Preparation Of Dicarboxylic Acid Of Norcantharidin/Chitosan Salt And Studies On Its Anti-cancer Activity; Studies On Chemical Constituents In Bulgaria Inquinans And Its Antituberculous Activity

Aim: To prepare the dicarboxylic acid of norcantharidin/ chitosan salt, in order to reduce the toxicity of norcantharidin and enhance its anti-tumor activity. Methods: The chitosan samples with high degree of deacetylation, low molecular weight were prepared by an intermittent alkaline hydrolysis and H2O2 degradation, the dicarboxylic acid of norcantharidin was prepared by the hydrolysis. Subsequently, the negatively charged dicarboxylic acid of norcantharidin at acidic pH values allowed the formation of salt with the polycationic character of chitosan molecules. The acute toxicities of mouse treated with vena caudalis and lavage for the dicarboxylic acid of norcantharidin/ chitosan salt and the norcantharidin were observed. The median lethal dose (LD50) was calculated by Bliss software, furthermore the anti-tumor effect of the salt was investigated using the H22 hepatoma carcinoma model of mouse. Results: The initial experiment of anti-tumor activity validated that the salt has abated the toxicity of the norcantharidin and could inhibit the growth of tumors in mice. The biggest inhibitory rate was 44.38%, with a linear relationship between the effect of anti-tumor and concentrations. Conclusion: The dicarboxylic acid of NCTD/chitosan salt could enhance the anti- tumor activity, curative effect, and reduce the toxicity of of NCTD. Bulgaria inquinans belongs to the Ascomycotina family, is a fruiting body. It is widely distributed in temperate zone on northern hemisphere, and in china, it was in JiLin,HeBei,GanSu,SiChuan,YunNan province and so on. The fruiting body was used for treatment of H22 cancer of liver,U14 cervical cancer,S180 fleshy tumor and ehrlich ascite tumor cell, and also obviously cut down the Thrombosed length,dry weight of animal on blood stasis in vitro. In order to make further use the Chinese herbal resources and find active compounds, we have undertaken a systematic investigation on the chemical constituents of the Bulgaria inquinans and its antituberculous activities.Nine compounds were isolated from the ethanolic extracts of the fruiting body of Bulgaria inquinans and all of them were identified by using various modern spectrometric approaches, including IR, 1H-NMR, 13C-NMR, EI-MS, etc; the last two compounds were obtained from Bulgaria inquinans for the first time.Ergosterol peroxide was separated from Bulgaria inquinants; its analogue 7-dehydrodesmosterol also was oxygenized; the antituberculous acitivity of oxidative product were detected using MTT method; the result shows that the oxidative products have the antituberculous activities when the concentration of Ergosterol peroxide exceeded 20 ug/ml and the 7-dehydrodesmosterol peroxide exceeded 50ug/ml.