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High-throughput screening for Hsp90 ATPase inhibitors.
Christopher Avila1, M Kyle Hadden, Zeqiang Ma
1Department of Medicinal Chemistry, The University of Kansas, 1251 Wescoe Hall Drive, Malott 4070, Lawrence, KS 66045-7563, USA.
Bioorganic & Medicinal Chemistry Letters
|March 15, 2006
Summary
A new assay for yeast heat shock protein 90 (Hsp90) ATPase activity was validated for large-scale screening. This method identified novel Hsp90 inhibitors from approximately 10,000 compounds.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Heat shock protein 90 (Hsp90) is a crucial molecular chaperone involved in protein folding and stability.
- Dysregulation of Hsp90 activity is implicated in various diseases, including cancer, making it a therapeutic target.
- Development of robust assays for Hsp90 is essential for identifying novel inhibitors.
Purpose of the Study:
- To validate a newly developed assay for determining yeast Hsp90 ATPase activity.
- To assess the feasibility of using this assay for large-scale, high-throughput screening (HTS).
- To identify potential Hsp90 inhibitors from a diverse compound library.
Main Methods:
- A previously reported assay for yeast Hsp90 ATPase activity was employed.
- High-throughput screening of approximately 10,000 chemical compounds was conducted.
- Assay reproducibility was assessed using the Z-factor metric.
Main Results:
- The Hsp90 ATPase activity assay demonstrated high reproducibility, with an average Z-factor of 0.80.
- HTS identified 0.57% of screened compounds as Hsp90 inhibitors.
- Inhibitors exhibited half-maximal inhibitory concentrations (IC50s) below 20 microM.
Conclusions:
- The developed assay is suitable for large-scale HTS of potential Hsp90 inhibitors.
- Several novel inhibitory scaffolds targeting yeast Hsp90 were identified.
- These findings provide a foundation for further development of Hsp90-targeted therapeutics.