桦木酸磺酰胺衍生物类小分子Omicron融合抑制剂的综合性实验设计  

Comprehensive experimental design for betulinic acid sulfonamide derivatives as novel SARS-CoV-2 Omicron small-molecule fusion inhibitor

作　　者：宋高鹏 刘铭健 王进绅 倪春林 刘叔文 SONG Gaopeng;LIU Mingjian;WANG Jinshen;NI Chunlin;LIU Shuwen(Biochemical Engineering and Pharmaceutical Experimental Teaching Demonstration Center,College of Materials and Energy,South China Agricultural University,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of New Drug Screening,School of Pharmaceutical Sciences,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]华南农业大学材料与能源学院生物化工与制药广东省实验教学示范中心,广东广州510642 [2]南方医科大学药学院广东省新药筛选重点实验室,广东广州510515

出　　处：《实验技术与管理》2023年第9期102-108,124,共8页Experimental Technology and Management

基　　金：国家自然科学基金项目(82073722);广东省基础与应用基础研究基金项目(2022A1515010016);广东省本科高校教学质量与教学改革工程建设项目“生物化工与制药实验教学示范中心(2017)”和“制药工程专业系列实践课程教学团队(2019)”。

摘　　要：新冠病毒刺突蛋白的S2介导了病毒与宿主细胞的膜融合过程,其氨基酸酸序列在Omicron等变异株中均高度保守,可作为开发新型抗新冠病毒药物的重要靶点。基于此前沿热点课题,该文设计了一类以桦木酸磺酰胺皂苷衍生物为结构骨架的新型小分子Omicron融合抑制剂的综合性实验。以马铃薯三糖桦木酸皂苷1为苗头化合物,利用生物电子等排体原理及基于结构的药物设计方法(SBDD)设计并合成了三个马铃薯三糖桦木酸磺酰胺衍生物T-1、T-2、T-3,利用NMR及HRESIMs对其进行结构表征,并基于S/HIV模型测试了其在细胞水平对Omicron等多种新冠病毒变异株的抑制活性。在此基础上,综合利用表面等离子共振SPR、免疫共沉淀(Co-PI)、细胞-细胞融合实验、分子对接等实验得出该类皂苷分子能够嵌入S1/S2亚基交界处空腔、稳定S蛋白融合前的构象从而抑制病毒入胞,进而表现出广谱抗新冠病毒活性的结论。The S2 subunit of Omicron spike protein is responsible for regulating S-mediated viral/cell membrane fusion to catalyze the virus entry into hose cell step,of which amino acid sequences are highly conserved.Thus,the S2 protein is an important target for development of anti-SARS-CoV-2 drugs.Based on this frontier topics,we designed a comprehensive experiment including the preparation and evaluation of betulinic acid sulfonamide saponins derivatives against SARS-CoV-2 Omicron entry into host cells.In the experiment,the 3-O-β-chacotriosyl betulinic acid saponin 1 was chosen as the hit compound,of which sulfonamide derivatives T1,T-2,T-3 were designed on the basis of the bioisosterism strategy and the SBDD method.The structures of all the title compounds were confirmed by 1HNMR,13CNMR and HRESIMs.After screening of anti-SARS-CoV-2 activity in vitro using pOmicron S/HIV,the preliminary anti-virus mechanism demonstrated that these betulinic acid saponins derivatives could copy the pocket located at the junction of S1 and S2 subunits in the spike protein surface based on the SPR,Co-PI,the spike-dependent cell-cell fusion assay and docking studies,which was helpful in stabling the prefusion conformation of S to block SARS-CoV-2 viruses into host cells,thereby showing broad-spectrum anti-SARS-CoV-2 activities in vitro.

关 键 词：实验教学 新冠病毒小分子融合抑制剂 桦木酸磺酰胺衍生物 综合实验 

分 类 号：R914[医药卫生—药物化学] G642[医药卫生—药学]