Studies On The Bioactive Constituents Of Secondary Metabolites Of Two Marine-Derived Actinomycetes, 3295 And Z₂034a-5

We have undertaken the screening for novel cell cycle inhibitors and apoptosis inducers from the marine-derived actinomycetes, 3295 and Z2034a-5 by use of a mouse cdc2 mutant cell line, tsFT210. In anticipation of achieving the above objectives, marine-derived actinomycetes, 3295 and Z2034a-5 have been screened by using this model, the hexane, n-Butanol extracts of strain 3295 and EtoAc extract of strain Z2034a-5 showed bioactivities of inhibiting the cell cycle progression and cytoclasis of tsF210 cells. Thus, we selected them for further investigation.Two bioactive compounds 1-2 were isolated from the bioactive part, hexane and n-butanol extract of strain 3295, and six compounds 3-8 were isolated from the bioactive part, EtoAc extract of strain Z2034a-5, by repeated column chromatography on Sephadex LH-20, silica gel and RP18, followed by reverse-phase high performance liquid chromatragraphy. The structures of compounds 1-8 were elucidated mainly by use of spectroscopic method (UV, IR, MS, 1D-NMR, 2D-NMR) and according to their physicochemical properties: Phthalic acid dibutyl ester (1) , l-(l-methoxypropan-2-yl) 3-methyl 2-hydroxy isophthalate dimer (2) , methyl cinnamate (3), N-(4-hydro xyphenethyl) acetamide (4), cyclodipeptide (Tyr-Pro) (6), 5-(1-amino-l-oxo-3-phenylpropan-2-ylamino) -2,4- dihydroxypentanamide (7) , alkaloid-typed compound (8). The structure of compound (5) and (8) is still in affirming.Bioassay results indicated that compound (1) showed a strong apoptosis and cytoclasis of tsFT210 cells. It is the main antitumor active compound of hexane extract of strain 3295. Compound (2) could inhibit the cell cycle at different phase of tsFT210 cells and is the main antitumor active compound of n-butanol extract of strain 3295. Compound (3) show cell cycle inhibition and cytoclasis of tsFT210 cells;Compound (4) show an apoptosis and compound (5) show a tender cytoclasis of tsFT210 cells; Compound (7) show a strong bioactivity in inhibiting the cell progression at the G2/M phase of tsFT210 cells; Compounds (3) and (7) are the main antitumor active compounds of EtoAc extract of strain Z2034a-5. Compound (8) is novel in structure and possessed strong apoptosis and cytoclasis of tsFT210 cells.The antitumor activities of compounds 1-8 were assayed by SRB using tsFT210 cells. Compounds 1-8 exhibited antitumor activity. And compound (6) could stimulate the proliferations of tsFT210 cells tenderly at the concentration of 100 g /mL.The apoptosis activities of tsFT210 cells of compounds (1) and (4) in vitro were discovered for the first time; the inhibitory activity on the cell progression at the G2/M phase of tsFT210 cell of compound (3) in vitro was also discovered for the first time. Compounds (7) and (8) were discovered for the first time.Bioassay results indicated that we isolated the main antitumor active compounds of strain3295 and Z2034a-5 successfully which showed that this antitumor model and the method were exact and available in achieving bioactive components from marine microbes.