title: Bioorthogonal Probes for the Study of MDM2-p53 Inhibitors in Cells and Development of High-Content Screening Assays for Drug Discovery.
abstract: 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.
authors.affiliation: Global Discovery Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Pier Luca
authors.initials: PL
authors.lastname: D'Alessandro
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Nicole
authors.initials: N
authors.lastname: Buschmann
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Markus
authors.initials: M
authors.lastname: Kaufmann
authors.affiliation: Global Discovery Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Pascal
authors.initials: P
authors.lastname: Furet
authors.affiliation: Oncology, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Frederic
authors.initials: F
authors.lastname: Baysang
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Reto
authors.initials: R
authors.lastname: Brunner
authors.affiliation: Global Discovery Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Marzinzik
authors.affiliation: Global Discovery Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vorherr
authors.affiliation: Oncology, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Therese-Marie
authors.initials: TM
authors.lastname: Stachyra
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Johannes
authors.initials: J
authors.lastname: Ottl
authors.affiliation: Global Discovery Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Dimitrios E
authors.initials: DE
authors.lastname: Lizos
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes of Biomedical Research, Novartis Campus, Basel, Switzerland.
authors.forename: Amanda
authors.initials: A
authors.lastname: Cobos-Correa
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007211:Indoles / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D015394:Molecular Structure; D012516:Osteosarcoma / Q000188:drug therapy* / Q000473:pathology; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors*; D059010:Single-Cell Analysis; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors*
chemical_list: D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D007211:Indoles; D016159:Tumor Suppressor Protein p53; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
keywords: bioorthogonal chemistry; drug discovery; fluorescent probes; protein-protein interactions; target engagement