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新制癌菌素是1965年发现的烯二炔类抗肿瘤抗生素,聚(苯乙烯-马来酸)-NCS偶联物在日本主要用于临床的肝癌介入治疗,目前该药物在国内还没有生产。为了制备新制癌菌素。新制癌菌素产生菌Streptomyces carcino-staticus ATCC15944被在28℃、250 r/min条件下液体发酵,振荡培养72 h。发酵液经预处理后,离心后取上清液经硫酸铵分级沉淀、纯水膜透析分离纯化得到组分SIPI-ZX-1。HPLC分析表明产物纯度大于90%。SDS-PAGE和质谱数据显示,该蛋白质相对分子质量约为11 ku。该产物的抗人肝癌细胞的体外活性被测定。该结果与新制癌菌素的分子质量和抗肿瘤活性数值基本一致。本制备方法简捷,成本较低,本工作为进一步研究新制癌菌素的制备工艺奠定了基础。
New neocarzinum is an enediyne antitumor antibiotic discovered in 1965. Poly (styrene-maleic acid) -NCS conjugate is mainly used in clinical interventional treatment of liver cancer in Japan. At present, the drug has not been produced domestically . In order to prepare neocarzinones. The neocarcinoma-producing strain Streptomyces carcino-staticus ATCC15944 was fermented at 28 ° C and 250 r / min and shaken for 72 h. After pretreatment of the fermentation broth, the supernatant was centrifuged and fractionated by ammonium sulfate, and the pure SIPI-ZX-1 was isolated and purified by membrane dialysis. HPLC analysis showed product purity greater than 90%. SDS-PAGE and mass spectrometry data show that the molecular weight of the protein is about 11 ku. The in vitro activity of this product against human hepatoma cells was determined. The results are consistent with the neocarticin molecular weight and anti-tumor activity values. The preparation method is simple and the cost is low. This work lays the foundation for further research on the preparation technology of neocarzinomics.