Research Of Fibrinolytic Active Compound From Marine Fungi

Marine microorganisms, which live in special biotic environment, have been proven to be potential in producing novel bioactive substances. And a lot of marine bioactive substances produced by marine microbes have been discovered. With further study, the research and development of marine microorganisms attracts the focus of the world attention. In this paper, we had studied the isolation and structure determination of enhancing fibrinolytic active compounds from marine fibrinolytic fungi FG216 broth. Then fibrinolytic activity in vitro and acute toxicity of the isolated active compounds FGFC2 were studied. Finally, molecular biology means were used to identify fungal FG216.In the first chapter, secondary metabolites of marine microorganisms and their bioactivity which included antibiotic action, anti-tumor activity, anti-cardiovascular diseases, antioxidation, anti-inflammatory and enzyme inhibition since 2000 were summarized. Approximately 27% of Marine microorganisms can product antibacterial substances many of which were not exist in terrestrial. The seeking of new antitumor from marine microorganisms is a currently hot pot. Bioactive compounds with antitumor activity from marine microorganisms include nitrogenous compounds, lactones ketones, quinonoids and polysaccharides. In addition, Bioactive compounds with anti-oxidant activity, anti-inflammatory or enzyme inhibition from marine microorganisms have gained considerable attention. The present research indicated that more and more active compounds were founded from marine microorganism. However, little of that was used as the medcine which treated thrombosis.With the aim of looking for new drugs treated thrombosis, we had done some initial study that were described in the next chapters.In the second charpter, we have studied the isolation condition of enhancing fibrinolytic active compounds from the broth of FG216 by the extraction of methanol and semipreparative HPLC, respectively. The result revealled that we could isolate the target compound using the following chromatographic conditions, the column was developed at 40℃and the detection wavelength was 265nm and the velocity was 10mL/min. through twice refined (the first time used 50mM ammonium acetate in 70% aqueous MeOH as mobile phase, and the second time used 0.1 % formic acid in 40% acetonitrile), the enhancing fibrinolytic active compound FGFC2 was isolated. By the detection of purity, the compound FGFC2 can be used in the structure identification.In the third charpter, we have studied the structure character of compound FGFC2 by the spectrum of UV, IR, API-ESI-MS, 1~H-NMR, 13C-NMR, HMQC, 1~H–1~H COSY and HMBC. The compound FGFC2 belonged to intra-molecular symmetric compound. And we inferred it was statin metabolites and a novel compound after retrieving.In the fourth chapter, the effect of enhancing fibrinolytic of the active compound FGFC2 was measured with chromogenic assay. And the results show that in the reciprocal activation system of prourokinase and plasminogen,the activity was highest when the additive amount of fibrinolytic active compound was 10μg/mL. No pathological change or spiritual activity change was found in the experimental mouse by actual toxicity test. This shows safety and the toxicity was really small, which provided theoretic basis for the safety of clinical medication.In the fifth chapter, molecular biological method was used to identify the species of the marine fungi FG216. Phylogenetic tree based on ITS-5.8 S rDNA sequences showed that FG216 strain has a high homology with Stachybotrys longispora, but was not yet identifiable on species level of taxonomic classification.