Lee Yeongyeong, Han Sukmin, Lee Jeongmi, Cho Yongeun, Kim Jun-Sik, Jeon Yeji, Cho Heewon, Yoo Heejin, Byun Yujung, Kim Tai Kyoung, Hong Ju-Mi, Kim Hyunwook, Park Sang Yoon, Yim Joung Han, Kim Sung Hyun, Jo Dong-Gyu
School of Pharmacy, Sungkyunkwan University, Suwon, 16419, Republic of Korea.
Department of Neuroscience, Graduate School, Kyung Hee University, Seoul, 02447, Republic of Korea.
Arch Pharm Res. 2025 Aug 6. doi: 10.1007/s12272-025-01562-0.
Alzheimer's disease (AD) is characterized by progressive cognitive decline, amyloid plaque accumulation, synaptic dysfunction, and neuroinflammation. This study reports the therapeutic potential of (S)-4-amino-5,5-difluoro-N'-methyl-N'-phenylpentanehydrazide hydrochloride (RA-058HM), a novel compound, in ameliorating these pathological features of AD in the 5xFAD mouse model. RA-058HM was administered orally for 8 weeks, and its multi-target effects - including relief from neuroinflammation, normalization of synaptic transmission, reduction of amyloidogenesis (plaque and soluble oligomers, as well as BACE1 levels), and rescue of cognitive function-were evaluated. To our knowledge, RA-058HM is the first compound to demonstrate simultaneous modulation of these key pathways in the 5xFAD model, highlighting its potential as a comprehensive disease-modifying therapy for AD. Behavioural tests revealed marked improvements in spatial and recognition memory in RA-058HM-treated 5xFAD mice, suggesting a reversal of cognitive deficits. At the molecular level, RA-058HM treatment reduced amyloidogenesis, as evidenced by decreased levels of amyloid precursor protein (APP) and β-secretase (BACE1) in the hippocampus, accompanied by reduced plaque formation, as detected by Thioflavin-S staining. Furthermore, synaptic transmission was restored to near-normal levels in RA-058HM-treated neurons, indicating that RA-058HM effectively rescues synaptic deficits without altering synaptic protein levels of PSD95 and synaptophysin. In addition, treatment of RA-058HM downregulated hippocampal levels of the NLRP3 inflammasome, TNF-α, and GFAP, suggesting a decrease in neuroinflammatory signaling and a modulation of glial activity. Restoration of mitochondrial motility in hippocampal neurons further suggests that RA-058HM may improve cellular energy dynamics. Collectively, these findings indicate that RA-058HM has multifaceted effects on AD pathology, targeting amyloid accumulation, synaptic transmission, neuroinflammation, and mitochondrial function. This study highlights RA-058HM as a promising candidate for AD therapy and underscores the potential of multi-targeted approaches in addressing the complex mechanisms underlying AD progression.
阿尔茨海默病（AD）的特征是进行性认知衰退、淀粉样斑块积累、突触功能障碍和神经炎症。本研究报告了一种新型化合物（S）-4-氨基-5,5-二氟-N'-甲基-N'-苯基戊烷酰肼盐酸盐（RA-058HM）在改善5xFAD小鼠模型中AD的这些病理特征方面的治疗潜力。RA-058HM口服给药8周，并评估其多靶点效应，包括减轻神经炎症、使突触传递正常化、减少淀粉样蛋白生成（斑块和可溶性寡聚体以及β-分泌酶1水平）以及挽救认知功能。据我们所知，RA-058HM是第一个在5xFAD模型中证明能同时调节这些关键途径的化合物，突出了其作为AD综合疾病修饰疗法的潜力。行为测试显示，经RA-058HM治疗的5xFAD小鼠在空间和识别记忆方面有显著改善，表明认知缺陷得到了逆转。在分子水平上，RA-058HM治疗减少了淀粉样蛋白生成，海马体中淀粉样前体蛋白（APP）和β-分泌酶（BACE1）水平降低证明了这一点，硫黄素-S染色检测显示斑块形成减少。此外，经RA-058HM治疗的神经元中突触传递恢复到接近正常水平，表明RA-058HM有效地挽救了突触缺陷，而不会改变PSD95和突触素的突触蛋白水平。此外，RA-058HM治疗下调了海马体中NLRP3炎性小体、肿瘤坏死因子-α和胶质纤维酸性蛋白的水平，表明神经炎症信号减少和神经胶质细胞活性得到调节。海马体神经元中线粒体运动性的恢复进一步表明，RA-058HM可能改善细胞能量动态。总体而言，这些发现表明RA-058HM对AD病理具有多方面的影响，靶向淀粉样蛋白积累、突触传递、神经炎症和线粒体功能。本研究突出了RA-058HM作为AD治疗有前景的候选药物，并强调了多靶点方法在解决AD进展潜在复杂机制方面的潜力。
