High throughput screening of 0.5 million compounds against CRAF using Alpha CETSAⓇ

Tschapalda, Kirsten; Blackett, Carolyn; Ivanov, Delyan P.; Plant, Darren; Shaw, J.C.; Thomas, Andrew P.; Packer, Martin J.; Arnold, Laurence H.; Holdgate, Geoffrey A.

doi:10.1016/j.slasd.2023.01.006

Cited by 9 publications

(5 citation statements)

References 18 publications

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“…Идея была выведена на совершенно новый уровень с разработкой метода микромасштабного термофореза (NanoTemper Technologies GmbH, Германия), позволяющего отслеживать изменения «водного щита», окружающего белок, при связывании соединения-лиганда. Недавнее применение модернизированного метода для скрининга библиотеки объемом в полмиллиона соединений было описано в [23]. Другой подход HTS -масс-спектрометрия с аффинной селекцией (AS-MS), был использован компанией «Merck» для идентификации ингибиторов пролилгидроксилазы HIF, работающих не в активном центре фермента [24].…”

Section: скрининг по принципу конкурентного замещения белка -преимуще...unclassified

High Throughput Screening in Drug Discovery: Problems And Solutions

Hushpulian,

Gaisina,

Nikulin

et al. 2024

Lomonosov Chemistry Journal

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World-wide introduction of high throughput screening (HTS) methods in drug discovery research did not result in the increased number of novel medications on the market. We discuss novel trends in drug discovery that came from the understanding that majority of diseases are multifactorial and that one enzyme has many protein substrates. Hence, new approaches are focused on development of drugs, which (1) trigger survival pathways to return the organism to homeostatic balance, and (2) inhibit enzymes modifying histones or transcription factors not at the active site, but by displacement of protein substrates from the enzyme complexes. A good example for both approaches comes from the development of activators of antioxidant defense. We analyze and illustrate problems of commonly used in vitro HTS assays, and briefl y discuss advantages and limitations of small animal models. The novel approaches are complementary to the standard HTS and do not substitute for testing in mammals. Development of transgenic reporter mice to monitor drug effects by means of in vivo imaging is extremely promising to select proper dosage and administration regimes for full-range PK studies.

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Section: скрининг по принципу конкурентного замещения белка -преимуще...unclassified

High Throughput Screening in Drug Discovery: Problems And Solutions

Hushpulian,

Gaisina,

Nikulin

et al. 2024

Lomonosov Chemistry Journal

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“…18,19 In a recent proof-of-concept study by AstraZeneca, CETSA was used for primary screening of a 500K compound library against the well-validated oncology target RAF protooncogene serine/threonine-protein kinase CRAF. 20 The developed CETSA HT assay proved to be robust, with a very low frequency of false positives and low susceptibility to PAINS. False positive hits in a CETSA screen could be caused by aggregation or assay interference by the compound, and it is important to carefully triage the hits and include a counter screen in the screening funnel.…”

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confidence: 94%

“…The CETSA HT format has mainly been used for screening of focused compound libraries and in hit confirmation efforts. , In a recent proof-of-concept study by AstraZeneca, CETSA was used for primary screening of a 500K compound library against the well-validated oncology target RAF proto-oncogene serine/threonine-protein kinase CRAF . The developed CETSA HT assay proved to be robust, with a very low frequency of false positives and low susceptibility to PAINS.…”

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confidence: 99%

“…This was confirmed by Western blot-based CETSA. The thermal stabilization was found to occur at an order of magnitude higher than the autophagy inhibitory activity, consistent with the requirement of high compound concentration in CETSA to detect thermal stability changes …”

mentioning

confidence: 99%

“…The thermal stabilization was found to occur at an order of magnitude higher than the autophagy inhibitory activity, consistent with the requirement of high compound concentration in CETSA to detect thermal stability changes. 20 Wilke et al discovered a Sigma 1 receptor (S1) antagonist by using a morphological screen based on cell painting followed by proteome-wide CETSA MS for identifying the primary target of the cellular response. 29 In the study, 10,000 compounds were profiled in a cell painting assay to identify bioactive compounds.…”

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confidence: 99%

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A Shift in Thinking: Cellular Thermal Shift Assay-Enabled Drug Discovery

Caballero

Lundgren

2023

ACS Med. Chem. Lett.

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A decade has passed since the cellular thermal shift assay (CETSA) was introduced to the drug discovery community. Over the years, the method has guided numerous projects by providing insights about, for example, target engagement, lead generation, target identification, lead optimization, and preclinical profiling. With this Microperspective, we intend to highlight recently published applications of CETSA and how the data generated can enable efficient decision-making and prioritization throughout the drug discovery and development value chain.

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