Study On Chemical Constituents Of Two Traditional Chinese Medicine Related Bacteria

This thesis consists of three chapters.The first two chapters respectively detail the fermentation,isolation and identification of the fungus Penicillium crustosum isolated from the endophytic bacteria of Cordyceps sinensis and the epiphytic Streptomyces sp.KIB3133 from the rhizosphere soil of the traditional Chinese medicine tea tree,and the biological activities of these compounds were studied.The third chapter is a review of the research progress of natural sources of nanaomycins.The extract which obtained by solvent extraction was separated and purified by silica gel column chromatography,MCI,gel column chromatography and high performance liquid chromatography(HPLC),and was subjected to ESIMS,1D-NMR,2D-NMR(1H-1H COSY,HSQC),HMBC,ROESY,UV,IR,CD)and other spectroscopy methods for structural analysis of these compounds;the absolute configuration of the new compounds were determined by ECD calculation,Mosher ester NMR analysis and specific optical rotation;using chemical synthesis speculate the biosynthesis pathway of dimeric pyranonaphthoquinone from Streptomyces sp.KIB3133;the isolated compounds was subjected to zebrafish test,cytotoxic activity test,anti-HIV-1 activity test and antibacterial activity test.9 compounds were isolated from the endophytic fungus Penicillium crustosum.Three of the new compounds are(3S,6R)-3-(4-((S)-2,3-dihydroxy-3-methylbutoxy)benzyl)-1,4-dimethyl-3,6-bis(methylthio)piperazine-2,5-dione(1),(3S,6S)-3-(4-((S)-2-hydroxy-3-methoxy-3-methylbutoxy)benzyl)-1,4-dimethyl-3,6-bis(methylthio)piper azine-2,5-dione(2),(3S,6R)-3-(4-((S)-2-hydroxy-3-methoxy-3-methylbutoxy)benzyl)-1,4-dimethyl-3,6-bis(methylthio)piperazine-2,5-dione(3);6 known compounds:(-)-cis-bis(methylthio)silvatin(4),(S)-1,4-dimethyl-6-(4-((3-methylbut-2-en-1-)Yl)oxy)benzyl)-6-(methylthio)piperazine-2,3,5-trione(5),(3R,6S)-6-(para-hydroxy benzyl)-1,4-dimethyl-3,6-bis(methylthio)piperazine-2,5-dione(6),6-Acetyl-2,5-dihydroxy-2-(2-hydroxypropyl)-3,8-dimethylchroman(7),4-hydroxyphenylethanol(8),4-(2,4,7-trioxabicyclo [4.1.0] heptan-3-yl)phenol(9).Compound 1-5 was tested for gastrointestinal motility activity,and compound 4-5 were found to act on the cholinergic nervous system to promote gastrointestinal motility in zebrafish.Streptomyces sp.KIB3133 from the rhizosphere soil of tea tree was isolated 22 compounds,including 8 new S-bridged pyranthenequinones were naquihexcin C-J(1-8),naquihexcin K(10),OM-173αE(11),these compounds are all derived from Streptomyces;naquihexcin A(18),OM-173αE(20),nanaomycin βE(21),2754B(22),tectone(23),methyl phenylacetate(24),2754A(25),Benzeneacetic acid(26),4αA,10αB-dihydroxynanaomycinβA(27),Cyclo(L-Pro-L-Leu)(28),herbicidins A(29),chrysophanol glucuronide(30)and OM-173αB(31)are known compounds.Cytotoxic activity tests were performed on some new S-bridged pyranthroquinone compounds,targeting MCF-7(breast cancer cells)and HL-60(leukemia cells),with IC50 values ranging from 1.41 to 16.08 μM.When screening some new compounds for anti-HIV-1 activity,naquihexcin E was found to have weak activity with an EC50 value of 2.83 ± 1.15 μM and a TI value(CC50/EC50)of 22.51.Screening for antifungal and antibacterial activity of known compounds showed that some of these have good activity.In summary,A total of 31 compounds were isolated and identified from the two strains,of which 12 compounds were new.The structure of these compounds mainly were thiodiketopiperazine,pyranthenequinone,cyclic dipeptide,alkaloid and so on.