In silico molecular target prediction unveils mebendazole as a potent MAPK14 inhibitor

Abstract

The concept of polypharmacology involves the interaction of drug molecules with multiple molecular targets. It provides a unique opportunity for the repurposing of already-approved drugs to target key factors involved in human diseases. Herein, we used an in silico target prediction algorithm to investigate the mechanism of action of mebendazole, an antihelminthic drug, currently repurposed in the treatment of brain tumors. First, we confirmed that mebendazole decreased the viability of glioblastoma cells in vitro. Our in silico approach unveiled 21 putative molecular targets for mebendazole, including 12 proteins significantly up-regulated at the gene level in glioblastoma as compared to normal brain tissue. Validation experiments were performed on three major kinases involved in cancer biology: ABL1, MAPK1/ERK2 and MAPK14/p38α. Mebendazole could inhibit the activity of these kinases in vitro in a dose-dependent manner, with a high potency against MAPK14. Its direct binding to MAPK14 was further validated in vitro and inhibition of MAPK14 kinase activity was confirmed in live glioblastoma cells. Consistent with biophysical data, molecular modeling suggested that mebendazole was able to bind to the catalytic site of MAPK14. Finally, gene silencing demonstrated that MAPK14 is involved in glioblastoma tumor spheroid growth and response to mebendazole treatment. This study thus highlighted the role of MAPK14 in the anticancer mechanism of action of mebendazole and provides further rationale for the pharmacological targeting of MAPK14 in brain tumors. It also opens new avenues for the development of novel MAPK14/p38α inhibitors to treat human diseases. Significance StatementThis study provides a framework to investigate drug polypharmacology by rapidly identifying novel molecular targets of already-approved drugs. It unveils a new mechanism involved in the anticancer activity of anti-helminthic drug, mebendazole, which is currently being repurposed for the treatment of brain tumors. By helping to decipher the mechanism(s) of action of repurposed drugs in their new indications, this approach could contribute to the development of safer and more effective therapeutic strategies in oncology and beyond.

Identification and Validation of Carbonic Anhydrase II as the First Target of the Anti-Inflammatory Drug Actarit

“…Studies including reviews of computational methods for target prediction have been published [ 3 , 4 ]. These methods have shown their utility for tackling these problems, both retrospectively [ 5 , 6 ] and prospectively [ 7 , 8 , 9 ]. An early example of these methods is the Similarity Ensemble Approach (SEA), which constructs a statistical model for each considered target and uses the ensemble of models to predict which targets interact with the investigated molecule.…”

“…We developed MolTarPred [ 4 ], a chemical-similarity-based method able to predict more than 4500 protein targets, and made it freely available as a webserver [ 10 ]. This version of MolTarPred has recently predicted new targets for mebendazole and nocodazole [ 9 ]. These predicted targets were confirmed in vitro: mebendazole inhibit MAPK14 with an IC 50 of 104 nM and nocodazole inhibit ABL1 with an IC 50 of 78 nM [ 9 ].…”

“…This version of MolTarPred has recently predicted new targets for mebendazole and nocodazole [ 9 ]. These predicted targets were confirmed in vitro: mebendazole inhibit MAPK14 with an IC 50 of 104 nM and nocodazole inhibit ABL1 with an IC 50 of 78 nM [ 9 ]. Although several targets were already known for these two anti-helminthic drugs, the discovery of their potent activity against these cancer targets led to a better explanation of their anti-glioma properties [ 9 ].…”

Beta-blockers disrupt mitochondrial bioenergetics and increase radiotherapy efficacy independently of beta-adrenergic receptors in medulloblastoma

“…IC 50 values were determined as previously described. 28 For IncuCyte experiments (RRID:SCR_019874), SHH MB tumour cells were plated in 96-well plates (5,000 cells/well for murine SHH-MB and 7,500 cells/well for ICN-MB-PDX-12), pre-coated with poly-D-lysine (EMD Millipore, ref. A-003-E) and Matrigel (BD Biosciences,ref.…”

Mebendazole’s Conformational Space and its Predicted Binding to Human Heat-Shock Protein 90

“…However, few computational predictions have been made for mebendazole's binding to various proteins. [65][66][67][68] Recently, we experimentally showed that mebendazole is a potent Hedgehog signaling inhibitor that acts by inducing proteasomal degradation of glioma-associated oncogene (GLI) transcription factors. 57 GLI transcription factors drive a genetic program being fundamental for the pathophysiology of leukemia initiating cells that are responsible for therapy failure and tumor relapse due to their chemotherapy resistance.…”