A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes.
Human African Trypanosomiasis is a vector-borne disease of sub-Saharan Africa that causes significant morbidity and mortality. Current therapies have many drawbacks, and there is an urgent need for new, better medicines. Ideally such new treatments should be fast-acting cidal agents that cure the di...
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ftdoajarticles:oai:doaj.org/article:51b3428b92254b6fb1b0ad98fc0b8437 2023-05-15T15:12:34+02:00 A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. Manu De Rycker Sandra O'Neill Dhananjay Joshi Lorna Campbell David W Gray Alan H Fairlamb 2012-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001932 https://doaj.org/article/51b3428b92254b6fb1b0ad98fc0b8437 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3510075?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001932 https://doaj.org/article/51b3428b92254b6fb1b0ad98fc0b8437 PLoS Neglected Tropical Diseases, Vol 6, Iss 11, p e1932 (2012) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2012 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001932 2022-12-31T13:04:59Z Human African Trypanosomiasis is a vector-borne disease of sub-Saharan Africa that causes significant morbidity and mortality. Current therapies have many drawbacks, and there is an urgent need for new, better medicines. Ideally such new treatments should be fast-acting cidal agents that cure the disease in as few doses as possible. Screening assays used for hit-discovery campaigns often do not distinguish cytocidal from cytostatic compounds and further detailed follow-up experiments are required. Such studies usually do not have the throughput required to test the large numbers of hits produced in a primary high-throughput screen. Here, we present a 384-well assay that is compatible with high-throughput screening and provides an initial indication of the cidal nature of a compound. The assay produces growth curves at ten compound concentrations by assessing trypanosome counts at 4, 24 and 48 hours after compound addition. A reduction in trypanosome counts over time is used as a marker for cidal activity. The lowest concentration at which cell killing is seen is a quantitative measure for the cidal activity of the compound. We show that the assay can identify compounds that have trypanostatic activity rather than cidal activity, and importantly, that results from primary high-throughput assays can overestimate the potency of compounds significantly. This is due to biphasic growth inhibition, which remains hidden at low starting cell densities and is revealed in our static-cidal assay. The assay presented here provides an important tool to follow-up hits from high-throughput screening campaigns and avoid progression of compounds that have poor prospects due to lack of cidal activity or overestimated potency. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 6 11 e1932 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Manu De Rycker Sandra O'Neill Dhananjay Joshi Lorna Campbell David W Gray Alan H Fairlamb A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Human African Trypanosomiasis is a vector-borne disease of sub-Saharan Africa that causes significant morbidity and mortality. Current therapies have many drawbacks, and there is an urgent need for new, better medicines. Ideally such new treatments should be fast-acting cidal agents that cure the disease in as few doses as possible. Screening assays used for hit-discovery campaigns often do not distinguish cytocidal from cytostatic compounds and further detailed follow-up experiments are required. Such studies usually do not have the throughput required to test the large numbers of hits produced in a primary high-throughput screen. Here, we present a 384-well assay that is compatible with high-throughput screening and provides an initial indication of the cidal nature of a compound. The assay produces growth curves at ten compound concentrations by assessing trypanosome counts at 4, 24 and 48 hours after compound addition. A reduction in trypanosome counts over time is used as a marker for cidal activity. The lowest concentration at which cell killing is seen is a quantitative measure for the cidal activity of the compound. We show that the assay can identify compounds that have trypanostatic activity rather than cidal activity, and importantly, that results from primary high-throughput assays can overestimate the potency of compounds significantly. This is due to biphasic growth inhibition, which remains hidden at low starting cell densities and is revealed in our static-cidal assay. The assay presented here provides an important tool to follow-up hits from high-throughput screening campaigns and avoid progression of compounds that have poor prospects due to lack of cidal activity or overestimated potency. |
format |
Article in Journal/Newspaper |
author |
Manu De Rycker Sandra O'Neill Dhananjay Joshi Lorna Campbell David W Gray Alan H Fairlamb |
author_facet |
Manu De Rycker Sandra O'Neill Dhananjay Joshi Lorna Campbell David W Gray Alan H Fairlamb |
author_sort |
Manu De Rycker |
title |
A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
title_short |
A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
title_full |
A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
title_fullStr |
A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
title_full_unstemmed |
A static-cidal assay for Trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
title_sort |
static-cidal assay for trypanosoma brucei to aid hit prioritisation for progression into drug discovery programmes. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doi.org/10.1371/journal.pntd.0001932 https://doaj.org/article/51b3428b92254b6fb1b0ad98fc0b8437 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 6, Iss 11, p e1932 (2012) |
op_relation |
http://europepmc.org/articles/PMC3510075?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001932 https://doaj.org/article/51b3428b92254b6fb1b0ad98fc0b8437 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001932 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
6 |
container_issue |
11 |
container_start_page |
e1932 |
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1766343231570706432 |