Glutamate toxicity is a pathomechanism that contributes to neuronal cell death in a wide range of acute and chronic neurodegenerative and neuroinflammatory diseases. Activation of the N-methyl-D-aspartate (NMDA)-type glutamate receptor and breakdown of the mitochondrial membrane potential are key events during glutamate toxicity. Due to its manifold functions in nervous system physiology, however, the NMDA receptor is not well suited as a drug target. To identify novel compounds that act downstream of toxic NMDA receptor signaling and can protect mitochondria from glutamate toxicity, we developed a cell viability screening assay in primary mouse cortical neurons. In a proof-of-principle screen we tested 146 natural products and 424 FDA-approved drugs for their ability to protect neurons against NMDA-induced cell death. We confirmed several known neuroprotective drugs that include Dutasteride, Enalapril, Finasteride, Haloperidol, and Oxybutynin, and we identified neuroprotective properties of Elvitegravir. Using live imaging of tetramethylrhodamine ethyl ester-labelled primary cortical neurons, we found that Elvitegravir, Dutasteride, and Oxybutynin attenuated the NMDA-induced breakdown of the mitochondrial membrane potential. Patch clamp electrophysiological recordings in NMDA receptor-expressing HEK293 cell lines and primary mouse hippocampal neurons revealed that Elvitegravir does not act at the NMDA receptor and does not affect the function of glutamatergic synapses. In summary, we have developed a cost-effective and easy-to-implement screening assay in primary neurons and identified Elvitegravir as a neuro- and mitoprotective drug that acts downstream of the NMDA receptor.

中文翻译：

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初级神经元中基于显微镜的小分子筛选揭示了 FDA 批准的抗病毒药物 Elvitegravir 的神经保护特性。

谷氨酸毒性是一种病理机制，可导致多种急性和慢性神经退行性疾病和神经炎症疾病中的神经元细胞死亡。N-甲基-D-天冬氨酸 (NMDA) 型谷氨酸受体的激活和线粒体膜电位的破坏是谷氨酸毒性期间的关键事件。然而，由于其在神经系统生理学中的多种功能，NMDA 受体不太适合作为药物靶点。为了鉴定在有毒 NMDA 受体信号下游起作用并可以保护线粒体免受谷氨酸毒性的新型化合物，我们在原代小鼠皮层神经元中开发了细胞活力筛选试验。在原理验证筛选中，我们测试了 146 种天然产物和 424 种 FDA 批准的药物，以检测它们保护神经元免受 NMDA 诱导的细胞死亡的能力。我们确认了几种已知的神经保护药物，包括度他雄胺、依那普利、非那雄胺、氟哌啶醇和奥昔布宁，并确定了 Elvitegravir 的神经保护特性。使用四甲基罗丹明乙酯标记的原代皮质神经元的实时成像，我们发现 Elvitegravir、度他雄胺和奥昔布宁减弱了 NMDA 诱导的线粒体膜电位分解。在表达 NMDA 受体的 HEK293 细胞系和原代小鼠海马神经元中的膜片钳电生理记录显示，Elvitegravir 不作用于 NMDA 受体，也不影响谷氨酸能突触的功能。总之，