以DNA回旋酶B亚基为靶点的抗结核药物筛选模型的建立及初步应用

Study and primary application of the screening model for antituberculosis drugs with DNA gyrase B subunit as a target

实验采用Mycobacteriumphlei为出发菌株,利用NTG和紫外线复合诱变得到新生霉素特异性的超敏菌株M.phleiSS12和耐药菌株M.phleiRR11,建立了以DNA回旋酶B亚基为靶点的抗结核药物的筛选模型;并应用此模型对1600株真菌和100株放线菌发酵液进行了初筛,获得了38株阳性菌,阳性率为2.4%;对阳性菌株3277的发酵液进行了提取分离,得到X1和X2两个活性化合物。根据两个化合物的分子量和特征性紫外吸收,经MPMS数据库及PubMed联机检索,认为X1、X2可能为新化合物。

In order to find new kinds of antibacterial agents, a primary screening model has been set with the DNA gyrase B subunit as a target, and then the active compounds of positive strain were purified. From Mycobacterium phlei, we isolated a novobiocin supersensitive mutant M.phlei SS12 and a resistant ~mutant M.phlei RR11 with NTG and UV. The screening method was proved by testing the inhibitions to those three strains of a series of drugs which have different action mechanisms with novobiocin. During the primary screening process, M.phlei, M.phlei SS12 and M.phlei RR11 were used as test strains. From 1600 species of fungi and 100 species of actinomyces fermentation broth, we got 38 positive strains. In these strains, 3277 which had fairly stabilized fermentation character and show potent inhibitory activity against M.phlei was picked up and tested on further study. Two active compounds, named X1 and X2, were purified from the ~fermentation broth of strain 3277 by solvent extraction, silica gel column chromatography, sephadex LH-20 column chromatography and ODS chromatography. According to their characteristic UV λ_~max (MeOH) and molecular weights, it was designated that they are new chemical entities.