To develop next-generation Pannexin 1 modulators as new strategies for functional tissue repair and remodeling

Pannexin 1 (PANX1) is a large-pore ion channel that plays a crucial role in various physiological processes such as apoptosis and purinergic signaling. However, commonly used PANX1 inhibitors generally suffer from low selectivity and poor efficacy, leading to potential misinterpretations in basic research and limiting their applications in both fundamental and translational studies. Therefore, there is an urgent need to develop novel inhibitors with high specificity and potency. Building upon our previous work, this study successfully developed a series of novel naphthyridone derivatives with excellent PANX1 inhibitory activity and, for the first time, elucidated their molecular mechanisms. Through drug synthesis and optimization, combined with flow cytometry-based screening and electrophysiological assays, we identified small-molecule inhibitors with significantly lower IC₅₀ values compared to traditional agents, without affecting LRRC8/SWELL1 channels or eukaryotic topoisomerase II, thus addressing previous concerns regarding cell volume regulation and hepatotoxicity. Furthermore, by integrating structure-activity relationship analysis, site-directed mutagenesis, and molecular docking simulations, we defined the key functional groups and critical binding sites on PANX1, demonstrating that the new inhibitors maintain robust inhibition under varying membrane potentials, thereby laying the groundwork for further investigation into PANX1 gating mechanisms. Finally, using a DSS-induced murine colitis model, we provided the first evidence that PANX1 inhibition can effectively alleviate intestinal inflammation and disease severity, establishing PANX1 as a promising therapeutic target not only for inflammatory bowel diseases but also for other immune-related conditions such as chronic neuropathic pain, brain injury, and ischemia-reperfusion damage.