concentration biochemical assays for fragment screening.
• Employed a portfolio of single-molecule fluorescence correlation spectroscopy (smFCS) detection techniques.
• Applied X-ray crystallography to elucidate the binding modes of active fragments.

Main Results:

• Demonstrated the high reproducibility and sensitivity of fragment screening using smFCS.
• Successfully identified weak binding fragment molecules using high concentration biochemical assays.
• Confirmed the binding modes of identified fragments through X-ray crystallography.

Conclusions:

• High concentration biochemical assays combined with sensitive detection techniques like smFCS are effective for fragment-based drug discovery.
• This approach provides a robust alternative to conventional screening methods.
• The identified fragments represent valuable starting points for developing new therapeutics.