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 anti-helminthic 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.

中文翻译：

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在计算机上进行分子靶预测，发现甲苯苯达唑为有效的MAPK14抑制剂

多元药理学的概念涉及药物分子与多个分子靶标的相互作用。它为重新利用已经批准的药物针对人类疾病的关键因素提供了独特的机会。在本文中，我们使用了计算机模拟目标预测算法来研究甲苯咪唑（一种抗蠕虫药）的作用机理，该药物目前被重新用于治疗脑肿瘤。首先，我们证实了甲苯咪唑降低了胶质母细胞瘤细胞的体外生存能力。我们的计算机模拟方法揭示了21种可能的甲苯达唑分子靶标，包括与正常脑组织相比在胶质母细胞瘤的基因水平上显着上调的12种蛋白质。对参与癌症生物学的三种主要激酶进行了验证实验：ABL1，MAPK1 / ERK2和MAPK14 /p38α。甲苯达唑可以剂量依赖性方式体外抑制这些激酶的活性，对MAPK14具有很高的效力。它与MAPK14的直接结合在体外得到进一步验证，并在活胶质母细胞瘤细胞中证实了MAPK14激酶活性的抑制作用。与生物物理数据一致，分子模型表明甲苯达唑能够与MAPK14的催化位点结合。最后，基因沉默表明MAPK14参与了胶质母细胞瘤肿瘤球体的生长和对甲苯达唑治疗的反应。因此，这项研究强调了MAPK14在甲苯咪唑的抗癌作用机制中的作用，并为脑肿瘤中MAPK14的药理靶向提供了进一步的理论依据。这也为开发新型MAPK14 /p38α抑制剂以治疗人类疾病开辟了新途径。