Pharmacological Analysis of NLRP3 Inflammasome Inhibitor Sodium (1, 2, 3, 5, 6, 7-Hexahydro-s-indacen-4-yl) carbamoyl (1-methyl-1H-pyrazol-4-yl) ([ (2S) -oxolan-2-ylmethyl) sulfamoyl]azanide in Cellular and Mouse Models of Inflammation Provides a Translational Framework

Interleukin (IL) -1β is an apex proinflammatory cytokine produced in response to tissue injury and infection. The output of IL-1β from monocytes and macrophages is regulated not only by transcription and translation but also post-translationally. Release of the active cytokine requires activation of inflammasomes, which couple IL-1β post-translational proteolysis with pyroptosis. Among inflammasome platforms, NOD-like receptor pyrin domain-containing protein 3 (NLRP3) is implicated in the pathogenesis of numerous human disorders in which disease-specific danger-associated molecular patterns (DAMPS) are positioned to drive its activation. As a promising therapeutic target, numerous candidate NLRP3-targeting therapeutics have been described and demonstrated to provide benefits in the context of animal disease models. While showing benefits, published preclinical studies have not explored dose–response relationships within the context of the models. Here, the preclinical pharmacology of a new chemical entity, (1, 2, 3, 5, 6, 7-hexahydro-s-indacen-4-yl) carbamoyl (1-methyl-1H-pyrazol-4-yl) ([ (2S) -oxolan-2-ylmethyl) sulfamoyl]azanide (NT-0249), is detailed, establishing its potency and selectivity as an NLRP3 inhibitor. NT-0249 also is evaluated in two acute in vivo mouse challenge models where pharmacodynamic/pharmacokinetic relationships align well with in vitro blood potency assessments. The therapeutic utility of NT-0249 is established in a mouse model of cryopyrin-associated periodic syndrome (CAPS). In this model, mice express a human gain-of-function NLRP3 allele and develop chronic and progressive IL-1β-dependent autoinflammatory disease. NT-0249 dose-dependently reduced multiple inflammatory biomarkers in this model. Significantly, NT-0249 decreased mature IL-1β levels in tissue homogenates, confirming in vivo target engagement. Our findings highlight not only the pharmacological attributes of NT-0249 but also provide insight into the extent of target suppression that will be required to achieve clinical benefit.