海洋链霉菌7-145中洋橄榄叶素衍生物的分离鉴定  

Isolation and characterization of elaiophylins from the marine-derived Streptomyces sp. 7-145

作　　者：吴春彦[1,2] 谭亿[2] 甘茂罗[2,3] 蒙建州[2] 关艳[2] 周红霞[2] 戴剑漉[2] 王以光[2] 尚小雅[1] 杨兆勇[2] 肖春玲[2] 

机构地区：[1]北京联合大学生物活性物质与功能食品北京市重点实验室,北京10083 [2]中国医学科学院北京协和医学院医药生物技术研究所,北京100050 [3]天然药物活性物质与功能国家重点实验室,中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《中国抗生素杂志》2014年第3期186-192,共7页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(81273414;81001386);天然药物活性物质与功能国家重点实验室开放课题(GTZK201302)

摘　　要：目的分离鉴定海洋链霉菌Streptomyces sp．7-145产生的抗耐药菌活性成分。方法通过活性追踪，利用大孔吸附树脂、凝胶、正反相色谱等多种色谱技术分离纯化活性成分；通过紫外光谱、质谱、核磁共振谱等波谱学方法鉴定其结构。利用琼脂稀释法和微量肉汤稀释法分别测定所得化合物的抗耐药菌和抗结核活性，利用MTT法评价其细胞毒性。结果从中分离鉴定了4个活性化合物，其结构分别确定为洋橄榄叶素（1）、11-O-甲基洋橄榄叶素（2）、11，11’-O-二甲基洋橄榄叶素（3）、14’-去乙基-14＇-甲基洋橄榄叶素（4）。化合物1～4对甲氧西林耐药的金黄色葡萄球菌（MRSA）和万古霉素耐药的肠球菌（VRE）等革兰阳性耐药菌具有较强的抗菌活性，最低抑菌浓度（MIC）为1～16μg／mL；所有化合物对结核分枝杆菌H37Rv株显示出显著的抑制活性（MIC：0．5～4μg／mL），其中化合物1对结核分枝杆菌多重耐药（MDR）、广泛耐药（XDR）等耐药株也表现出较强的抑制活性（MIC：0．5～1μg／mL）；1和2对人肝细胞（L02）、肾上皮细胞（293T）和乳腺癌细胞（MCF-7）具有较强的细胞毒活性，IC50值200．14～0．28μmol／L。结论化合物1～4具有较强的体外抗MRSA、VRE等耐药菌活性。首次发现洋橄榄叶素（1）具有体外抗结核分枝杆菌标准株、多重耐药以及广泛耐药株活性。化合物1和2对人正常肝、肾上皮细胞具有细胞毒性。Objective To isolate and identify the antibacterial metabolites against drug-resistant pathogens from the fermentation broth of Streptomyces sp. 7-145. Methods A PCR-based genetic screening experiment targeting the dTDP-glucose-4,6-dehydratase gene revealed that Streptomyces sp. 7-145 had the potential to produce glycosidic antibiotics. Chemical investigation of culture extracts of the strain was performed using a combination of chromatographic techniques including macroporous resin, silica gel, reverse-phase HPLC under the guidance of bioassay. Structures of the isolated compounds were determined by using spectroscopic methods including UV, 1D-NMR, 2D-NMR and MS spectra. Results Four compounds were isolated from the fermentation broth of Streptomyces sp. 7-145. Their structures were elucidated as elaiophylin （1）, 11-O-methylelaiophylin （2）, 11,11＇-O-dimethylelaiophylin （3）, and efomycin G （4）. Compounds 1-4 showed good antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus au reus （MRS A） and vancomycinresistant enterococci （VRE） strains with the minimal inhibitory concentration （MIC） values ranging from 1 to l oug/ml., All compounds displayed potent activities against Mycobacterium tuberculosis H37Rv with MIC values of 0.5-4μg/mL. Compound 1 also showed good activity againt clinically isolated multi drug-resistant （MDR） and extensively drug-resistant （XDR） M. tuberculosis strains （MIC： O.5-1μg/mL）. In addition, compounds 1 and 2 showed significant cytotoxicities against human liver cell （L02）, kidney epithelial cell （293T） and mammary adenocarcinoma cell （MCF- 7） with ICso values in the range of 0.14-0.28μg/mL. Conclusion Four elaiophylins with potent activities against MRSA and VRE strains were identified from Streptomyces sp. 7 -145. The antibacterial activities of compounds 1 -4 againt M. tuberculosis were reported for the first time.

关 键 词：海洋放线菌 链霉菌 洋橄榄叶素 结核分枝杆菌 耐药菌 

分 类 号：R379.1[医药卫生—病原生物学]