Biological Evaluation And Derivative Synthesis Of A Marine Bisindole Fibrinolytic Compound FGF1

Marine microorganism is an important resource,which provides rich diversity of natural products that pave the way for scientists to discover bioactive compound with antioxidant,anti-inflammatory,anticancer,inhibiting enzyme,antifungal,antibacterial,antialgal,antilarval activities,etc.At present,cardiovascular diseases(CVs)are the main cause of death among non-communicable diseases.According to the World Health Organization's reports,CVs were responsible for 17.5 million deaths in 2012,accounting for 31% of worldwide deaths.Moreover,thrombosis is one of the most common causes of CVs.While blood clots form in the blood vessels,blood circulation can be suffocated naturally.Due to ischemia and hypoxia,lesions of function and organism can be induced by embolization of tissues or organs.At the same time,thrombosis diseases endanger human physical and mental health seriously.Furthermore,it brings a heavy economic burden to patients' family and society.Consequently,the antithrombotic drugs with highly level of therapy effects,low risk of toxicity and economy should be developed.In previous studies,we isolated FGFC1 from Stachybotrys longispora FG216 with significant fibrinolytic activity in vitro and in vivo.The results showed that 0.1—0.4mmol/L of FGFC1 could stimulate generation of plasmin activity(increased by2.05—11.44 folds)by measuring Glu-plasminogen and Lys-plasminogen activation in vitro.As a part of our continuing work to search for bioactive leading molecules from marine microorganism,ethyl acetate fraction of organic extract of the train S.longispora FG216 exhibited fibrinolytic activity in our primary caused isolation of a new isoindolone,FGFC2(FGFC2,Fungi fibrinolytic compound 2),together with two known compounds,LL-Z1272? and ergosterol.The structure of FGFC2 was elucidated by the spectral analysis of 1D(1H,13C)NMR,2D(COSY,HSQC,and HMBC)and ESI-MS.Three compounds were evaluated for their fibrinolytic activities in vitro.Compared to FGFC1(EC50=47 ?M)as a reference drug,compound 1 and ergosterol(3)presented moderate fibrinolytic activities in vitro with EC50 value of 108.16 and 156.30?M,respectively.LL-Z1272?(2)had no fibrinolytic activity.And FGFC1 isolated with significant promote fibrinolytic activity,but has still some deficiencies in medicine such as water soluble and unstable poor structure.So FGFC1 group should be modified to change its deficiency or improve activity.In this thesis,phenolic hydroxyl groups of FGFC1 on the benzene ring is going to have astructure modification,aiming to change the fat-soluble for a preliminary experiment.The reaction is using a bromide with phenolic hydroxyl and carboxyl,in which phenolic hydroxyl parts generate ether and carboxylic parts generate ester.At last,there are four derivatives received.Among them,F-3 has the highest rate and promote the best fibrinolytic activity(EC50 = 42.29 ?M),the experimental results are useful for follow-up studies on the FGFC1 structure modification,and other parts of the groups of experiments are under way.In recent years,apoptosis researches have achieved good results,a growing studies show that there is a close relationship between apoptosis and a variety of thrombus disease,so we will do some cell experiment of the above three kinds of compounds isolated and structure modification of four types of derivatives and FGFC1.In this experiment,Hela cells are adopted which are more easily survived.The cytotoxicity,apoptosis factor Fas/Apo-1 and inflammatory factor IL-1 are tested.In the result,it shows that 8 compounds have certain cytotoxicity and certain inhibitory effect to the Hela cell,FGFC1 exhibits the strongest inhibitory effect.And in the experiment of Fas/Apo-1,in addition to which LL-Z1272 group has the fairly level as blank control group,the other seven kinds of compounds have all effected on Hela cell and have the apoptosis effect.Among them,FGFC1 with the strongest fibrinolytic activity exhibits the strongest effect on promoting apoptosis.With the increase of the concentration,the effect on promoting apoptosis of Hela also increases.The remaining six compounds on Hela cells also have different effects on promoting apoptosis,and apoptosis effect of the derivative F-1 has the similar effect as the effect of FGFC1,overall the increasing cytotoxicity is changing with the increasing concentration.In the experiment of IL-1,the FGFC1 group,FGFC2 group and the high concentration of four derivative groups exhibit higher data than the blank control group data,but according to the statistical significance,p>0.1 data was not significant,therefore it cannot explain causeinflammatory reaction.While FGFC5 group and FGFC6 group data are lower than the blank control group,p<0.1,there is significant difference,that means they will not cause inflammatory reaction.Studies have shown that the concentration of plasminogen activator has a certain relationship with cancer cell apoptosis,but the anticancer mechanism of these compounds still need further study.