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目的研究阐明无活性放线菌野生株L35-1来源硫酸二乙酯(DES)诱变链霉素抗性抗肿瘤活性突变株D2s4-1新产代谢产物及其抗肿瘤活性。方法组合利用活性跟踪与微量预试先导-放大实验制备的实验模式,通过与原始菌样品直接对照,在快速确定差异活性斑点基础上,组合利用各种色谱技术,分离纯化突变株新产代谢产物。根据理化性质和波谱数据鉴定化合物结构。采用MTT法测定抗肿瘤活性。结果从突变株D2s4-1发酵物中分离鉴定了6个该突变株新产代谢产物,即1,9-二甲酯吩嗪(1)、2-羟基苯乙酰胺(2)、2-吡咯甲酸(3)、大豆黄素(4)、环(4-羟基-脯氨酸-亮氨酸)二肽(5)和尿嘧啶(6)。其中1、3、5和6有一定抗肿瘤活性,浓度为100μg/ml时对K562细胞的抑制率分别为33.3%、16.3%、33.3%和21.1%。结论阐明了抗肿瘤活性突变株D2s4-1的6个新产物,其中4个为活性产物。化合物1为新天然产物,其抗肿瘤活性亦属首次筛选发现。用化学诱变组合抗性筛选技术从无活性放线菌野生株转化获取的活性突变株可供筛选新产活性产物,从而拓展放线菌药源活性新菌株资源。
Objective To study the new metabolites of streptomycin-resistant and antitumor mutant D2s4-1 induced by wild-type L35-1 actinomycete and its anti-tumor activity. Methods Combining the experimental mode of activity tracking and pre-pilot-scale pilot preparation, direct comparison with the original bacteria samples, based on the rapid determination of differentially active spots, a variety of chromatographic techniques were used in combination to separate and purify the mutants new metabolites . Compound structure was identified based on physico-chemical properties and spectral data. Anti-tumor activity was determined by MTT assay. Results Six new metabolites of this mutant were isolated and identified from the mutant D2s4-1, namely, 1,9-dimethylphenazine (1), 2-hydroxyphenylacetamide (2) (3), daidzein (4), cyclo (4-hydroxy-proline-leucine) dipeptide (5) and uracil (6). Among them, 1, 3, 5 and 6 had certain antitumor activity, and the inhibitory rates of K562 cells were 33.3%, 16.3%, 33.3% and 21.1% at the concentration of 100μg / ml respectively. Conclusions Six new products of the antitumor activity mutant D2s4-1 were clarified, of which four were active products. Compound 1 is a new natural product, its anti-tumor activity is the first screening found. The active mutants obtained from the wild-type actinomycetes by chemical mutagen combined resistance screening technology can be used for screening new active products to expand the new strain of actinomycin-derived active strains.