Repurposing of cefpodoxime proxetil as potent neuroprotective agent through computational prediction and in vitro validation
Repurposing of cefpodoxime proxetil as potent neuroprotective agent through computational prediction and in vitro validation
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DOI：
10.1080/07391102.2020.1772884
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发表时间：
2020-06-02
影响因子：
4.4
通讯作者：
Cheng, Maosheng
中科院分区：
文献类型：
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作者：
Quan, Jishun;Ma, Chao;Cheng, Maosheng
在最近的报道中，NR2B-NMDA受体拮抗剂因其靶向性强、副作用小而显示出更大的研究价值。2016年，据报道，EVT-101结合在该靶的几乎全新的结合区域。截然不同的结合模式是否可以提高靶向性，减少副作用，值得研究。在前期工作中，我们探索了依芬地尔和EVT-101的结合模式，找到了关键氨基酸，并总结了药效团，希望找到同时针对这两种结合模式的拮抗剂。在这项研究中，我们在FDA批准的药物库中开发了一个可扩展的虚拟筛选工作流程，以基于两个药物载体的组合来识别新的NR2B-NMDAR拮抗剂。头孢泊肟酯(5)被确定为HIT化合物，并首次发现5可能具有神经保护活性作为NR2B-NMDAR拮抗剂。这一结果引起了我们进一步研究的兴趣，通过分子对接研究建立的配体-受体相互作用模型表明，该化合物能够在GluN1和GluN2B的ATD之间的结合空腔完美地融合依芬普地尔和EVT-101的药效团特性。通过100 ns分子动力学模拟和结合自由能计算，验证了分子对接结果的准确性和配体-受体络合物的结合稳定性。然后，对NMDA损伤的SH-SY5Y细胞进行四甲基偶氮唑盐比色试验(49.8%+/-0.1%，5mU M)，并用抑制NMDA诱导的Ca(2+)内流作用的证据来验证计算结果，进一步的研究表明5可以抑制NMDA诱导的NR2B的上调。
In recent reports, NR2B-NMDA receptor antagonists showed more research value because of its strong targeting ability and less side effects potential. In 2016, EVT-101 was reported to bind in an almost entirely new binding region of this target. Whether strikingly different binding modes can improve targeting and reduce side effects is worth studying. In our preliminary work, we explored the binding patterns of ifenprodil and EVT-101, found the key amino acids and summarized the pharmacophores, hoping to find such antagonists that target the two binding modes simultaneously. In this study, we developed a scalable virtual screening workflow in the FDA-approved drugs library to identify novel NR2B-NMDAR antagonists based on the combination of two pharmacophores. Cefpodoxime proxetil (5) was identified as the hit compound, and it was found for the first time that5might have neuroprotective activity as a NR2B-NMDAR antagonist. This result interested us to make further study, the ligand-receptor interactions modeled by molecular docking studies showed that the compound could perfectly merge both the pharmacophore characteristics of ifenprodil and EVT-101 at the binding cavity between the ATDs of GluN1 and GluN2B. The accuracy of molecular docking results and binding stability of ligand-receptor complexes were validated through 100 ns molecular dynamics simulation and binding free energy calculation. Afterwards, MTT assay (49.8%+/- 0.1%, 5 mu M) on NMDA injured SH-SY5Y cells and evidence of the effect on attenuating Ca(2+)influx induced by NMDA were applied to validate the computational results, further investigation showed that5could suppress the NR2B upregulation induced by NMDA.