Amyotrophic lateral sclerosis (ALS) is a devastating motor disease with limited treatment options. A domestic fungal extract library was screened using three assays related to the pathophysiology of ALS with the aim of developing a novel ALS drug. 2(3H)-dihydrofuranolactones 1 and 2, and five known compounds 3–7 were isolated from Pleosporales sp. NUH322 culture media, and their protective activity against the excitotoxicity of β-N-oxalyl-L-α,β-diaminopropionic acid (ODAP), an α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamatergic agonist, was evaluated under low mitochondrial glutathione levels induced by ethacrynic acid (EA) and low sulfur amino acids using our developed ODAP-EA assay. Additional assays evaluated the recovery from cytotoxicity caused by transfected SOD1-G93A, an ALS-causal gene, and the inhibitory effect against reactive oxygen species (ROS) elevation. The structures of 1 and 2 were elucidated using various spectroscopic methods. We synthesized 1 from D-ribose, and confirmed the absolute structure. Isolated and synthesized 1 displayed higher ODAP-EA activities than the extract and represented its activity. Furthermore, 1 exhibited protective activity against SOD1-G93A-induced toxicity. An ALS mouse model, SOD1-G93A, of both sexes, was treated orally with 1 at pre- and post-symptomatic stages. The latter treatment significantly extended their lifespan (p = 0.03) and delayed motor deterioration (p = 0.001–0.01). Our result suggests that 1 is a promising lead compound for the development of ALS drugs with a new spectrum of action targeting both SOD1-G93A proteopathy and excitotoxicity through its action on the AMPA-type glutamatergic receptor.

Graphical abstract

中文翻译：

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来自 Pleosporales sp. 的 2(3H)-二氢呋喃内酯代谢物。NUH322作为抗肌萎缩侧索硬化症药物

肌萎缩侧索硬化症 (ALS) 是一种毁灭性的运动疾病，治疗选择有限。采用三种与 ALS 病理生理学相关的检测方法筛选国内真菌提取物文库，旨在开发一种新型 ALS 药物。从 Pleosporales sp. 中分离出2(3 H )-二氢呋喃内酯1和2以及五种已知化合物 3 – 7 。NUH322 培养基及其对 β- N-草酰-L-α,β-二氨基丙酸 (ODAP)、α-氨基-3-羟基-5-甲基-4-异恶唑丙酸 (AMPA) 兴奋性毒性的保护活性-型谷氨酸激动剂，使用我们开发的 ODAP-EA 测定法在利尿酸 (EA) 和低硫氨基酸诱导的低线粒体谷胱甘肽水平下进行评估。其他测定评估了转染的 SOD1-G93A（一种 ALS 致病基因）引起的细胞毒性的恢复，以及对活性氧 (ROS) 升高的抑制作用。 使用各种光谱方法阐明了1和2的结构 。我们 从D-核糖合成了1 ，并确认了绝对结构。分离和合成的 1 显示出比提取物更高的ODAP-EA活性并代表其活性。此外，  1 对 SOD1-G93A 诱导的毒性表现出保护活性。一种男女 ALS 小鼠模型 SOD1-G93A，在症状出现前和症状出现后阶段均口服 1  。后一种治疗显着延长了他们的寿命（p = 0.03）并延迟了运动退化（p = 0.001–0.01）。我们的结果表明， 1 是一种很有前景的先导化合物，可用于开发 ALS 药物，具有针对 SOD1-G93A 蛋白病和通过作用于 AMPA 型谷氨酸受体而产生的兴奋性毒性的新作用谱。

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