人高密度脂蛋白受体CLA-1表达上调剂4776B和4776C的分离、结构鉴定及生物活性研究  

Study of isolation,identification and biological activity of human high density lipoprotein receptor CLA-1 up-regulator 4776B and 4776C from Actinomycete 04-4776

作　　者：杨媛[1] 张忠兵[2] 姜威[2] 樊秀勇[2] 王旻[1] 张月琴[2] 洪斌[2] 司书毅[2] 

机构地区：[1]中国药科大学生命科学与技术学院,南京210009 [2]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《中国抗生素杂志》2007年第5期268-272,276,共6页Chinese Journal of Antibiotics

基　　金：国家自然科学基金资助项目(No.30472026)

摘　　要：目的从微生物代谢产物中筛选和分离能够上调人高密度脂蛋白受体(CLA-1)表达的新型活性化合物。方法应用本实验室已经建立的CLA-1表达上调剂的高通量筛选模型,从6000多株微生物中筛选出8株能使CLA-1表达上调的阳性菌株。对其中一株阳性菌株放线菌04-4776发酵产物的活性成分进行活性跟踪分离纯化,从中获得CLA-1表达上调的化合物4776B和4776C,并测定了化合物的理化特性和波谱学数据及相关的生物活性。结果通过理化性质、质谱、紫外和核磁等波谱学数据的分析,确定了化合物4776B和4776C的结构;4776B和4776C对CLA-1的上调率分别在2.08和3.26μm ol/L时达到最大,与对照相比分别上调了60%和78%。结论放线菌04-4776产生的两个活性化合物分别与文献报道的红车轴草素和(9R,13S)-7-脱氧-13-二氢道诺霉素酮结构一致;两个化合物均具有强的上调CLA-1的表达活性,属首次报道。Objective To screen and isolate new compounds which can up-regulate human high density lipoprotein receptor CLA-1 expression from microbial secondary metabolites. Method Using the high throughput screening model in vitro, 8 strains from six thousand actinomycetes are shown to produce up-regu- lators of CLA-1. Actinomycete strain 04-4776 was chosen for further isolation and characterization of active compounds. The fermentation broth of actinomycete strain 04-4776 was used to isolate active compounds which might increase CI.A-1 expression. By column chromatography of HP20 macroporous resin, I.H20 sephadex gel filter and semi-preparative HPLC, two pure compounds 4776B and 4776C were isolated from the broth. The properties and spectral data of the two compounds were determined and the biological activities were analyzed. Results By means of EI-MS, ESI-MS, ^1H, ^13C and 2D NMR spectra, the structure of 4776B was identified as 4＇ - methoxyl - 3 ＇ , 5 , 7 - hydroxyl isoflavone and the structure of 4776C was identified as （9R, 13S）-7 - deoxy - 13-dihydro-daunomycinone. The maximal CLA-1 up-regulatory activity of 4776B reached to 60% at about 2. 08μmol/L while that of 4776C reached to 78% at about 3. 26μmol/L. Conclusion Compounds 4776B and 4776C produced by Actinomycete 04-4776 were identical as pratensein and （9R, 13S）-7-deoxy-13-dihydro-dauno- mycinone, respectively. The CLA-l-up-regulating bioactivity of these two compounds is first reported.

关 键 词：人高密度脂蛋白受体 上调剂 高通量筛选 微生物次级代谢产物 

分 类 号：Q939.9[生物学—微生物学]