真菌萨氏曲霉D2-6抗肿瘤活性代谢产物

Antitumor Metabolites from Fungus Aspergillus sydowi D2-6

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D2-6，活性跟踪分离纯化D2-6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HLS0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AspergillussydowiD2-6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fumitremorginB（1）、dihydroxy—fumitremorginc（2）、demethoxy—fumitremorginC（3）、fumitremorginc（4）和gliotoxin（5），1个喹啉类化合物fumiquinazolineC（6），2个杂螺环-γ-内酰胺类化合物aza-spirofuranA（7）和azaspirofuranB（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、0．57nmol·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 sydowi D2-6的主要抗肿瘤活性次级代谢物是合硫二酮哌嗪类、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 investigate the antitumor metabohtes from fungus Aspergillus sydowi D 2-6. METHODS The strain D 2-6 was fermented in a rotary shaker and the bioactive metabolites in the fermentation broth were isolated through a bioassay-guided separation procedure. Structures of the bioactive compounds were determined on the basis of physicochemiCal and spectroscopic data. The antitumor activity of the compounds against various tumor cell lines（ K562, A549, BELT402, HL60 and P388） in vitro was assayed by MTT method, accompanied with cell morphological observation under light microscope. RESULTS Eight compounds were isolated from the fermentation broth of Aspergillus sydowi D 2-6, and identified as five diketopiperazine derivatives including fumitremorgin B （ 1 ）, dihydroxy-fumitremorgin C （2）, demethoxy-fumitremorgin C （3）, fumitremorgin C （4）, gliotoxin （ 5）, one quinazoline derivative fumiquinazoline C （6）, two hetero-spirocyclic γ-lactam derivatives azaspirofuran A（7） and azaspirofuran B （8）. Antitumor activity assay displayed that compound 5 had the strongest inhibitory activity on test cell lines among the detected compounds. Compound 5 inhibited the growth of P388, A549 and K562 cells with IC50 of 1.47, 0. 10 and 0. 57 nmol · L^-1 , respectively. Compound 7 inhibited the proliferation of P388, A549 and BEL7402 cells with IC50 of 31.43, 0. 01 and 28. 71 μmol·L^-1 , respectively. Compound 6 inhibited the growth of K549 cells with IC50 of 42. 10 μmol·L^-1. CONCLUSION The predominant antitumor metabolites from Aspergillus sydowi D 2-6 were sulphureous diketopiperazine, hetero-spirocyclic γ-lactam and quinazoline derivatives, among which compound 5 showed strong inhibition on P388, A549 and K562 cells, while compound 7 exhibited specific inhibitory activity on A549 cells.