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A strategy for high-throughput assay development using leads derived from nuclear magnetic resonance-based screening.
Philip J Hajduk1, Stephen F Betz, Jamey Mack
1Global Pharmaceutical Research and Development, Abbott Laboratories, Abbott Park, IL, USA.
Journal of Biomolecular Screening
|November 6, 2003
Summary
This study introduces a novel nuclear magnetic resonance (NMR) spectroscopy method to discover drug leads for unknown protein targets. This approach enables rapid development of high-throughput screening assays for drug discovery.
Area of Science:
- Biochemistry
- Structural Biology
- Drug Discovery
Background:
- Developing high-throughput screening (HTS) assays for proteins with unknown functions presents a significant challenge in drug discovery.
- Nuclear Magnetic Resonance (NMR) spectroscopy offers a biophysical method to identify ligand-protein interactions without prior functional knowledge of the target.
Purpose of the Study:
- To present a strategy for developing HTS assays against protein targets of unknown function using NMR spectroscopy.
- To demonstrate the application of this strategy in creating an HTS assay for HI-0033, a conserved protein from Haemophilus influenzae.
Main Methods:
- Utilized NMR spectroscopy to screen a library of compounds for binding to the target protein HI-0033.
- Synthesized fluorescent analogs of identified binders for use in HTS.
- Developed a fluorescence polarization (FP) assay for HTS and structure-activity relationship (SAR) studies.
Main Results:
- Identified adenosine as a binder to HI-0033 using NMR.
- Developed fluorescent analogs of adenosine that bind HI-0033 with submicromolar affinity.
- Established a robust FP assay suitable for HTS and lead optimization of HI-0033 inhibitors.
Conclusions:
- The described NMR-based strategy is effective for discovering ligands and developing HTS assays for proteins of unknown function.
- This approach facilitates the identification of novel chemical matter and accelerates the drug discovery process for uncharacterized targets.
- The developed assay for HI-0033 enables further investigation into its biological role and potential as a therapeutic target.