Bingqian Han,1,2,* Shiyu Ma,3,* Wenjin Liu,2,* Yuyang Wang,2,* Mingzhu Wang,1 Yuanzhe Li,1 Chuanjun Song,3 Yongfang Yao,1– 3 Moran Sun,2 Yongtao Duan1 

1Henan Provincial Key Laboratory of Pediatric Hematology, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou University, Zhengzhou, Henan, People’s Republic of China; 2School of Pharmaceutical Science, Zhengzhou University, Zhengzhou, Henan, People’s Republic of China; 3College of Chemistry, Pingyuan Laboratory, Zhengzhou University, Zhengzhou, Henan, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Yongtao Duan, Henan Provincial Key Laboratory of Pediatric Hematology, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou University, Zhengzhou, Henan, 450018, People’s Republic of China, Email duanyongtao860409@163.com Moran Sun, Henan Provincial Key Laboratory of Pediatric Hematology, Children’s Hospital Affiliated to Zhengzhou University, Zhengzhou University, Zhengzhou, Henan, 450018, People’s Republic of China, Email sunmr@zzu.edu.cn

Purpose: The P2Y14 receptor (P2Y14R) is closely associated with several inflammatory diseases in humans. Although several P2Y14R antagonists have been reported to date, few have been successfully developed as therapeutic drugs, and none have entered clinical trials. We aimed to obtain P2Y14R antagonists with high antagonistic activity and druggability for further investigation into anti-inflammatory drugs.
Methods: Three series of novel P2Y14R antagonists were screened. The druggability of the most promising compounds was evaluated through assays for the inhibition of cytochrome P450 and hERG (human Ether-à-go-go-Related Gene) channels, as well as pharmacokinetic experiments. The in vivo efficacy of the lead compound was assessed in a Lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model.
Results: We designed a series of N-acyl tryptophan derivatives as novel and potent P2Y14R antagonists based on the hit compound 7. Among them, compound II-3 with an IC50 value of 1.2 nM, was a better antagonist than PPTN with an IC50 value of 2.0 nM. Through structural modification, the zwitterionic character was eliminated, resulting in significantly improved solubility and oral bioavailability compared to PPTN. We have confirmed that P2Y14R is highly expressed in macrophages of ALI lung tissue. II-3, as a P2Y14R antagonist, can alleviate the pathological progression of ALI by inhibiting the activation of the NLRP3 inflammasome pathway and reducing the release of inflammatory factors, thus providing direct evidence for P2Y14R as a therapeutic target.
Conclusion: Compound II-3 with potent P2Y14R antagonistic activity, may be a promising candidate for further investigation as an anti-inflammatory drug.

Keywords: P2Y14 receptor, antagonist, G protein-coupled receptor, N-acyl tryptophan derivatives, anti-inflammatory activity