Study On Active Compounds With Specific Activation Of ER? Derived From Two Endophytic Fungi Annulohypoxylon Cf.Stygium(ARTCL-05)/Diaporthe Sp.(ARL-09)of Anoectochilus Roxburghii

Secondary metabolites of endophytic fungal is an important source of novel Compounds with novel structure and biological activity.Anoectochilus roxburghii(Wall.)Lindl is famous region drug in Fujian province,with many pharmacological activities.There are few reports on the secondary metabolites of endophytic fungi from Anoectochilus roxburghii(Wall.)Lindl,so the research on that is significant.Previously,three strains(Diaporthe sp.(ARL-09),Arthrinium arundinis(ARL-13)and Annulohypoxylon cf.stygium(ARTCL-05))with specific activation of ERa transcriptional activity were screened from 93 strains of endophytic fungi extracts by estrogen receptor(ERa)as target protein model.The main objective of this subject is to isolate and purify Compounds with specific activation of ERa transcriptional activity from the crude extract of strain Diaporthe sp.(ARL-09)and Annulohypoxylon cf.stygium(ARTCL-05).In this study,3 Compounds and 25 additional Compounds were isolated from the crude extracts of Annulohypoxylon cf.stygium(ARTCL-05)and Diaporthe sp.(ARL-09)by using TLC,RP-18 column chromatography,gel column Sephadex LH20 chromatography,normal phase silica gel column chromatography,HPLC,recrystallization and other chromatographic separation methods.The structure of 28 Compounds were established by physicochemical properties and various current spectroscopic methods(UV,IR,MS,NMR),mainly divided into isopentenyl indole alkaloids,nucleosides,coumarins,anthraquinone and its dimer,xanthenone and other types.Among them,Compound 1 of strain Diaporthe sp.(ARL-09)is a new Compound,and 26 Compounds are obtained for the first time from two genera.The isolated 28 Compounds were subjected to ERa transcriptional activity.Among them,Compounds 12,15 of strain Annulohypoxylon cf.stygium(ARTCL-05)and Compounds 4 of strain Diaporthe sp.(ARL-09)have specific transcriptional activation activity against ERa.The cytotoxic activity of the isolated 28 Compounds against 8 cell lines was tested.Among them,the Compound 2,12,13,17 of the strain Annulohypoxylon cf.stygium(ARTCL-05)and the Compound 11 of the strain Diaporthe sp.(ARL-09)had a good selective inhibitory activity against some cancer cells with a certain concentration-dependent.The Compounds with ERa transcriptional activity was found,which provided a scientific basis for the further study of the mechanism of specific action on ERa of the active Compound,the discussion of the structure-activity relationship,and the prospect of drug-making.