Overcoming Preclinical Safety Obstacles to Discover (S)-N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-6-(methylamino)-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine-3-sulfonamide (GDC-2394): A Potent and Selective NLRP3 Inhibitor

Abstract

Inappropriate activation of the NLRP3 inflammasome has been implicated in multiple inflammatory and autoimmune diseases. Herein, we aimed to develop novel NLRP3 inhibitors that could minimize the risk of drug-induced liver injury. Lipophilic ligand efficiency was used as a guiding metric to identify a series of 6,7-dihydro-5H-pyrazolo­[5,1-b]­[1,3]­oxazinesulfonylureas. A leading compound from this series was advanced into safety studies in cynomolgus monkeys, and renal toxicity, due to compound precipitation, was observed. To overcome this obstacle, we focused on improving the solubility of our compounds, specifically by introducing basic amine substituents into the scaffold. This led to the identification of GDC-2394, a potent and selective NLRP3 inhibitor, with an in vitro and in vivo safety profile suitable for advancement into human clinical trials.

First‐in‐human phase 1 trial evaluating safety, pharmacokinetics, and pharmacodynamics of NLRP3 inflammasome inhibitor, GDC‐2394, in healthy volunteers

“…Of note, the 150‐mg starting dose in this study was anticipated to be subtherapeutic based on the prediction of human PK from preclinical studies, but the observed AUC was approximately three‐fold higher than the preclinical prediction and thus explains the discrepancy between the predicted and observed target coverage. Based on PK/PD modeling, the estimated plasma 90% inhibitory concentration (IC 90 ) value of ~1100 ng/mL (~2.5 μM) for IL‐1β and IL‐18 inhibition was in good agreement with the in vitro human whole blood IC 90 value (639 ng/mL, 1.48 μM) after correcting for the blood:plasma ratio of 0.6 9 . Based on the observed PDs and the estimated clinical IC 90 , twice‐daily dosing of 150 mg GDC‐2394 or greater is expected to provide at least 90% target inhibition in peripheral blood throughout the dosing interval.…”

“…GDC‐2394 is an orally available, selective, and reversible small‐molecule inhibitor of NLRP3 9 . It selectively blocks IL‐1β and IL‐18 secretion following activation of the NLRP3 pathway by various stimuli, including cholesterol crystals, in biochemical and ex vivo cellular assays.…”

“…It selectively blocks IL‐1β and IL‐18 secretion following activation of the NLRP3 pathway by various stimuli, including cholesterol crystals, in biochemical and ex vivo cellular assays. Clinical development of GDC‐2394 was initiated based on nonclinical safety and pharmacology data targeting CAD as the lead indication 9 . This first‐in‐human, dose‐escalation study evaluated the safety, tolerability, pharmacokinetics (PKs), and pharmacodynamics (PDs) of single and multiple oral doses of GDC‐2394, and evaluated the effect of food and the CYP3A4 induction potential of GDC‐2394 in healthy volunteers.…”

JT002, a small molecule inhibitor of the NLRP3 inflammasome for the treatment of autoinflammatory disorders

“…Importantly, JT002 displayed good selectivity towards inhibiting NLRP3 in ammasome activation and did not interfere with either TLR4-mediated activation of the NF-κB pathway or other in ammasomes tested, such as NLRP1, NLRC4 or AIM2. Although the direct target of JT002 has not yet been identi ed, a recent publication that utilized cryo-EM demonstrated direct binding of a structurally similar compound to an allosteric pocket in the NACHT domain of NLRP3, thus explaining the in ammasome selectivity pro le 55 .…”

Novel Aryl Sulfonamide Derivatives as NLRP3 Inflammasome Inhibitors for the Potential Treatment of Cancer

“…These findings prompted the development of potent and selective NLRP3 inhibitors that are extensively used as pharmacological tools to elucidate possible clinical applications of NLRP3 targeting strategies. , In this rapidly expanding research field, numerous compounds are already under preclinical investigation, and few of them have reached phase I/II clinical trials. − Among these, MCC950 ( 1 , Figure ) is the most studied NLRP3 inhibitor after its early discovery in 2003 . MCC950 was shown to block canonical and noncanonical NLRP3 activation at nanomolar concentrations in vitro with high selectivity over AIM2, NLRC4, or NLRP1 inflammasomes .…”