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Insights into the Interaction Mechanism of DTP3 with MKK7 by Using STD-NMR and Computational Approaches
Annamaria Sandomenico1, Lorenzo Di Rienzo2, Luisa Calvanese3
1Institute of Biostructures and Bioimaging (IBB)-CNR, Via Mezzocannone 16, 80134 Naples, Italy.
Abstract:
GADD45β/MKK7 complex is a non-redundant, cancer cell-restricted survival module downstream of the NF-kB survival pathway, and it has a pathogenically critical role in multiple myeloma, an incurable malignancy of plasma cells. The first-in-class GADD45β/MKK7 inhibitor DTP3 effectively kills MM cells expressing its molecular target, both in vitro and in vivo, by inducing MKK7/JNK-dependent apoptosis with no apparent toxicity to normal cells. DTP3 combines favorable drug-like properties, with on-target-specific pharmacology, resulting in a safe and cancer-selective therapeutic effect; however, its mode of action is only partially understood. In this work, we have investigated the molecular determinants underlying the MKK7 interaction with DTP3 by combining computational, NMR, and spectroscopic methods. Data gathered by fluorescence quenching and computational approaches consistently indicate that the N-terminal region of MKK7 is the optimal binding site explored by DTP3. These findings further the understanding of the selective mode of action of GADD45β/MKK7 inhibitors and inform potential mechanisms of drug resistance. Notably, upon validation of the safety and efficacy of DTP3 in human trials, our results could also facilitate the development of novel DTP3-like therapeutics with improved bioavailability or the capacity to bypass drug resistance.
Insights
The GADD45β/MKK7 inhibitor DTP3 targets multiple myeloma cells selectively. This study reveals DTP3 binds the MKK7 N-terminal region, clarifying its cancer-selective action and informing future drug development.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- The GADD45β/MKK7 complex is crucial for cancer cell survival, particularly in multiple myeloma (MM).
- The inhibitor DTP3 demonstrates efficacy against MM cells by inducing apoptosis via MKK7/JNK.
- DTP3 exhibits cancer-selective therapeutic effects with favorable drug-like properties.
Purpose of the Study:
- To elucidate the molecular mechanisms of MKK7 interaction with the inhibitor DTP3.
- To understand the binding site and determinants of DTP3's selective action.
- To inform the development of novel therapeutics and potential resistance strategies.
Main Methods:
- Computational modeling to predict binding interactions.
- Nuclear Magnetic Resonance (NMR) spectroscopy to analyze protein-ligand interactions.
- Spectroscopic methods, including fluorescence quenching, to assess binding.
Main Results:
- Consistent data from fluorescence quenching and computational approaches identified the MKK7 N-terminal region as the primary binding site for DTP3.
- This finding provides a detailed understanding of how DTP3 selectively targets MKK7.
- The results offer insights into potential mechanisms of drug resistance to DTP3.
Conclusions:
- The N-terminal region of MKK7 is the key interaction site for the inhibitor DTP3.
- Understanding this interaction enhances knowledge of GADD45β/MKK7 inhibitor selectivity.
- These findings pave the way for developing improved DTP3-like therapeutics and overcoming drug resistance in multiple myeloma.
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