Study On The Chemical Constituents Of Two Endophytic Fungi Isolated From Notopterygium Inchum Ting Ex H.T. Chang

Endophytic fungi of traditional Chinese medicine could produce not only a large number of secondary metabolites with novel structure and high biological activity,but also some endophytic fungi of traditional Chinese medicine could produce same or similar metabolites compared with their hosts.Thus,endophytic fungi of traditional Chinese medicine are attractive in traditional Chinese medicine research.Notopterygium inchum,as a common Chinese medicine,has the effects on relieving the exterior,dispelling cold,eliminating wind and dampness,relieving pain.With the purpose of searching secondary metabolites with novel structure and high biological activity,the chemical constituents of endophytic fungi from Notopterygium inchum were studied.This thesis consists three parts.In the first part,the chemical constituents on the n-butanol extracts of Myrothecium verrucaria,isolated from the root of Notopterygium inchum Ting ex H.T.Chang had been mentioned.During the second part,the chemical constituents on the secondary metabolites by epigenetic modification of an endophytic fungi Aspergillus fumigatus associated with Notopterygium inchum Ting ex H.T.Chang had been described.Myrothecium verrucaria,an endophytic fungus,was isolated from the fresh roots of Notopterygium inchum Ting ex H.T.Chang.14 compounds had been isolated from the ethyl acetate extract of this strain metabolites during our previous work.In this paper,further study on chemical constituents of n-butanol extracts of Myrothecium verrucaria metabolites,14 compounds were isolated.They were elucidated as cyclo-(Pro-Phe)(1),cyclo-(4-OH-Pro-Phe)(2),cyclo-(4-OH-Pro-Leu)(3),cyclo-(Pro-Ala)(4),cyclo-(4-methyl-Pro-9-propyl-Gly)(5),cyclo-(Pro-Gly)(6),cyclo-(Phe-Gly)(7),cyclo-(Leu-Leu)(8),N-acetyl tryptamine(9),N-phenylethyl-acetamide(10),N-(2-hydroxyphenethyl)acetamide(11),N-(4-hydroxyphenethyl)acetamide(12),uracil(13)and thymine(14)?Compounds 1~14were obtained from the strain Myrothecium verrucaria for the first time.The gene of endophytic fungi of traditional Chinese medicine,which couldproduce secondary metabolites,is high diversity.But,under normal laboratory culture condition,many genes of endophytic fungi have been silent or expressed at low level.In order to improve the expression level of silence gene,to produce more new bioactive compounds.Strains were fermented with medium adding epigenetic modifier 5-azacytidine(5-AC)or suberoylanilide hydroxamic acid(SAHA),and the EtOAc extracts were evaluated with chemical composition analysis method(HPLC).Thus,the endophytic fungus Aspergillus fumigatus of Notopterygium inchum Ting ex H.T.Chang had been choosed after adding SAHA in medium,and the chemical constituents of ethyl acetate extracts of Aspergillus fumigatus metabolites were studied.Finally,8 compounds were isolated and their structures were elucidated as5 a,6-dehydro-3-dehydroxymethylbisdethio-3,10a-bis(methylthio)glitoxin(1),bis(dethio)bis-(methylthio)gliotoxin(2),gliotoxin(3),verruculogen(4),12,13-dihydroxyfumitremorgin C(5),acetanilide(6),AM6898A(7),pseurotin A(8).Compounds 1 was obtained from natural products for the first time,and compounds 6and 7 were obtained from Aspergillus fumigatus for the first time.