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目的阐明真菌萨氏曲霉(Aspergillus sydowi)D2-6具抗肿瘤活性的次级代谢产物。方法摇床发酵培养生产菌D 2-6,活性跟踪分离纯化D 2-6发酵液中的活性单体化合物,并根据理化性质和光谱分析(ESI-MS、UV、IR、NMR等)鉴定单体化合物结构;采用细胞形态镜检、MTT方法评价单体化合物对人类慢性髓性白血病K562细胞、人肺癌A549细胞、人肝癌BEL7402细胞、人白血病HL60细胞和小鼠白血病P388细胞的抑制活性。结果从真菌萨氏曲霉Aspergillus sydowiD 2-6发酵液中分离并鉴定了8个单体化合物,分别为5个二酮哌嗪类化合物fumitremorgin B(1)、dihydroxy-fumitremorgin C(2)、demethoxy-fumit-remorgin C(3)、fumitremorgin C(4)和gliotoxin(5),1个喹啉类化合物fumiquinazoline C(6),2个杂螺环-γ-内酰胺类化合物aza-spirofuran A(7)和azaspirofuran B(8)。化合物1~7的抗肿瘤活性检测结果显示,5对实验用肿瘤细胞的增殖抑制活性较强,对P388、A549、K562细胞的IC50分别为1.47、0.10、0.57 nmol.L-1,7对P388、A549、BEL7402的IC50分别为31.43、0.01、28.71μmol.L-1,6对A549细胞的IC50为42.10μmol.L-1,而1、2、3、4对实验用肿瘤细胞无明显抑制活性(IC50>100μmol.L-1)。结论真菌萨氏曲霉Aspergillus sydowiD 2-6的主要抗肿瘤活性次级代谢物是含硫二酮哌嗪类、杂螺环-γ-内酰胺类和喹啉类化合物,其中,含硫哌嗪类化合物5对P388、A549、K562细胞的生长具有较强抑制作用,杂螺环-γ-内酰胺类化合物7对A549细胞具有选择性增殖抑制活性。
Objective To elucidate the anti-tumor secondary metabolites of fungal Aspergillus sydowi D2-6. Methods The active metabolite D 2-6 was fermented and cultured in shaker. The active monomer compounds in D 2-6 fermentation broth were separated and purified by activity. The compounds were identified by their physico-chemical properties and spectral analysis (ESI-MS, UV, IR, NMR, etc.) The structure of the compounds was evaluated by MTT assay. The inhibitory activities of monomeric compounds on human chronic myeloid leukemia K562 cells, human lung cancer A549 cells, human hepatocellular carcinoma BEL7402 cells, human leukemia HL60 cells and mouse leukemia P388 cells were evaluated. Results Eight monomeric compounds were isolated and identified from the fungal Aspergillus sydowi D 2-6 fermentation broth. They were fumitremorgin B (1), dihydroxy-fumitremorgin C (2), demethoxy- fumitremorgin C (4) and gliotoxin (5), one quinoline compound fumiquinazoline C (6) and two aza-spirofuran A (7) And azaspirofuran B (8). The anti-tumor activity of compounds 1-7 showed that 5 had a strong inhibitory activity on the proliferation of experimental tumor cells with IC50 of 1.47,0.10,0.57 nmol.L-1,7 for P388, A549 and K562 cells respectively, , IC50 of A549 and BEL7402 were respectively 31.43,0.01,28.71μmol.L-1,6 had the IC50 of 42.10μmol.L-1 to A549 cells, while 1,2,3,4 had no significant inhibitory activity on the experimental tumor cells (IC50> 100 μmol.L-1). CONCLUSIONS The major anti-tumor activity secondary metabolites of the fungus Aspergillus sydowi D 2-6 are thiadione piperazines, heterospiro-γ-lactams and quinolines, among which the thiol piperazines Compound 5 has a strong inhibitory effect on the growth of P388, A549 and K562 cells, and the spiro spiro-γ-lactam compound 7 has a selective proliferation inhibitory activity on A549 cells.