Studies On The Bioactive Constituents Of Secondary Metabolites Of Marine-Derived Fungus KLEB-07, H2-02 And WZ4-F22

We have undertaken a piece of research on the metabolites of marine-derived fungus, focused on apoptosis inducing and/or cell cycle inhibitory components, by use of bioassay on a mouse cdc2 mutant cell line, tsFT210. At the first step of this work, we selectively isolate 183 fungus from 24 sea-mud and screen 14 strains which have obvious antitumor activity .We select three marine fungus, KLEB-07,H2-02 and WZ4-F22, and study their cultivated conditions. In anticipation of achieving the bioactive components, marine-derived fungus, KLEB-07,H2-02 and WZ4-F22 have been screened by using this model, the EtOAc extracts of this three strains show bioactivities of apoptosis inducer and cytoclasis of tsF210 cells. Thus, we select them for further investigation.23 compounds are isolated from the bioactive part, EtOAc extract of the three strains, by repeated column chromatography on silica gel, Sephadex LH-20 and RP-18, followed by reverse-phase high performance liquid chromatragraphy. The structures of 19 compounds are elucidated mainly by use of spectroscopic method (UV, IR, MS, ID NMR, 2D NMR) and according to their physicochemical properties: Ergosterol(1), Ergosterol peroxide(2), 24(R)-ergosta -6 β -methoxyl-3 β ,5 α -dihydro-xy-7,22(E)-diene(3), 24(R)-ergosta-3 β ,5 α ,9 α -trihydroxy-7,22-diene-6-one(4), 2',3'-dihydrosorbicillin(5), Sorbicillin(6), 3-hex -4-enoyl-2,5-dimethyl-(l,4)Benzoquinone(7),Dihydrotrichodimerol(8), Trichodimerol (9),2-Nitro-benzene-l,3-diamine(10),2,6-Dinitrophenyl-amine(ll), 1 H-Indole-3-carboxylic acid(l2),Acetic acid3-hydroxy-5-hydroxymethylphenyl ester(13), Gliotoxin(14), Gliocladin(15),cyclo-(Trp-Ala)(16), cycol(Gly-Pro)(17), methyl-octadec-enoic acid(18),Hexadeca-6,9-dienoic acid 2,3-dihydroxypropyester(19).Bioassay results using FCM and SRB indicat that compounds 1,15,18 show a strong bioactivity in cytotoxic activity on tsFT210 cell. They are the main anticancer active compounds of KLEB-07. Compounds 2,4,5,6,7,8,9 show a strong bioactivity in apoptosis inducing and cytotoxic activity on tsFT210 cell. They are the main anticancer active compounds of H2-02. And compounds 3,14,19 show a strong bioactivity in apoptosis inducing and cytotoxic activity on tsFT210 cell. They are the main anticancer active compounds of WZ4-F22.Bioassay results indicat that we have isolated the main antitumor active compounds of strain KLEB-07,H2-02 and WZ4-F22 successfully which show that this antitumor model and the method are exact and available in achieving bioactive components from marine microbes.