Anti-inflammatory Effects of Novel P2X4 Receptor Antagonists, NC-2600 and NP-1815-PX, in a Murine Model of Colitis

• Published in: Inflammation 2022-08, Vol.45 (4), p.1829-1847
• Main Authors: D'Antongiovanni, Vanessa, Pellegrini, Carolina, Benvenuti, Laura, Fornai, Matteo, Di Salvo, Clelia, Natale, Gianfranco, Ryskalin, Larisa, Bertani, Lorenzo, Lucarini, Elena, Di Cesare Mannelli, Lorenzo, Ghelardini, Carla, Nemeth, Zoltan H, Haskó, György, Antonioli, Luca
• Format: Article
• Language: English
• Subjects: Animals; Anti-Inflammatory Agents - pharmacology; Anti-Inflammatory Agents - therapeutic use; Azepines; Caspase 1; Colitis - chemically induced; Colitis - drug therapy; Colitis - pathology; Disease Models, Animal; Inflammasomes - metabolism; Inflammation - drug therapy; Inflammation - metabolism; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Original; Oxadiazoles; Purinergic P2X Receptor Antagonists
• ISSN: 1573-2576; 0360-3997; 1573-2576
• DOI: 10.1007/s10753-022-01663-8

The pharmacological blockade of P2X4 receptors has shown potential benefits in the management of several immune/inflammatory diseases. However, data regarding the involvement of P2X4 receptors in the pathophysiological mechanisms of action in intestinal inflammation are not well defined. We aimed to evaluate the anti-inflammatory effects of two novel and selective P2X4 receptor antagonists, NC-2600 and NP-1815-PX, and characterize the molecular mechanisms of their action in a murine model of 2,4-dinitrobenzene sulfonic acid (DNBS)-induced colitis. These two drugs and dexamethasone (DEX) were administered orally for 6 days, immediately after the manifestation of DNBS. The body weight decrease, resulting from colitis, was attenuated by NC-2600 and NP-1815-PX, but not DEX. However, all three drugs attenuated the increase in spleen weight and ameliorated macroscopic and microscopic colonic tissue damage. Furthermore, all three compounds decreased tissue IL-1β levels and caspase-1 expression and activity. Colonic tissue increase of tumor necrosis factor was downregulated by DEX, while both NC-2600 and NP-1815-PX were ineffective. The reduction of occludin associated with colitis was ameliorated by NC-2600 and NP-1815-PX, but not DEX. In THP-1 cells, lipopolysaccharide and ATP upregulated IL-1β release and NLRP3, caspase-1, caspase-5, and caspase-8 activity, but not of caspase-4. These changes were prevented by NC-2600 and NP-1815-PX treatment. For the first time, the above findings show that the selective inhibition of P2X4 receptors represents a viable approach to manage bowel inflammation via the inhibition of NLRP3 inflammasome signaling pathways.