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Updated: May 21, 2025
Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology
Published on: May 2, 2025
Affinity-based protein profiling of MDM2 inhibitor Navtemadlin
Amrita Date1, Archie Wall1, Peiyu Zhang2,3
1Department of Chemistry, Molecular Sciences Research Hub, Imperial College London 82 Wood Lane London W12 0BZ UK a.barnard@imperial.ac.uk.
Abstract:
Navtemadlin is a potent inhibitor of the p53-MDM2 protein-protein interaction, which plays a critical role in the proliferation of p53-wildtype tumours. Whilst Navtemadlin has progressed to multiple Phase III clinical trials in oncology, little has been disclosed regarding its selectivity for MDM2 in cells. Here, we report the synthesis and validation of photoactivatable clickable probes of Navtemadlin, and their application to de novo target discovery for Navtemadlin through affinity-based protein profiling. MDM2 was robustly identified as the main target, across two cell lines, using two distinct probe designs. While off-targets were identified, these were not consistent across cell lines and probe designs, consistent with a high degree of selectivity for the target protein. Whole proteome profiling experiments across different time points confirmed p53-mediated phenotypic activity and revealed novel expression patterns for key proteins in the p53 pathway.
Insights
Navtemadlin effectively inhibits the p53-MDM2 interaction, crucial for p53-wildtype tumor growth. Affinity-based profiling confirmed MDM2 as the primary target, demonstrating Navtemadlin
Area of Science:
- Oncology
- Molecular Biology
- Chemical Biology
Background:
- Navtemadlin is a p53-MDM2 protein-protein interaction inhibitor.
- Its selectivity profile in cellular environments remains largely undisclosed.
- The p53-MDM2 interaction is critical for p53-wildtype tumor proliferation.
Purpose of the Study:
- To synthesize and validate photoactivatable clickable probes for Navtemadlin.
- To perform de novo target discovery using affinity-based protein profiling.
- To assess the cellular selectivity of Navtemadlin.
Main Methods:
- Synthesis of photoactivatable clickable Navtemadlin probes.
- Affinity-based protein profiling (chemical proteomics).
- Whole proteome profiling across different time points.
Main Results:
- MDM2 was robustly identified as the primary target across two cell lines and two probe designs.
- Off-target interactions were inconsistent, suggesting high target selectivity.
- p53-mediated phenotypic activity was confirmed, with novel p53 pathway protein expression patterns identified.
Conclusions:
- Navtemadlin exhibits high selectivity for MDM2 in cellular systems.
- Photoactivatable probes are effective tools for target deconvolution and profiling.
- The study provides insights into Navtemadlin's mechanism of action and potential biomarkers.
