Fig. 4

Screening strategy to identify compounds that inhibit α-synuclein oligomers and α-synuclein-mediated cytotoxicity. An AI-driven in silico screen was first performed with IBM Watson for Drug Discovery Predictive Analytics to predict compounds with a high likelihood of inhibiting aggregation of α-synuclein into oligomers. Next, highly ranked drugs were tested for their ability to reduce α-synuclein oligomer levels in vitro using a Gaussia princeps luciferase protein-fragment complementation cell assay. Positive hits were then assessed in vivo by measuring their effects on the motor impairment of C. elegans expressing α-synuclein. Last, screen hits were validated in mammalian in vitro and in vivo models. Specifically, drugs that reduced the coiling of C. elegans were examined for their ability to reduce α-synuclein oligomers measured by YFP protein-fragment complementation in rat primary cortical neurons, and our top candidate was tested in an AAV-based rat model of α-synuclein-mediated dopaminergic neurodegeneration