Studies On The Secondary Metabolites And Bioactivities Of Three Endophytic Fungus From Sponge Phakellia Fusca Thiele

The Traditional Chinese Medicine(TCM) Freshwater Sponge, derived from dry group body of Spongilla fragilis(Leidy), was mainly distributed in Jiangsu, Henan provinces in China. It tastes sweet and its function is warm, which can be used as medicine to enrich the kidney and strengthening Yang-qi, as well as treat diseases such as impotence, spermatorrhea, gonorrhea, urinary incontinence, scrotal moisture and itch. Freshwater Sponge have been used for a long history in China and it was recorded by many ancient medicine books, such as Gleaning Herb, Compendium of Materia Medica, Chinese Materia Medica. It contains a wide array of structurally diverse and pharmacologically active secondary metabolites, and its endophytic fungus as a kind of microbial resource has become an important source of new drugs and leading compounds research and development. In order to elucidate the material basis of Freshwater Sponge effective constituent, to explore a new idea for the development and utilization of Freshwater Sponge, and to look for new active leading compounds, this thesis investigated the secondary metabolites of strain Arthrinium arundinis ZSDS1-F3, Glimastix sp. ZSDS1-F7 and ZSDS1-F11, which are endophytic fungus isolated from a closely related species of the Freshwater Sponge--Phakellia fusca Thiele, collected from the Xisha Islands of China, and the endophytic fungus was identificated by colony morphology and 18 S r RNA gene sequence analysis.According to the marine natural products research method, the selected endophytic fungus strains were incubated in the optimized fermentation culture medium, following the HPLC fingerprint analysis and preliminary bioactive screening. The fermented products were purified by means of chromatography over silica gel column, ODS column, Sephadex LH-20 and semi-preparative HPLC. The structures were confirmed based on their physical and chemical properties, such as modern spectroscopic analysis(NMR, MS, IR, UV,CD, X-ray diffraction)and comparison with the relevant literature. Twenty nine compounds were identified, including a new natural product A1. Six compounds were isolated from the endophytic fungus Arthrinium arundinis ZSDS1-F3 derived from the sponge Phakellia fusca Thiele. These compounds were identified as: phenethyl 5-hydroxy-4-oxohexanoate(A1), trans-(3R,4S)-(-)-4-hydroxymellein(A2), cis-(3R,4R)-(-)-4-hydroxymellein(A3), decarboxycitrinone(A4), altechromone A(A5), R-(-)-5-hydroxymellein(A6). Fifteen compounds were isolated from the endophytic fungus Glimastix sp. ZSDS1-F7 derived from the sponge Phakelliafusca Thiele. These compounds were identified as: purpurogemutantin(G1), macrophorin A(G2), 4'-oxomacrophorin(G3), 7-deacetoxyyanuthone A(G4), 2,3-hydro-deacetoxyyanuthone A(G5), 22-deacetylyanuthone A(G6), anicequol(G7), roquefortine C(G8),(16S)-hydroxyroquefortine C(G9),(16R)-hydroxyroquefortine C(G10), dihydro- resorcylide(G11), nectriapyrone(G12), methyl linoleate(G13), phospholipase A2(G14), methyl elaidate(G15). Eight compounds were isolated from the endophytic fungus ZSDS1-F11 derived from the sponge Phakellia fusca Thiele.These compounds were identified as: Tajixanthone hydrate(K1),S-curvularin(K2), 1H-indole-3- carboxal dehyde(K3),cyclo(L-Val-L-Ala)(K4),cyclo(L-Ala-L-Ile)(K5),cyclo(L-Ala-L-Leu)(K6), cyclo(L-Tyr-L-Pro)(K7), cyclo(L-Trp-L-Phe)(K8).On the basis of the structures of compounds and reference, these compounds were tested the preliminary bioactivity. Among these compounds, compounds A2, A3, G1-G3, G5-G7 showed significant invitro cytotoxicities against the K562, MCF-7, Hela, DU145, U937, H1975, SGC-7901, A549, MOLT-4 and HL60 cell lines with IC50 values ranging from 0.19 to 35.4 ?M. Compounds G2-G4 exhibited antitubercular activity with IC50 values of 22.1, 2.44 and 17.5 ?M, respectively. Furthermore, compound G7 had anti-enterovirus 71 activity with MIC value of 17.8 ?M. The research above further enriched the natural products compound library of marine drug screening, and provided the active template compounds for the drug research of antitumor, antitubercular and anti-EV71 virus, as well as the basic data for the secondary metabolites research of the sponge-derived fungus and certain ideas for the development and utilization of traditional Chinese medicine Freshwater Sponge.