Positioning High-Throughput CETSA in Early Drug Discovery through Screening against B-Raf and PARP1.

Autor:	Shaw J; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Dale I; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Hemsley P; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Leach L; 2 Hit Discovery, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Alderley Park, UK., Dekki N; 3 Pelago Bioscience AB, Solna, Sweden., Orme JP; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Talbot V; 4 Mechanistic Biology & Profiling, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Narvaez AJ; 4 Mechanistic Biology & Profiling, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Bista M; 5 Structure, Biophysics & Fragment Based Lead Generation, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK., Martinez Molina D; 3 Pelago Bioscience AB, Solna, Sweden., Dabrowski M; 3 Pelago Bioscience AB, Solna, Sweden., Main MJ; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK.; 6 Medicines Discovery Catapult, Mereside, Alderley Park, UK., Gianni D; 1 Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK.
Jazyk:	angličtina
Zdroj:	SLAS discovery : advancing life sciences R & D [SLAS Discov] 2019 Feb; Vol. 24 (2), pp. 121-132. Date of Electronic Publication: 2018 Dec 13.
DOI:	10.1177/2472555218813332
Abstrakt:	Methods to measure cellular target engagement are increasingly being used in early drug discovery. The Cellular Thermal Shift Assay (CETSA) is one such method. CETSA can investigate target engagement by measuring changes in protein thermal stability upon compound binding within the intracellular environment. It can be performed in high-throughput, microplate-based formats to enable broader application to early drug discovery campaigns, though high-throughput forms of CETSA have only been reported for a limited number of targets. CETSA offers the advantage of investigating the target of interest in its physiological environment and native state, but it is not clear yet how well this technology correlates to more established and conventional cellular and biochemical approaches widely used in drug discovery. We report two novel high-throughput CETSA (CETSA HT) assays for B-Raf and PARP1, demonstrating the application of this technology to additional targets. By performing comparative analyses with other assays, we show that CETSA HT correlates well with other screening technologies and can be applied throughout various stages of hit identification and lead optimization. Our results support the use of CETSA HT as a broadly applicable and valuable methodology to help drive drug discovery campaigns to molecules that engage the intended target in cells.
Databáze:	MEDLINE
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