Development of NLRP3 inhibitors for intervening in Alzheimer’s disease

Abstract

AbstractBackgroundNLRP3 inflammasome is an essential component of innate immunity, and its dysregulation has been linked to neuroinflammation and the development of neurodegenerative disorders including Alzheimer’s disease. Therefore, NLRP3 inflammasome represents a promising target for effective AD treatment development.MethodBased on our newly identified lead NLRP3 inhibitor, a photoaffinity labeling probe was designed and employed to understand the mechanism of action for inhibitors based on this chemical scaffold using biochemical, biophysical, and molecular biology assays.ResultThe chemical probe can specifically recognize the NLRP3 protein from cell lysates. Photo‐affinity labeling studies using recombinant human NLRP3 protein also showed their interactions, and notably, the mode of action is different than other known NLRP3 inhibitors. Further characterization using FRET confirmed its interaction with the NLRP3 protein. Mass spectrometry analyses of the photo‐affinity labeled NLRP3 protein revealed a novel biding site with the NACHT domain of the NLRP3 protein.ConclusionStudies using a rationally designed photo‐affinity probe based on our lead NLRP3 inhibitor demonstrated that our inhibitors directly bind to the NLRP3 protein via a novel mechanism of action, distinct from other known NLRP3 inhibitors.