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Nat Commun. 2016 Mar 24;7:11040. doi: 10.1038/ncomms11040.

CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil.

Author information

1
Laboratories for Chemical Biology, Karolinska Institutet, Science for Life Laboratory Stockholm, Division of Translational Medicine &Chemical Biology, Department of Medical Biochemistry &Biophysics, Karolinska Institutet, Tomtebodavägen 23A, Solna 171 65, Sweden.
2
Department of Medical Biochemistry &Biophysics, Division of Biophysics, Karolinska Institutet, Scheeles väg 2, Stockholm 171 77, Sweden.
3
School of Biological Sciences, Nanyang Technological University, 61 Biopolis Drive (Proteos), Singapore 138673, Singapore.
4
Department of Pharmacy, Uppsala University, BMC, Box 580, Uppsala SE-751 23, Sweden.
5
School of Biological Sciences, Nanyang Technological University, SBS-04s-45, 60 Nanyang Drive, Singapore 639798, Singapore.
6
Uppsala University Drug Optimization and Pharmaceutical Profiling Platform (UDOPP), Department of Pharmacy, Uppsala University, BMC, Box 580, Uppsala SE-751 23, Sweden.
7
Science for Life Laboratory Drug Discovery and Development platform, Uppsala University, Uppsala SE-751 23, Sweden.
8
Institute of Cellular and Molecular Biology, ASTAR, 61 Biopolis Drive (Proteos), Singapore 138673, Singapore.

Abstract

Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular target binding, exemplified by human thymidylate synthase (TS). The screen selected accurately for all the tested known drugs acting on TS. We also identified TS inhibitors with novel chemistry and marketed drugs that were not previously known to target TS, including the DNA methyltransferase inhibitor decitabine. By following the cellular uptake and enzymatic conversion of known drugs we correlated the appearance of active metabolites over time with intracellular target engagement. These data distinguished a much slower activation of 5-fluorouracil when compared with nucleoside-based drugs. The approach establishes efficient means to associate drug uptake and activation with target binding during drug discovery.

PMID:
27010513
PMCID:
PMC4820820
DOI:
10.1038/ncomms11040
[Indexed for MEDLINE]
Free PMC Article

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