HDL-16 is a potent P2Y14R antagonist with an IC50 of 0.3095 nM, which ameliorates DSS-induced colitis by suppressing necroptosis of intestinal epithelial cells (IECs) and protecting mucosal barrier function [1].

P2Y14 Receptor:0.3095 nM

HDL-16 forms a hydrogen bond between the N atom on its benzooxazole ring and the Tyr102 residue, while its amino group also interacts through hydrogen bonding with the His184 residue in the binding pocket of P2Y14R [1]. At a concentration of 10 μM, HDL-16 exhibits negligible cytotoxicity towards HT-29 cells [1]. Additionally, HDL-16 can inhibit necroptosis in HT-29 cells (induced by 20 ng/mL TNF-α, 20 μM z-VAD-fmk, and the Smac mimetic BV6 at 2 μM, for 8 hours) [1].

Administering a high dosage of HDL-16 (10 µM or 20 µM; 100 µL; rectal administration; daily; for 6 days) significantly mitigated symptoms of colitis in mice and preserved the integrity of the mouse intestinal barrier [1]. The experimental animal models were male P2Y14R fl/fl Vil-cre (P2Y14R ∆IEC), P2Y14R fl/fl Lyz2-cre, and WT mice with a C57BL/6J background, aged 7-8 weeks [1]. Administration of 10 µM or 20 µM HDL-16 doses daily via rectal delivery over six days showed that the high-dose group experienced significantly less weight loss and lower disease activity index (DAI) compared to mice exposed to 3% w/v, 36-50 kDa DSS (via drinking water for 7 days). The higher doses improved DSS-induced inflammatory infiltration and tissue damage and significantly reduced epithelial cell necrosis in the intestines of DSS-treated mice.