NLRP3-IN-10 (ZVN26391) is a potent NLRP3 inhibitor. NLRP3-IN-10 inhibits IL-1β release with an IC50 value of 251.1 nM. NLRP3-IN-10 attenuates ASC speck formation, leading to suppress activation of NLRP3 inflammasome.

NLRP3-IN-10 (compound 14c) significantly inhibits NLRP3 inflammasome activation in THP-1 cells by LPS-MSU, demonstrating dose-dependent efficacy at 0.4-6.4 μM over 40 minutes. It is non-toxic to THP-1 cells at 0.1-6.4 μM after 1.5 hours and prevents Nigericin-induced pyroptosis at 0.1 and 0.4 μM. The compound reduces caspase-1 p20 and IL-1β production dose-dependently at 0.1, 0.2, and 0.4 μM, and decreases LPS-induced TNF-α production at 3 and 5 μM. NLRP3-IN-10 also reduces ASC specks formation at 0.2 and 0.8 μM, indicating interruption of ASC oligomerization. Furthermore, it inhibits LPS-induced NLRP3 priming by directly interacting with NLRP3 at 1, 10, and 100 μM, demonstrating potential in regulating both priming and activation steps in the NLRP3 inflammasome pathway.

NLRP3-IN-10 (compound 14c), administered via a single intravenous (i.v.) dose of 10 mg/kg, effectively reduced peritoneal neutrophil influx and IL-1β levels in the spleen in a LPS-primed mouse model with MSU-induced peritonitis. Additionally, oral administration of NLRP3-IN-10 at doses of 10, 30, and 90 mg/kg demonstrated very low systemic exposure (14.6 to 23.53 μg·h/L), limited bioavailability (2.47 to 13.79%), and high plasma clearance rates (2201.58 to 5551.12 L/h/kg), indicating significant challenges in its pharmacokinetic profile when administered orally. Pharmacokinetic data revealed an area under the curve (AUC) ranging from 14.60 to 23.53 μg·h/L, clearance (CL) rates between 2201.58 and 5551.12 L/h/kg, and maximal concentration (Cmax) values from 3.35 to 81.97 μg/L across different administration routes and dosages. The study was conducted on 7-week-old male C57BL/6J mice induced with MSU-induced peritonitis and primed with LPS (1 mg/kg, i.p.), showing a notable reduction in spleen IL-1β release and peritoneal neutrophil influx following a 6-hour treatment with a 10 mg/kg intravenous dose of NLRP3-IN-10, when compared to the control group.