Multiple sclerosis (MS) is an inflammatory and demyelinating disease of the central nervous system (CNS) that remains incurable. Herein, we demonstrated that ilepcimide (Antiepilepsirine), an antiepileptic drug used for decades, protects mice from experimental autoimmune encephalomyelitis (EAE), a mouse model of MS. Our studies found that ilepcimide treatment effectively ameliorates demyelination, blood-brain barrier leakage and infiltration of CD4⁺ and CD8⁺ T cells in EAE mice. On the one hand, ilepcimide can inhibit dihydroorotate dehydrogenase (DHODH), an important therapeutic target for MS. Computer molecular docking, thermal shift and fluorescence quenching assay demonstrated the directly interaction between ilepcimide and DHODH. Accordingly, ilepcimide observably repressed T cell proliferation in mixed lymphocyte reaction (MLR) assay and concanavalin A (Con-A) model in a DHODH-dependent manner. On the other hand, ilepcimide exhibited neuroprotective effect possibly through activating NRF2 antioxidant pathway in mouse neural crest-derived Neuro2a cells. Collectively, our findings have revealed the therapeutic potential of ilepcimide in EAE mouse model via restricting inflammatory response and oxidative stress, offering a potential opportunity for repurposing existing drug ilepcimide for MS therapy.

多发性硬化症 (MS) 是一种中枢神经系统 (CNS) 炎症和脱髓鞘疾病，至今仍无法治愈。在此，我们证明了 ilepcimide (Antiepilepsirine) 是一种使用了几十年的抗癫痫药物，它可以保护小鼠免受实验性自身免疫性脑脊髓炎 (EAE)（一种 MS 小鼠模型）的侵害。我们的研究发现，ilepcimide 治疗可有效改善脱髓鞘、血脑屏障渗漏和 CD4 ⁺和 CD8 ^+浸润EAE 小鼠中的 T 细胞。一方面，ilepcimide 可以抑制二氢乳清酸脱氢酶 (DHODH)，这是 MS 的重要治疗靶点。计算机分子对接、热位移和荧光猝灭测定证明了 ilepcimide 和 DHODH 之间的直接相互作用。因此，ilepcimide 在混合淋巴细胞反应 (MLR) 测定和伴刀豆球蛋白 A (Con-A) 模型中以 DHODH 依赖性方式可观察到抑制 T 细胞增殖。另一方面，ilepcimide 可能通过激活小鼠神经嵴来源的 Neuro2a 细胞中的 NRF2 抗氧化途径表现出神经保护作用。总的来说，我们的研究结果揭示了 ilepcimide 在 EAE 小鼠模型中的治疗潜力限制炎症反应和氧化应激，为将现有药物 ilepcimide 重新用于 MS 治疗提供了潜在机会。