Natural polycyclic aromatic naphthodianthrone compound S3273 calms cytokine storm to restore acute lung injury via modulating CD39-P2X7R-IL-17A route.

CD39 participates in the process of ATP hydrolyzing into AMP, ADP and adenosine, and ATP mediates the channel-opening of P2X7 which further induces the downstream pyroptotic cell death. Current announced small-molecule CD39 enhancer are limited and to develop novel CD39 enhancer will be meaningful for inflammation-associated disorders treatment. CD39 enzymic screening assay was conducted to detect CD39 enhancing effect among a library of 424 FDA-approval small-molecule compounds. CETSA and SPR assay were arranged to assess the direct interaction between S3273 and CD39. Multiple techniques like YO-PRO-1 assay and patch clamp were employed to evaluate diverse CD39-P2X7 route modulators. RNA-Seq experiment was thereafter employed to fish potential targets in LPS-challenged mice. A polycyclic aromatic naphthodianthrone compound, S3273, was the most potent CD39 enhancer with EC50 value of 5.66 μM. S3273 restored LPS-induced ALI via blocking NLRP3/GSDMD signaling to calm cytokine storm. In vitro, S3273 rescued pyroptosis via inhibiting the ROS accumulation, inflammasome assembly, as well as pyroptosis in a range types of macrophages. Mechanically, S3273 affected purinergic modulators including CD39, CD73, A2b and P2X7R to facilitate the formation and function of CD39-P2X7-dominated purinergic brake. IL-17 was a key pathway responsible for the protective effect of S3273 on LPS-induced ALI. S3273 directly binds to IL-17 and prevented the up-regulation of IL-17. S3273 calms cytokine storm to reverse lung damage in LPS-challenged mice, and these effects are at least partly through the modulation on CD39-P2X7R-IL-17A route.