  Inspired by the recent advancements in understanding the binding mode of
sulfonylurea-based NLRP3 inhibitors to the NLRP3 sensor protein, we developed
new NLRP3 inhibitors by replacing the central sulfonylurea moiety with
different heterocycles. Computational studies evidenced that some of the
designed compounds were able to maintain important interaction within the NACHT
domain of the target protein similarly to the most active sulfonylurea-based
NLRP3 inhibitors. Among the studied compounds, the 1,3,4-oxadiazol-2-one
derivative 5 (INF200) showed the most promising results being able to prevent
NLRP3-dependent pyroptosis triggered by LPS/ATP and LPS/MSU by 66.3 +/- 6.6%
and 61.6 +/- 11.5% and to reduce IL-1\b{eta} release (35.5 +/- 8.8 % {\mu}M) at
10 {\mu}M in human macrophages. The selected compound INF200 (20 mg/kg/day) was
then tested in an in vivo rat model of high-fat diet (HFD)-induced
metaflammation to evaluate its beneficial cardiometabolic effects. INF200
significantly counteracted HFD-dependent "anthropometric" changes, improved
glucose and lipid profiles, and attenuated systemic inflammation and biomarkers
of cardiac dysfunction (particularly BNP). Hemodynamic evaluation on
Langendorff model indicate that INF200 limited myocardial damage-dependent
ischemia/reperfusion injury (IRI) by improving post-ischemic systolic recovery
and attenuating cardiac contracture, infarct size, and LDH release, thus
reversing the exacerbation of obesity-associated damage. Mechanistically, in
post-ischemic hearts, IFN200 reduced IRI-dependent NLRP3 activation,
inflammation, and oxidative stress. These results highlight the potential of
the novel NLRP3 inhibitor, INF200, and its ability to reverse the unfavorable
cardio-metabolic dysfunction associated with obesity.
