Fragment screening by weak affinity chromatography: comparison with established techniques for screening against...

Area of Science:

• Drug discovery and development
• Biophysical techniques
• Fragment-based lead discovery (FBLD)

Background:

• Fragment-based lead discovery (FBLD) is crucial for drug development.
• Established methods like NMR, SPR, and X-ray crystallography are used for fragment screening.
• New technologies are needed to improve the sensitivity and reliability of fragment detection.

Purpose of the Study:

• To compare the efficacy of Weak Affinity Chromatography (WAC) with established fragment screening methods.
• To evaluate WAC's performance in identifying fragments that bind to the HSP90 ATPase domain.
• To assess the stability and reproducibility of WAC columns.

Main Methods:

• Screening of 111 fragments against the HSP90 ATPase domain using WAC, Nuclear Magnetic Resonance (NMR), Surface Plasmon Resonance (SPR), X-ray crystallography, thermal shift, and fluorescence polarization (FP).
• Isothermal Titration Calorimetry (ITC) was used for hit validation.
• WAC column stability and reproducibility were assessed over time.

Main Results:

• WAC demonstrated high hit correlation with established methods, showing 88% with NMR and 83% with SPR.
• WAC columns exhibited 90% reproducibility even after 207 days of storage.
• The study found good correlation between WAC and other fragment screening technologies.

Conclusions:

• Weak Affinity Chromatography (WAC) is a validated and effective technology for fragment screening in drug discovery.
• WAC offers comparable performance to established methods like NMR and SPR for hit identification.
• WAC is a sensitive, reliable, and stable method for identifying potential drug leads.