海绵共附生真菌Nigrospora sp.的次级代谢产物及PD-1/PD-L1抑制活性研究  

Secondary metabolites and PD-1/PD-L1 inhibitory activity from the sponge-associated fungi Nigrospora sp.

作　　者：刘明香 许聪聪 李冉[2] 吴文惠[1] 张文[2] LIU Ming-xiang;XU Cong-cong;LI Ran;WU Wen-hui;ZHANG Wen(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China;School of Pharmacy,Second Military Medical University,Shanghai 200433,China)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]中国人民解放军第二军医大学药学院,上海200433

出　　处：《中国海洋药物》2020年第3期1-6,共6页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(81820108030)资助。

摘　　要：目的研究海绵Callyspongia sp.共附生真菌Nigrospora sp.中化学成分以及生物活性。方法利用正相硅胶柱层析、Sephadex LH-20凝胶柱层析和半制备型反相高效液相色谱(RPLC)等方法对该菌株发酵产物的乙酸乙酯提取物进行分离纯化;通过核磁共振谱(NMR)、质谱(MS)等技术并结合文献报道数据分析鉴定化合物结构;采用均相时间分辨荧光(HTRF)法检测化合物的PD-1/PD-L1结合抑制活性。结果分离得到7个已知化合物,分别为punctaporonin D(1),punctaporonin E(2),punctaporonin F(3),desuccinoylpunctaporonin C(4),(20S,22E,24R)-麦角甾-7,22-二烯-3β,5α,6β-三醇(5),(22E,24R)-麦角甾-3β-羟基-5α,9α-环氧-7,22-二烯-6-酮(6),麦角甾-7,22-二烯-6β-甲氧基-3β,5α-二醇(7)。在体外生物活性检测中,化合物6显示出PD-1/PD-L1结合抑制活性,其IC50值为22.5μmol/L。结论化合物4为新天然产物,化合物1~3和化合物6为首次从该属真菌中分离得到。化合物6显示PD-1/PD-L1结合抑制活性。Objective To investigate bioactive secondary metabolites of fungus Nigrospora sp.associated with the sponge Callyspongia sp..Methods The ethyl acetate extract of the fermentation products of the strain was isolated and purified by normal phase silica gel column chromatography,gel Sephadex LH-20 and semipreparative reversed phase liquid chromatography(RPLC).The structures of the compounds were identified by modern spectroscopic technologies including nuclear magnetic resonance(NMR)and mass spectrometry(MS).The PD-1/PD-L1 binding inhibitory activity of the compounds were detected using homogenegous time resolved fluorescence(HTRF).Results Seven known compounds were isolated and identified as punctaporonin D(1),punctaporonin E(2),punctaporonin F(3),desuccinoylpunctaporonin C(4),(20 S,22 E,24 R)-ergosta-7,22-dien-3β,5α,6β-triol(5),3β-hydroxy-5α,9α-epoxy-(22 E,24 R)-ergosta-7,22-dien-6-one(6),ergost-7,22-dien-6β-methoxy-3β,5α-diol(7).Compound 6 showed PD-1/PD-L1 binding inhibitory activity with IC50 of 22.5μmol/L.Conclusion Compound 4 was reported as a new natural product while compounds 1~3 and 6 were reported from the Nigrospora sp.for the first time.Compound 6 showed a PD-1/PD-L1 binding inhibitory activity in in vitro biotest.

关 键 词：海绵共附生真菌 黑孢霉属 次生代谢产物 PD-1/PD-L1抑制活性 

分 类 号：R914[医药卫生—药物化学]