4-酰胺基苯并噁唑酮类抗炎小分子化合物的合成及其对MD2蛋白的抑制活性研究  

Synthesis and in vitro anti-inflammatory activity assay of 4-acylamino-benzoxazolone derivatives as myeloid differentiation protein 2(MD2)inhibitor

作　　者：白慧莹 高晓慧 赵蓓 唐莉[1] 李青山[1,2] BAI Hui-ying;GAO Xiao-hui;ZHAO Bei;TANG Li;LI Qing-shan(School of Pharmaceutical Science,Shanxi Medical University,Taiyuan 030001,China;Shanxi Key Laboratory of Innovative Drug for the Treatment of Serious Diseases Basing on the Chronic Inflammation,Shanxi University of Traditional Chinese Medicine,Taiyuan 030024,China)

机构地区：[1]山西医科大学药学院,山西太原030001 [2]山西中医药大学基于炎性反应的重大疾病创新药物山西省重点实验室,山西太原030024

出　　处：《中国药物化学杂志》2022年第3期178-184,共7页Chinese Journal of Medicinal Chemistry

基　　金：国家自然科学基金项目(81602976);山西省青年科技研究基金项目(201901D211350);山西省回国留学人员重点科研项目(2014-重点2)。

摘　　要：目的设计合成系列4-酰胺基苯并噁唑酮类化合物,并测定其体外抗炎活性,探究其与MD2蛋白的结合能力,以期发现高抗炎活性的新型MD2抑制剂。方法以3-硝基-2-氨基苯酚为原料,经过成环、还原、取代反应制得目标化合物,通过ESI-MS、^(1)H-NMR、^(13)C-NMR对其结构进行确证。建立脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7炎症模型,Griess法测定细胞培养液中NO的释放量,ELISA法测定炎症因子IL-6的含量,评价化合物的体外抗炎活性;流式细胞术、bis-ANS小分子荧光光谱技术结合计算机模拟检测高活性化合物与MD2蛋白的结合能力及其作用方式。结果与结论合成了9个未见文献报道的新化合物,其结构均经ESI-MS、^(1)H-NMR和^(13)C-NMR确证。大多数化合物对NO、IL-6呈现出较好的抑制活性。尤其是化合物4b,在浓度为25μmol·L^(-1)时对NO抑制率为(72.5±3.16)%,对IL-6的抑制率为(48.5±2.83)%,与阳性药塞来昔布[NO抑制率(88.9±2.22)%,IL-6抑制率(45.1±3.90)%]相当;化合物4b可剂量依赖性竞争LPS与细胞膜表面受体的结合,竞争bis-ANS小分子荧光探针与rhMD-2蛋白的结合,是新型具有抗炎活性的MD2抑制剂。Compound 4-(5'-dimethylamino-naphthalene)-sulfonyloxy-benzoxazolone(W3D)was a new benzoxazolone derivative with high anti-inflammatory activity obtained in our research group.Previous research has presented that W3D could directly bind to MD2 protein which might be a new MD2 inhibitor.In order to improve the structure-activity relationship and find the integration mode with MD2 protein,nine 4-acylamino-benzaxazolone derivatives were designed and synthesized via cyclization,reduction and substitution reaction using 3-nitro-2-aminophenol as the starting material.All the target compounds were characterized by ESI-MS,^(1)H-NMR and ^(13)C-NMR spectra.The anti-inflammatory activity in LPS induced RAW264.7 cell showed that most of the compounds presented certain inhibitory activities against NO and IL-6,and compound 4b exhibited the highest activity which was comparable to the positive drug celecoxib.Meanwhile,compound 4b could competitively prevent the binding of LPS to membrane surface receptors and inhibit the binding of fluorescent probe bis-ANS to rhMD2 protein in a dose dependent manner.

关 键 词：苯并噁唑酮 合成 抗炎 MD2抑制剂 

分 类 号：R914[医药卫生—药物化学]