Development of a novel secondary phenotypic screen to identify hits within the mycobacterial protein synthesis pipeline.
RNA polymerase
mCherry
mycobacteria
ribosome
transcription
translation
Journal
FASEB bioAdvances
ISSN: 2573-9832
Titre abrégé: FASEB Bioadv
Pays: United States
ID NLM: 101733210
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
17
04
2020
revised:
17
04
2020
accepted:
31
07
2020
entrez:
22
10
2020
pubmed:
23
10
2020
medline:
23
10
2020
Statut:
epublish
Résumé
Whole-cell phenotypic screening is the driving force behind modern anti-tubercular drug discovery efforts. Focus has shifted from screening for bactericidal scaffolds to screens incorporating target deconvolution. Target-based screening aims to direct drug discovery toward known effective targets and avoid investing resources into unproductive lines of enquiry. The protein synthesis pipeline, including RNA polymerase and the ribosome, is a clinically proven target in Using The assay was validated using known inhibitors of protein synthesis to show a dose-dependent reduction in mCherry fluorescence. This was expanded to a proprietary screen of hypothetical protein synthesis hits and modified to include quantitative viability measurement of cells using resazurin. Following the success of the proprietary screen, a larger scale screen of the GlaxoSmithKline anti-tubercular library containing 2799 compounds was conducted. Combined single shot and dose-response screening yielded 18 hits, 0.64% of all screened compounds.
Sections du résumé
BACKGROUND
BACKGROUND
Whole-cell phenotypic screening is the driving force behind modern anti-tubercular drug discovery efforts. Focus has shifted from screening for bactericidal scaffolds to screens incorporating target deconvolution. Target-based screening aims to direct drug discovery toward known effective targets and avoid investing resources into unproductive lines of enquiry. The protein synthesis pipeline, including RNA polymerase and the ribosome, is a clinically proven target in
METHODS
METHODS
Using
RESULTS
RESULTS
The assay was validated using known inhibitors of protein synthesis to show a dose-dependent reduction in mCherry fluorescence. This was expanded to a proprietary screen of hypothetical protein synthesis hits and modified to include quantitative viability measurement of cells using resazurin.
CONCLUSION
CONCLUSIONS
Following the success of the proprietary screen, a larger scale screen of the GlaxoSmithKline anti-tubercular library containing 2799 compounds was conducted. Combined single shot and dose-response screening yielded 18 hits, 0.64% of all screened compounds.
Identifiants
pubmed: 33089076
doi: 10.1096/fba.2020-00022
pii: FBA21161
pmc: PMC7566049
doi:
Types de publication
Journal Article
Langues
eng
Pagination
600-612Subventions
Organisme : Medical Research Council
ID : MR/R001154/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/S000542/1
Pays : United Kingdom
Informations de copyright
©2020 The Authors. FASEB BioAdvances published by The Federation of American Societies for Experimental Biology.
Déclaration de conflit d'intérêts
The authors declare no competing financial interests.
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