Pyrazolamide derivatives inhibit α-Synuclein aggregation, disaggregate preformed fibers, and reduce inclusion...

α-Syn fibers, the primary cause and central element of Lewy bodies (LB), play a pivotal role in the development of Parkinson's disease (PD). This research aims to identify more potent inhibitors of α-Syn aggregation. A series of N-aryl-3-aryl-pyrazole-5-carboxamide derivatives were designed and synthesized for this purpose. Among them, four candidate compounds, combining pyrazole and polyphenol blocks, were identified through screening, demonstrating good inhibitory effects with IC₅₀ values in the low micromolar range (1.25-4.29 μM). Two candidates exhibited high permeability through the blood-brain barrier. Mechanistic studies using various methods revealed that the candidates preferentially bind to the aggregation-prone domains-proNAC or NAC domains of α-Syn. This binding hinders the conformational transition from random coil/α-helix to β-sheet, preserving α-Syn proteostasis. As a result, it interferes with α-Syn nuclei formation, prolongs the lag phase, decelerates the elongation phase, and ultimately impedes the formation of α-Syn fibrils. Additionally, the candidates demonstrated promising results in the disaggregation of preformed α-Syn fibers, potentially by binding to specific sites near the β-sheet domain within fibers. This reduces fiber stability, causing rapid collapse and yielding smaller aggregates and monomers. Crucially, the candidate compounds exhibited significant inhibitory efficacy against α-Syn aggregation within nerve cells with low cytotoxicity. This resulted in a notable inhibition of the formation of LB-like α-Syn inclusions. These compounds show considerable promise as potential therapeutic agents for the prevention and treatment of PD.