Wang Wenchu, Qin Tingsheng, Feng Xianjing, Xu Mengyang, Ling Zhen, Xie Xin, Zhang Min, Huang Yanqin, Luo Jiayuan, Cao Sisi, Yue Xiaoyang, Zhang Jie, Deng Fengyuan, He Linhong
Guangxi Key Laboratory of Bioactive Molecules Research and Evaluation, Key Laboratory of Micro-Nanoscale Bioanalysis and Drug Screening, Education Department of Guangxi Zhuang Autonomous Region, Pharmaceutical College, Guangxi Medical University, Nanning, Guangxi, China.
Key Laboratory of Basic Research on Regional Diseases, College of Basic Medical Science, Guangxi Medical University, Guangxi, China.
Arch Pharm (Weinheim). 2025 Jul;358(7):e70038. doi: 10.1002/ardp.70038.
Inducible nitric oxide synthase (iNOS) is an attractive target to develop drugs for the treatment of rheumatoid arthritis (RA), whose overexpression leads to the release of abundant NO. In this study, a series of compounds were designed and synthesized bearing the scaffold of 7H-pyrrolo[2,3-d]pyrimidine to improve CLogP values and in vivo anti-inflammatory activity via molecular hybridization, whose derivatives in a previous study demonstrated good inhibition toward NO production but high CLogP values and weak potency in vivo. Among them, N3b effectively suppressed NO production in a concentration-dependent manner (IC = 4.66 ± 0.19 μM) in LPS-induced RAW264.7 cells. Mechanically, N3b mildly inhibited the protein expression of iNOS (IC = 13.60 ± 0.39 μM), without remarkable influence on the mRNA levels of iNOS or other classical pro-inflammatory cytokines including IL-1β, IL-6, and TNF-α at the high concentration of 30 µM. With favorable drug-likeness, N3b significantly alleviated paw edema at the dose of 30 mg/kg in the acute inflammation model and mitigated paw swelling and joint damage at a dosage of 10 mg/kg in the chronic arthritis model. Furthermore, N3b did not cause any obvious weight loss and pathological changes of important organs in treated mice, indicating that N3b is a potent candidate iNOS inhibitor for the treatment of RA.
诱导型一氧化氮合酶（iNOS）是开发治疗类风湿性关节炎（RA）药物的一个有吸引力的靶点，其过度表达会导致大量一氧化氮的释放。在本研究中，设计并合成了一系列以7H-吡咯并[2,3-d]嘧啶为骨架的化合物，通过分子杂交提高其脂水分配系数（CLogP）值和体内抗炎活性，其在先前研究中的衍生物对一氧化氮的产生具有良好的抑制作用，但CLogP值较高且体内效力较弱。其中，N3b在脂多糖（LPS）诱导的RAW264.7细胞中以浓度依赖性方式有效抑制一氧化氮的产生（IC = 4.66 ± 0.19 μM）。机制上，N3b轻度抑制iNOS的蛋白表达（IC = 13.60 ± 0.39 μM），在30 μM的高浓度下对iNOS或其他经典促炎细胞因子（包括IL-1β、IL-6和TNF-α）的mRNA水平没有显著影响。N3b具有良好的类药性，在急性炎症模型中以30 mg/kg的剂量显著减轻爪部水肿，在慢性关节炎模型中以10 mg/kg的剂量减轻爪部肿胀和关节损伤。此外，N3b在治疗的小鼠中未引起任何明显的体重减轻和重要器官的病理变化，表明N3b是治疗RA的一种有效的iNOS抑制剂候选物。
