Assay strategies for the discovery and validation of therapeutics targeting Brugia pahangi Hsp90

Taldone, T., Gillan, V., Sun, W., Rodina, A., Patel, P., Maitland, K., O'Neill, K., Chiosis, G. and Devaney, E. (2010) Assay strategies for the discovery and validation of therapeutics targeting Brugia pahangi Hsp90. PLoS Neglected Tropical Diseases, 4(6), e714. (doi: 10.1371/journal.pntd.0000714) (PMID:20559560) (PMCID:PMC2886105)

Abstract

The chemotherapy of lymphatic filariasis relies upon drugs such as diethylcarbamazine and ivermectin that largely target the microfilarial stages of the parasite, necessitating continued treatment over the long reproductive life span of the adult worm. The identification of compounds that target adult worms has been a long-term goal of WHO. Here we describe a fluorescence polarization assay for the identification of compounds that target Hsp90 in adult filarial worms. The assay was originally developed to identify inhibitors of Hsp90 in tumor cells, and relies upon the ability of small molecules to inhibit the binding of fluorescently labelled geldanamycin to Hsp90. We demonstrate that the assay works well with soluble extracts of Brugia, while extracts of the free-living nematode C. elegans fail to bind the probe, in agreement with data from other experiments. The assay was validated using known inhibitors of Hsp90 that compete with geldanamycin for binding to Hsp90, including members of the synthetic purine-scaffold series of compounds. The efficacy of some of these compounds against adult worms was confirmed in vitro. Moreover, the assay is sufficiently sensitive to differentiate between binding of purine-scaffold compounds to human and Brugia Hsp90. The assay is suitable for high-throughput screening and provides the first example of a format with the potential to identify novel inhibitors of Hsp90 in filarial worms and in other parasitic species where Hsp90 may be a target.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Devaney, Professor Eileen and Gillan, Dr Victoria and O'Neill, Ms Kerry and Maitland, Ms Kirsty
• Authors: Taldone, T., Gillan, V., Sun, W., Rodina, A., Patel, P., Maitland, K., O'Neill, K., Chiosis, G., and Devaney, E.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: PLoS Neglected Tropical Diseases
• Publisher: Public Library of Science
• ISSN: 1935-2727
• ISSN (Online): 1935-2735
• Published Online: 15 June 2010
• Copyright Holders: Copyright © 2010 The Authors
• First Published: First published in PLoS Neglected Tropical Diseases 4(6):e714
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
44159	1	A comparative analysis of Hsp90 its possible role in drug resistance in nematode parasites	Eileen Devaney	Biotechnology and Biological Sciences Research Council (BBSRC)	BB/E013473/1	Infection Immunity and Inflammation Life Sciences