圆弧菌醇和圆弧菌素的分离纯化及结构解析

肿瘤坏死因子α(TNF-α)是激活的巨噬细胞分泌的一种多功能因子,其过量表达可以引起类风湿性关节炎和其他自身免疫性疾病.因此,寻找开发其小分子抑制剂具有重要意义.在对一株分离自厦门海沧红树林土壤青霉属(Peni-cilliumsp.)真菌的次级代谢产物进行分离纯化以获得TNF-α抑制剂的研究中,通过中压反相硅胶柱层析(RP-18)、常压凝胶柱层析(Sephadex LH-20)、硅胶柱层析(Si-Gel)等柱层析技术对两种产量较高的化合物进行了分离纯化.通过对两个化合物的一维(1D-)、二维(2D-)核磁共振(NMR)和质谱(MS)数据的深入解析,确定两个化合物为生物碱圆弧菌醇和圆弧菌素,两者的产率分别达7.0和3.9mg/L,并通过X射线单晶衍射确定了圆弧菌醇手性碳的绝对构型.圆弧菌醇和圆弧菌素是生物合成TNF-α诱导的人类HIV病毒复制强抑制剂纯绿青霉素过程的中间产物,但是在拮抗过量TNF-α的实验中并没有表现出明显的活性.该研究为开发利用这两种生物碱及通过生物转化合成纯绿青霉素,及加速新的潜在免疫抑制剂的研究提供基础.

Purifications and Structure Elucidations of Cyclopenol and Cyclopenin

Over-expression of the tumor necrosis factor alpha(TNF-α) is responsible for a number of pathological conditions,and TNF-α plays a pivotal role in the origin and progression of rheumatoid arthritis(RA) and other immune mediated disorders,so the development of small-molecule TNF-α inhibitors is significant.In the course of screening the inhibitors of TNF-α from secondary metabolites of microorganisms isolated from marine,two benzodiazepine alkaloids were obtained from the fungus Penicllium sp.,which was isolated from a soil sample collected from Haicang mangrove,Xiamen,China.They were purified with column chromatographies,especially silica gel RP-18,sephadex LH-20 and silica-gel column chromatography.On the basis of thorough analysis of NMR and MS spectra,they were identified as cyclopenol and cyclopenin with high-yield of 7.0 and 3.9 mg/L,respectively.Cyclopenol was further confirmed by a single crystal X-ray diffraction analysis.Cyclopenin and cyclopenol can be converted into the quinolines viridicatin,wich is strong inhibitors of the TNF-α-induced replication of human HIV.Although they showed no significant inhibitory activity of the excessive TNF-α,the determination of absolute configuration is significant to pharmacological study in immune and structural optimization,this maybe help to accelerate the development of potential immune inhibitors.The study of viridicatin maybe help to accelerate the development of potential immune inhibitors.