Study On The Anti-inflammatory Constituents From Two Marine Organisms

Objective: In order to explore the lead compounds with efficiency, low toxicity and clinical application prospects, this paper studied on the chemical constituents and their antiinflammatory activities from the South China Sea soft coral Sinularia flexibilis and marine sponge Axinella sp..Methods: The soft coral Sinularia flexibilis was extracted with methanol, and the marine sponge Axinella sp. was extracted with ethanol. The dichloromethane dissolved fraction of soft coral and n-butanol dissolved fraction of sponge were separated, purified, and structure identified. Semi-preparative HPLC, reduced pressure column chromatography, atmospheric column chromatography, gel column chromatography, the preparation of thin layer chromatography and other separation methods were used to get the pure compounds from soft coral Sinularia flexibilis and marine sponge Axinella sp.. Based on the physical and chemical properties, combined with modern spectroscopic techniques, such as mass spectrometry(MS), nuclear magnetic resonance spectroscopy(1H NMR, 13 C NMR, 2D NMR) and other modern spectroscopic techniques, the reference related literature was used to determine the planar structure and the stereo configuration of the isolated compounds. MTT method was used in the macrophage(RAW264.7 cells) levels against inflammatory activity on the preliminary evaluation. Griess method was used to detect the inhibitory effect of LPS-induced NO production in RAW264.7 cells in noncytotoxic concentrations.ELISA was used to test the levels of TNF-? in the supernatant of LPS-induced RAW264.7 cells.Results: 11 compounds and 9 compounds were isolated from the soft coral Sinularia flexibilis and the marine sponge Axinella sp.. They were named and identified as: sinulaflexiolide M(1), sinulaflexiolide N(2), sinulaflexiolide O(3),sinulaflexiolide P(4), sinulaflexiolide Q(5), sinulaflexiolide R(6), sinulaflexiolide I(7), sinulaflexiolide L(8),(-)14-deoxycrassin(9), dihydrosinularin(10),24-methylenecholesterol(11), dibromophakellin(12), monobromophakellin(13),debromohymenialdisine(14), hymenialdisine(15), aldisin(16), 3-bromoaldisin(17),nicotinamide(18), thymine(19) and thymine(20) respectively. Compounds 1-6 were new compounds. Compounds 1-10 belonged to the cembranoid diterpens and compounds 12-17 were the pyrrole alkaloids. And compounds 18-20 were isolated from the Axinella sp. for the first time.Griess method was used to detect the inhibitory effect of LPS-induced NO production in RAW264.7 cells in noncytotoxic concentrations. The research showed that compounds 1, 2, 3, 4, 7, 8, 10, 12, 14, 15, 19 had certain inhibitory effect of NO,and the inhibition rate was 18.4%, 14.46%, 18.40%, 13.14%, 15.77%, 14.66%, 55.6%,10.84%, 11.21%, 13.46%, 9.52% respectively under 10 microns concentration.ELISA was used to test the levels of TNF-? in the supernatant of LPS-induced RAW264.7 cells. The result of the experiment exhibited that compounds 12 and 14 had weak inhibitory activity, and the inhibition rate was 5.08% and 0.70%respectively under 10 microns concentration.Conclusions: This study developed and enriched biodiversity and chemical diversity in the soft coral Sinularia flexibilis and marine sponge Axinella sp. colleted form the South China Sea to a certain extent. It also offered the chemotaxonomy of compound foundation and added the metabolites to the soft coral and marine sponge of the South China Sea. Of cuorse, it provided a scientific basis of marine natural products research and development as well.