A library of N-benzylbenzamide, N-phenethylbenzamide and N-benzyloxybenzamide derivatives were designed, synthesized and evaluated as amyloid beta (Aβ42) aggregation inhibitors. These compounds were designed by replacing the α,β-unsaturated linker region of chalcone with an amide bioisostere. The Aβ42 aggregation inhibition properties of these 27 benzamide derivatives were evaluated by the thioflavin T (ThT)-based fluorescence aggregation kinetics assay, transmission electron microscopy (TEM) studies, Aβ42-induced cytotoxicity assay in mouse hippocampal neuronal HT22 cell lines, fluorescence live cell imaging, and computational modelling studies using a pentamer model of Aβ42. These studies led to the identification of N-benzylbenzamides 3a and 3f, N-phenethylbenzamide 5a and N-benzyloxybenzamide 7a as promising compounds that were able to exhibit anti-aggregation properties in the ThT-based fluorescence experiments, TEM studies and more significantly were able to rescue the hippocampal neuronal HT22 cells from Aβ42-induced cytotoxicity (91–96% cell viability at 25 µM). These results demonstrate the usefulness of these benzamide-based templates in the design and development of novel small molecules as chemical tools and therapeutics to study and treat Alzheimer’s disease.

设计、合成了 N-苄基苯甲酰胺、N-苯乙基苯甲酰胺和 N-苄氧基苯甲酰胺衍生物库，并对其作为淀粉样蛋白 β (Aβ42) 聚集抑制剂进行了评估。这些化合物是通过用酰胺生物等排体取代查耳酮的 α,β-不饱和连接区域而设计的。通过基于硫代黄素 T (ThT) 的荧光聚集动力学测定、透射电子显微镜 (TEM) 研究、小鼠海马神经元 HT22 细胞系中 Aβ42 诱导的细胞毒性测定、荧光活细胞评估了这 27 种苯甲酰胺衍生物的 Aβ42 聚集抑制特性。使用 Aβ42 五聚体模型进行成像和计算建模研究。这些研究确定了 N-苄基苯甲酰胺 3a 和 3f、N-苯乙基苯甲酰胺 5a 和 N-苄氧基苯甲酰胺 7a 是有前景的化合物，能够在基于 ThT 的荧光实验、TEM 研究中表现出抗聚集特性，更重要的是能够拯救海马神经元 HT22 细胞免受 Aβ42 诱导的细胞毒性（25 µM 时细胞活力为 91-96%）。这些结果证明了这些基于苯甲酰胺的模板在设计和开发新型小分子作为研究和治疗阿尔茨海默病的化学工具和疗法中的有用性。