Özgün Caspase-1 ve IL1R inhibitörlerinin tanımlanması ve karakterize edilmesi

Autoinflammatory diseases occur when the body's innate immune system is abnormally activated. There are more than eighty types of autoinflammatory diseases which affect >5% of the population in Western countries. Elevated levels of Interleukin-1 (IL1) cytokines, especially IL1β and IL1α, are observed in many types of autoinflammatory diseases including Behcet's Disease and Familial Mediterranean Fever (FMF). IL1β binds Interleukin-1 Receptor (IL1R) and initiates NLRP3 (cryopyrin) inflammasome activation. Aberrant levels of IL1R signaling and inflammasome activation promote autoinflammatory diseases. With such a critical role there are several treatment options to reduce inflammasome activation including: caspase-1 inhibitors which are not FDA proved, IL1R antagonists and neutralizing α-IL1β antibodies. Because the known therapies are protein-based, they are costly and require daily injections. However there are no identified IL1R small molecule inhibitors which can be less costly and overcome the injection barriers. Therefore the goal of this thesis was to identify and characterize novel small molecule inflammasome inhibitors. For this, we utilized in silico modeling in combination with drug repurposing to develop small molecule inhibitors which target IL1R signaling pathway. We screened >300 indole derivatives, and 1280 FDA-approved drugs from the Prestwick Library in a cell-based assay. From this we identified several compounds with mid-nanomolar IC50 and no obvious cellular toxicity. Further studies suggest that these are reversible inhibitors. To better understand the mechanism of inhibition, we developed a docking model to identify possible binding sites of small molecules and performed mutagenesis analysis with the predicted binding pocket. Our experimental data suggests that our small molecule inhibitors possibly inhibit some other protein/s in the down signaling mechanism of in the IL1R pathway. For future studies, we are going to try to iv characterize this/these protein/s. In future experiments, animal models will be utilized for the biological analysis of our small molecule inhibitors.