Hwang Tsong-Long, Wang Wen-Hui, Wang Ting-Yi, Yu Huang-Ping, Hsieh Pei-Wen
Graduate Institute of Natural Products, School of Traditional Chinese Medicine, and Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan; Chinese Herbal Medicine Research Team, Healthy Aging Research Center, Chang Gung University, Taoyuan, Taiwan; Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kweishan, Taoyuan, Taiwan.
Graduate Institute of Natural Products, School of Traditional Chinese Medicine, and Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
Bioorg Med Chem. 2015 Mar 1;23(5):1123-34. doi: 10.1016/j.bmc.2014.12.056. Epub 2015 Jan 16.
Proteinase 3 (Pr3), and human neutrophil elastase (HNE) are two major neutrophilic serine proteases (NsPs) expressed in neutrophil azurophil granules. Emerging data suggest that excessive release of proteases mediates tissue damage, and therefore prolonged neutrophil accumulation has an important role in the pathogenesis of many diseases. Thus, HNE and Pr3 inhibitors may prove to be targets for the generation of agents in the treatment of neutrophilic inflammatory disease. Sivelestat is the only commercially available selective HNE inhibitor. Therefore, sivelestat was chosen as the model structure in an attempt to obtain more potent anti-NsPs agents. In the present study, a series 2-aminobenzaldehyde oxime and 2-aminobenzoate analogs were synthesized and their inhibitory effects on NsPs (CatG, Pr3, and HNE) were determined, respectively. The results of structure-activity relationships studies concluded that a hydroxyl oxime moiety plays an important role in ligand-enzyme affinity through hydrogen bonding. As compound 6 had more potency and showed dual inhibitory effects on NE and Pr3, both in vitro and in vivo experiments were carried out to evaluate its selectivity, effects in cell-based assays, and efficacy in models of inflammation and damage. Compound 6 had the potential to reduce paw edema induced by LPS and HNE, as well as acute lung injury, and may be approved as a candidate for the development of new agents in the treatment of neutrophilic inflammatory diseases.
蛋白酶3（Pr3）和人中性粒细胞弹性蛋白酶（HNE）是在中性粒细胞嗜天青颗粒中表达的两种主要的嗜中性粒细胞丝氨酸蛋白酶（NsPs）。新出现的数据表明，蛋白酶的过度释放介导组织损伤，因此中性粒细胞的长期积聚在许多疾病的发病机制中起重要作用。因此，HNE和Pr3抑制剂可能被证明是治疗嗜中性粒细胞炎性疾病药物研发的靶点。西维来司他是唯一可商购的选择性HNE抑制剂。因此，选择西维来司他作为模型结构，试图获得更有效的抗NsPs药物。在本研究中，合成了一系列2-氨基苯甲醛肟和2-氨基苯甲酸酯类似物，并分别测定了它们对NsPs（组织蛋白酶G、Pr3和HNE）的抑制作用。构效关系研究结果表明，羟肟基团通过氢键在配体-酶亲和力中起重要作用。由于化合物6具有更高的效力，并且在体外和体内实验中均对NE和Pr3表现出双重抑制作用，因此进行了体外和体内实验以评估其选择性、在基于细胞的实验中的作用以及在炎症和损伤模型中的疗效。化合物6有潜力减轻由脂多糖和HNE诱导的爪肿胀以及急性肺损伤，并且可能被批准作为治疗嗜中性粒细胞炎性疾病新药研发的候选药物。
