| Previous studies had indicated that the endophytic fungal metabolites may have homology with the host plant. In this paper, we studied the metabolites and biological activity of the endophytic fungus from the Cephalotaxus hainanensis Li [C. mannii Hook. f.] with anti-tumor activity. In order to discover new compounds with bioactivity, we selected a fungus and actinomycetes with good anti-tumor activity and genetic stability, based on the morphological evaluations of Pyricularia oryzae and Alternaria solani in bioassays. Their metabolites were investigated, respectively.The chemical constituents of the fermentation and/or mycelia of these three strains were studied by kinds of chromatographical techniques (such as silica gel chromatography, sephadex LH-20, PTLC and HPLC etc.). The structures of the compounds were elucidated on the basis of their spectroscopic and physico-chemical properties. The endophytic fungus L10 (Colletotrichum sp.) was isolated from the phloem of the Cephalotaxus hainanensis Li [C. mannii Hook. f.]. 33 compounds were isolated from the extracts of the fungus, elucidated as follow: 4- hydroxyphenyl ethanol- 2'- (R)- hydroxypropionate (L1), 4- hydroxyphenylethanol- 2'- (S)-hydroxypropionate (L2), 4- hydroxyphenylethanol- 2'- (S)- hydroxypropionate- 2"-(S)- hydroxypropionate (L3), 4- hydroxyphenylethanol- 2'- (S)- hydroxypropionate-2"- (R)- hydroxypropionate (L4), 4 - hydroxyphenylethanol pyroglutamic acid ester(L5), 4- hydroxyphenylethanol- 3- oxo- 2, 3- dihydro- 1H- pyrrolizine- 2- ester (L6), 4-hydroxyphenylacetic acid (L7), 4- hydroxyphenyl- methyl acetate (L8), 4-hydroxyphenyl ethanol (L9), 4- hydroxyphenyl ethylamine (L10), 6- (1' 2'- dimethyl-oxygen ring- 1 ')- 4- methoxy- 3- methyl- 2H- pyran- 2- one (L11), 6- (1-hydroxyethyl)- 4- methoxy -3 - methyl - 2H- pyran- 2- one (L12), Nectriapyrone(L13), Wermopyrone (L14), 4- methoxy- 3- methyl- 6- (2- methylene- 1- ene)- 2H-pyran- 2- one (L15), Harzialatone A (L16), Psoralen (L17), 5- hydroxymethyl furfural(L18), Isorubijervine- 3- O- ?- rhamnopyranosyl (1?4)-?- glucopyranoside (L19),Solanidine- 3- O- ?- rhamnopyranosyl (1?4)- ?- glucopyranoside (L20), 3 -indoleylethanol, 2, 2'- [1, 1'- methylene- bis- (1H- indole - 3, 1 - di- yl)] ethanol(L22), Pyroglutamic acid methyl ester (L23), Cyclo (Phe- Pro) (L24), Cyclo (Ala-Leu) (L25), Cyclo (Ala- Ile) (L26), Cyclo (Leu- Pro) (L27), Dibutyl phthalate (L28),Guanine (L29), Uracil (L30), Thymine (L31), Adenosine (L32), Glucose (L33). The fungus G20- 12 (Trichoderma atroviride) was isolated from the rhizosphere sediment of the Ceriops tagal, a kind of mangrove plant live in the intertidal zone of the South China Sea. 30 compounds were purified from the extracts and elucidated as follow respectively: Atroviridetide (G1),4'- (4, 5- dimethyl-1,3- diol- 2- yl)- methyl- phenol(G2), (3'- hydroxybutyl- 2'-yl)- 5- oxo- proline ester (G3), 3'- (3- oxo- cyclopentene-1- yl)- propionic acid n- butyl ester (G4), 3'- (3- oxo- cyclopentene- 1- yl)- propionic acid methyl ester (G5), Cyclo (Pro- Asp) butyl ester (G6), (E)- 3- (5- (hydroxymethyl)furan- 2- yl)- 2- benzeneylacrolein (G7), Pyroglutamic acid n- butyl ester (G8), 2, 3-dihydroxy benzoic acid (G9), ?- hydroxy acid anisole (G10), 4 - hydroxyphenyl ethanol acetate (G11), 4, 5- dihydroxy- hexanoic acid- ?- lactone (G12), 4 -hydroxybenzoic acid (G13), Succinic acid (G14), a- hydroxy- propionic acid (G15),Cyclo (Gly- Pro) (G16), Cyclo (Phe- Pro) (G17), Cyclo (Hyp- Pro) (G18), Cyclo(Leu- Pro) (G19), Cyclo (Val- Pro) (G20), Cyclo (Ile- Pro) (G21), Cyclo (Val- Tyr)(G22), Cyclo (Hyp- Phe) (G23), Cyclo (Leu- Hyp) (G24), Cyclo (Gly- Phe) (G25),Cyclo (Ala- Pro) (G26), Cyclo (Ala- Leu) (G27), Cyclo (Ala- Ile) (G28), Cyclo (Ala-Gly) (G29), Cyclo (Leu- Tyr) (G30). The actinomycetes TPF- 4 (Streptomyces tendae)was collected from Qingdao China. 9 compounds were isolated from the extracts and 8 of them were identified as follow: Emodin (T1), Chrysophanol (T2), Aloe- emodin(T3), 3- hydroxy- aloe- emodin (T4), 2, 5- dihydroxyphenylacetic acid (T5), Succinic acid (T6), Uracil (T7), Thymine (T8).Among them 14 compounds were new. Compound L1- L6 were new 4-hydroxyethyl esters, L11, L12 were new pyranone derivatives, L19 was new steroidal alkaloids, G2, G3 were new compounds which contain 2, 3- butanediol fragments, G1,G6 were new Cyclopeptides.Some of these compounds were tested for their cytotoxicity in vivo against two cell lines (human acute myeloid leukemia HL- 60 cells and human prostate cancer PC- 3 cells). Only compound L2 showed medium cytotoxicity against HL- 60 and PC- 3 cells. IC50 was 43.23 ?M and 63.09 ?M respectively. Other compounds tested showed none cytotoxicity in vivo against the two cell lines.In the research, we isolated the epimer L3, 4 successfully by three- phase system, providing the experimental basis for the future separation of these kinds of compounds. |
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