Computer-Aided Identification and Lead Optimization of Dual Murine Double Minute 2 and 4 Binders: Structure-Activity
Mariateresa Giustiniano1, Simona Daniele2, Sveva Pelliccia1
1Dipartimento di Farmacia, Università degli Studi di Napoli Federico II , Via D. Montesano 49, 80131, Napoli, Italy.
Abstract:
The function of p53 protein, also known as "genome guardian", might be impaired by the overexpression of its primary cellular inhibitor, the murine double minute 2 protein (MDM2). However, the recent finding that MDM2-selective inhibitors induce high levels of its homologue MDM4, prompt us to identify, through a receptor-based virtual screening on an in house database, dual MDM2/MDM4 binders. Compound 1 turned out to possess an IC50 of 93.7 and of 4.6 nM on MDM2 and MDM4, respectively. A series of compounds were synthesized to optimize its activity on MDM2. As a result, compound 12 showed low nanomolar IC50 for both targets. NMR studies confirmed the pocket of binding of 12 as predicted by the Glide docking software. Notably, 12 was able to cause concentration-dependent inhibition of cell proliferation, yielding an IC50 value of 356 ± 21 nM in neuroblastoma SHSY5Y cells and proved even to efficiently block cancer stem cell growth.
Insights
Researchers developed dual MDM2/MDM4 inhibitors to overcome resistance to cancer therapies. Compound 12 effectively inhibits cancer cell proliferation and cancer stem cell growth, offering a promising new therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Drug Discovery
Background:
- The p53 protein, or "genome guardian", is crucial for preventing cancer.
- Overexpression of MDM2 (murine double minute 2) protein inhibits p53.
- MDM2 inhibitors can increase MDM4 levels, leading to therapeutic resistance.
Purpose of the Study:
- To identify novel dual inhibitors targeting both MDM2 and MDM4.
- To develop compounds that overcome resistance associated with MDM2-selective inhibitors.
Main Methods:
- Receptor-based virtual screening of an in-house compound database.
- Synthesis and optimization of identified lead compounds.
- In vitro assays to determine inhibitory concentrations (IC50) against MDM2 and MDM4.
- Nuclear Magnetic Resonance (NMR) studies and Glide docking for binding confirmation.
- Cell proliferation assays in neuroblastoma cell lines and cancer stem cells.
Main Results:
- Virtual screening identified initial dual MDM2/MDM4 binders, with Compound 1 showing nanomolar activity.
- Optimization led to Compound 12, exhibiting low nanomolar IC50 values for both MDM2 and MDM4.
- NMR and docking studies confirmed Compound 12's binding site.
- Compound 12 demonstrated concentration-dependent inhibition of neuroblastoma cell proliferation (IC50 = 356 ± 21 nM).
- Compound 12 effectively inhibited cancer stem cell growth.
Conclusions:
- Dual MDM2/MDM4 inhibition is a viable strategy to overcome resistance to MDM2-targeted therapies.
- Compound 12 represents a potent dual inhibitor with significant anti-proliferative and anti-cancer stem cell activity.
- This research offers a promising new avenue for developing novel cancer therapeutics.
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