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Bioorthogonal Probes for the Study of MDM2‐p53 Inhibitors in Cells and Development of High‐Content Screening Assays for Drug Discovery
Author(s) -
D'Alessandro Pier Luca,
Buschmann Nicole,
Kaufmann Markus,
Furet Pascal,
Baysang Frederic,
Brunner Reto,
Marzinzik Andreas,
Vorherr Thomas,
Stachyra ThereseMarie,
Ottl Johannes,
Lizos Dimitrios E.,
CobosCorrea Amanda
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201608568
Subject(s) - drug discovery , bioorthogonal chemistry , high content screening , mdm2 , computational biology , high throughput screening , chemistry , intracellular , drug development , high resolution , small molecule , cell , drug , biochemistry , biology , combinatorial chemistry , pharmacology , apoptosis , remote sensing , click chemistry , geology
To study the behavior of MDM2‐p53 inhibitors in a disease‐relevant cellular model, we have developed and validated a set of bioorthogonal probes that can be fluorescently labeled in cells and used in high‐content screening assays. By using automated image analysis with single‐cell resolution, we could visualize the intracellular target binding of compounds by co‐localization and quantify target upregulation upon MDM2‐p53 inhibition in an osteosarcoma model. Additionally, we developed a high‐throughput assay to quantify target occupancy of non‐tagged MDM2‐p53 inhibitors by competition and to identify novel chemical matter. This approach could be expanded to other targets for lead discovery applications.