NLRP3-IN-33 (Compound 12o) is an inhibitor of AChE and BChE capable of crossing the blood-brain barrier, with IC50 values for hAChE and hBChE at 1.02 μM and 7.03 μM, respectively. NLRP3-IN-33 exhibits antioxidant, anti-inflammatory, and metal chelating activities, making it a candidate for Alzheimer's disease (AD) research.

NLRP3-IN-33 (12o), at concentrations ranging from 1 to 30 μM for 24 hours, exhibits no significant cytotoxicity in PC-12 cells. This compound demonstrates antioxidant properties by effectively inhibiting free radical production, with an IC₅₀ value of 6.19 μM. When used at 12o (1-20 μM; 24 h) in PC-1 cells, NLRP3-IN-33 significantly alleviates H₂O₂ (600 μM; 24 h) induced oxidative stress, offering neuroprotective effects. Additionally, at doses of 1-20 μM for 24 hours in PC-1 cells, it suppresses the activation of the NLRP3 inflammasome and reduces mitochondrial ROS generation and MMP damage induced by LPS (1 μg/mL) and ATP (5 mM) in HMC-3 cells.

NLRP3-IN-33 (12o) at low concentrations (0.05 mg/mL) effectively reduces mitochondrial and cellular oxidative stress in a Drosophila AD model. Additionally, LRP3-IN-33 (5 mg/kg; intraperitoneal injection; once daily for 22 consecutive days) improves memory and cognitive impairments in a scopolamine-induced (1.4 mg/kg; intraperitoneal injection; once daily for 5 consecutive days) AD mouse model.