Identification of the first non-peptidic small molecule inhibitor of the c-Abl/14-3-3 protein–protein interactions able to drive sensitive and Imatinib-resistant leukemia cells to apoptosis

Corradi, Valentina; Mancini, Manuela; Manetti, Fabrizio; Petta, Sara; Santucci, Maria Alessandra; Botta, Maurizio

doi:10.1016/j.bmcl.2010.08.019

Cited by 71 publications

(69 citation statements)

References 16 publications

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“…2). For example, the rmsd of the Ca atoms of MLF1 (31)(32)(33)(34) and histone H3(7-10) is 0.28 Å whereas the rmsd of the four corresponding residues after the phosphorylated serine (MLF1 (35)(36)(37)(38) and histone H3 (11)(12)(13)(14)) is 1.49 Å .…”

Section: Resultsmentioning

confidence: 99%

“…In the sample cell, a solution of 50 lM 14-3-3 protein was placed and titrated stepwise with 6 lL aliquots of a 500 lM solution of MLF1 (29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)pSer34. The association constant K a (K a = 1 ⁄ K D ), molar binding stoichiometry (N) and molar binding enthalpy (DHº) were determined by fitting the binding isotherm by a single binding site model.…”

Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

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Structural insights of the MLF1/14‐3‐3 interaction

et al. 2012

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Myeloid leukaemia factor 1 (MLF1) binds to 14-3-3 adapter proteins by a sequence surrounding Ser34 with the functional consequences of this interaction largely unknown. We present here the high-resolution crystal structure of this binding motif pSer34] in complex with 14-3-3e and analyse the interaction with isothermal titration calorimetry. Fragment-based ligand discovery employing crystals of the binary 14-3-3e ⁄ MLF1(29-42)pSer34 complex was used to identify a molecule that binds to the interface rim of the two proteins, potentially representing the starting point for the development of a small molecule that stabilizes the MLF1 ⁄ 14-3-3 protein-protein interaction. Such a compound might be used as a chemical biology tool to further analyse the 14-3-3 ⁄ MLF1 interaction without the use of genetic methods. Structured digital abstract l 14-3-3 epsilon and MLF1 bind by x-ray crystallography (View interaction) l 14-3-3 epsilon and MLF1 bind by isothermal titration calorimetry (View Interaction: 1, 2)

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Section: Resultsmentioning

confidence: 99%

Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

Structural insights of the MLF1/14‐3‐3 interaction

et al. 2012

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“…This resulted in the identification of BV02 (12) as a lead inhibitor of the interaction between 14-3-3σ and cAbl in chronic myelogenous leukemia (CML) ( Figure 9C). 124 Indeed, 12 was able to inhibit 14-3-3/c-Abl interaction and promote c-Abl nuclear translocation at low micromolar concentration in Ba/F3 cells expressing the wild-type Bcr-Abl as well as its Imatinib-resistant T315I mutation. 125 Accordingly, 12 represented a useful starting point for the development of an alternative treatment of CML, particularly for the Imatinibresistant forms.…”

Section: Journal Of Medicinal Chemistrymentioning

confidence: 96%

“…127 This discovery was facilitated by in silico docking of a virtual library of 12 and 10 analogues to a 14-3-3σ crystal structure using a well-established computational protocol. 124 The most promising molecules were synthesized and submitted to biological tests. Most notably, molecule 13 was found to promote c-Abl nuclear translocation at 25 μM and has been shown to decrease the IC 50 of doxorubicin by increasing its accumulation in MDR cancer cells at 10 μM concentration.…”

Section: Journal Of Medicinal Chemistrymentioning

confidence: 99%

Modulators of Protein–Protein Interactions

et al. 2014

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Direct interactions between proteins are essential for the regulation of their functions in biological pathways. Targeting the complex network of protein−protein interactions (PPIs) has now been widely recognized as an attractive means to therapeutically intervene in disease states. Even though this is a challenging endeavor and PPIs have long been regarded as "undruggable" targets, the last two decades have seen an increasing number of successful examples of PPI modulators, resulting in growing interest in this field. PPI modulation requires novel approaches and the integrated efforts of multiple disciplines to be a fruitful strategy. This perspective focuses on the hub-protein 14-3-3, which has several hundred identified protein interaction partners, and is therefore involved in a wide range of cellular processes and diseases. Here, we aim to provide an integrated overview of the approaches explored for the modulation of 14-3-3 PPIs and review the examples resulting from these efforts in both inhibiting and stabilizing specific 14-3-3 protein complexes by small molecules, peptide mimetics, and natural products.

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“…19 We previously performed an HTS of the LOPAC library using an FP-based assay for the interaction of 14-3-3g and Raf-1 protein, an interaction critical for mitogenic signal transduction, 16 and identified a small molecule compound, FOBSIN101, as a 14-3-3 protein inhibitor. 20 In addition to FOBOSIN101, several other small-molecule 14-3-3 inhibitors have been identified through computational-based virtual screening 21 and fragment-based combinatorial small-molecule microarray. 22,23 However, none of these reported compounds showed both high potency and on-target effect in any animal model systems.…”

Section: Introductionmentioning

confidence: 99%

A Time-Resolved Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of 14-3-3 Protein–Protein Interaction Inhibitors

Liu

et al. 2013

ASSAY and Drug Development Technologies

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Protein-protein interaction networks mediate diverse biological processes by regulating various signaling hubs and clusters. 14-3-3 proteins, a family of phosphoserine/threonine-binding molecules, serve as major interaction hubs in eukaryotic cells and have emerged as promising therapeutic targets for various human diseases. In order to identify chemical probes for mechanistic studies and for potential therapeutic development, we have developed highly sensitive bioassays to monitor the interaction of 14-3-3 with a client protein. In this study, we describe a homogenous time-resolved fluorescence resonance energy transfer (TR-FRET) assay to detect the interaction of 14-3-3 with Bad, a proapoptotic member of the Bcl-2 family. Through a series of titration studies in which europium-labeled 14-3-3 serves as an FRET donor and a Dy647-labeled phosphorylated Bad, the peptide acts as an FRET acceptor, we have achieved a robust TR-FRET assay that is suitable for high-throughput screening (HTS) with an excellent signal-to-background ratio of >20 and Z 0 values >0.7. This assay was further miniaturized to a 1,536-well format for ultra-HTS (uHTS), and exhibited a similar robust performance. The utility and performance of the assay for uHTS were validated by (i) known inhibitors, including peptide R18 and small molecule FOBISIN101, and (ii) screening of a 51,200 compound library. This simple and robust assay is generally applicable to detect the interaction of 14-3-3 with other client proteins. It provides a sensitive and easy-to-use tool to facilitate the discovery of 14-3-3 protein inhibitors as well as to study 14-3-3-mediated protein-protein interactions.

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Identification of the first non-peptidic small molecule inhibitor of the c-Abl/14-3-3 protein–protein interactions able to drive sensitive and Imatinib-resistant leukemia cells to apoptosis

Cited by 71 publications

References 16 publications

Structural insights of the MLF1/14‐3‐3 interaction

Structural insights of the MLF1/14‐3‐3 interaction

Modulators of Protein–Protein Interactions

A Time-Resolved Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of 14-3-3 Protein–Protein Interaction Inhibitors

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