The Investigation On The Secondary Metabolites Of Penicillium Sp.,a Fungus Obtained From The Rhizosphere Of Panax Notoginseng

Rhizosphere microorganism is likely to contain many new bioactive secondary metabolites due to the complex relationship among the plant,rhizosphere and microorganism,so rhizosphere microorganism has become a new resourse to look for new natural bioactive lead conpounds.In this thesis,an investigation was carried out on the secondary metabolites of a strain of Penicillium sp.(NO.SYP-F-7919),a fungus obtained from the rhizosphere of Panax notoginseng collected from the Yunnan province of China.51 compounds were isolated from the culture broth and the solid fermentation culture of SYP-F-7919 through various chromatographic techniques,such as silica gel column chomatography(CC),Sephadex LH-20 CC,ODS CC and preparative high performance liquid chromatography(p-HPLC).On the basis of physico-chemical property,spectroscopic analyses,calculated electronic circular dichroism(ECD),chemical derivatization and single crystal X-ray diffraction analysis.The structures of compounds were identified as penicimenolide A(1*),penicimenolide B(2*),penicimenolide C(3*),penicimenolide D(4*),penicimenolide E(5*),penicimenolide G(6*).cis-resorcylide(7),dihydroresorcylide(8),(7S)-methoxyresorcylide(9),(7R)-methoxyresorcylide(10),(7R)-hydroxydihydroresorcylide(11),(4S)-hydroxydihydroresorcylide(12),penicimenolide F(13*),orthosporin(14),(E)-6,8-dihydroxy-3-(6-oxo-1en-1-heptyl)-1H-isochromen-1-one(15),2’,3’-dihydrosorbicillin(16),sorbicillin(17),trichodimerol(18),4-(2-formyl-5-methoxymethylpyrrol-l-yl)butyric acid methyl ester(19),N-(2aminophenyl)urea(20),p-hydroxyphenylacetic acid methyl ester(21),protocatechuic acid(22),trans-4-methyl-4-hydroxycrotonic acid(23),penicimenolide H(24*),penicimenolide I(25*),(75)hydroxydihydroresorcylide(26).O-desmethylgreensporone C(27),penicimenolide J(28*),orsellinic acid(29),penicisocoumarin A(30*),penicisocoumarin B(31*),penicisocoumarin C(32*),penicillsterone A(33*).(22E,24R)-5α,8α-ergosta-6,9(11),22E-trien-3β-ol(34),5,8-epidioxy5α,8α-ergosta-6,22E-dien-3β-ol(35),(22E,24R)-5α,8α-epidioxy-23-methylergosta-6,22-dien-3β-ol(36).penicialide A(37*),7-hydroxy-5-methoxy-4-methyl-6-(3-methyl-4-oxopentyl)-phthalan-l-one(38),methyl mycophenolate(39),6-(3-carboxybutyl)-7-hydroxy-5-methoxy-4-methylphthalan-1one(40),2ω-dihydroxyemodin(41),citreorosein(42),emodic acid(43),2-chloroemodic acid(44),9-octadecenoic acid-2’,3’-dihydroxy propyl ester(45),2.3-dihydro-6,7-dihydroxv-4H-1benzopyran-4-one(46).5,7-dihydroxy-isobenzofuran(47),3,5-dihydroxytoluene(48),catechol(49),striatisporolide A(50),monoethyl fumarate(51).15 compounds(1-6,13,24,25,28,30-33,37)are new compounds and 4 compounds(15,38,39,50)are new natural products.In addition,8 compounds(1,2,5,7,8,10,13,18)were isolated from both the culture broth and the solid fermentation culture of SYP-F-7919.13 compounds(1-5,7-12,16 and 18)isolated from the culture broth of SYP-F-7919 were evaluated for their cytotoxic activity against six human tumor cell lines,including U937,MCF-7,A549,SH-SY5Y,HepG2 and SW480.Compounds 2-4,7 and 18 exhibited cytotoxicity against the U937,MCF-7,SH-SY5Y and SW480 tumour cell lines.The substitution of an acetvloxy or 2hydroxypropionyloxy group at C-7 significantly increased the cytotoxic activity of the resorcylic acid lactone derivatives.Subsequently,the possible mechanism of compound 2 against MCF-7 cells was preliminarily investigated by in silico analysis and experimental validation,indicating compound 2 may act as a potential MEK/ERK inhibitor.Moreover,proteomics analysis was performed to explore compound 2-regulated concrete mechanism underlying MEK/ERK pathway,which is still need further study in the future.In addition,30 compounds(1-5,7-12,24,27-29,31-39,41-46)isolated from the culture broth and the solid fermentation culture of SYP-F-7919 were assayed for their inhibitory effect on NO production in LPS-activated murine macrophages(RAW 264.7).Compounds 2-4,7,27-29,31-32,37 and 41 exhibited inhibitory effect equivalent to or higher than positive control hydrocortisone.and compounds 5,8,10,11 and 24 showed weak inhibitory effect.