| Malignant tumor is one of the diseases that seriously threaten human life and health.The development of new anti-tumor drugs with low toxicity and high efficiency has been a hot topic in the field of drug research,and the search for anti-tumor lead compounds from fungi is one of its main research directions.Polyporus umbellatus（Pers.）,a dried sclerotium,is a valuable traditional Chinese medicine,with various biological activities such as diuresis,immune regulation,anti-tumor,liver protection,and anti-oxidation,etc,and has a history of more than 2500 years in China.Modern pharmacology shows that the polysaccharides and steroidal compounds in Pers have stronger antitumor activities,which are usually used in clinical treatment with chemotherapy for lung cancer and viral hepatitis,and almost non-toxic side effects.Therefore,the separation and derivatization of polyporus umbellatus compounds is a very significant and necessary research.The main findings are as follows:The quality of Pers produced in Taibai Mountain in Shaanxi Province is the best.After smash,The 20 Kg Pers was ultrasonically extracted with 78%ethanol to obtain 655 g of ethanol extractives,and the yield of extract was 3.6%,which was significantly higher than the conventional reflux extraction（1.4%）and percolation extraction（1.98%）.Therefore,ultrasonic extraction is more advantageous than traditional methods such as reflux and percolation in the extraction of Chinese herbal medicine.The chemical components of ethanol extract were separated systematically by conventional column chromatography combined with modern preparation and separation techniques,and the chemical components of Pigs were analyzed in detail by liquid chromatography-mass spectrometry.Twenty-eight compounds were isolated and identified.Twenty-eight compounds includng seven steroids,four phenols,six nitrogen-containing compounds,and four carboxylic acids and their derivatives,of which cis-15-vaccenic acid methyl ester,and cis-15-vaccenic acid was isolated from the sclerotia for the first time.The crude polysaccharide was precipitated by water extract through ethanol,after hydrogen peroxide bleaching,sevage reagent to precipitate protein,dialysis,cellulose column chromatography,gel column chromatography,the molecular weights of water extract were 18706 Da and 22106 Da respectively,which are two homogeneous polysaccharides,and through the PMP derivatives were measured after they monosaccharide composition,xylose as main ingredients,contains a small amount of Fructose,Rhamnose and Mannose.The extracted alcohol extract,dichloromethane extract,n-butanol extract and Polyporus umbellatus polysaccharides were studied respectively for their antioxidant activity,DPPH free radical scavenging activity,antifungal activity and antibacterial activity.Polysaccharides showed strong anti-oxidation and scavenging free radical ability,but had no activity against fungi and bacteria.The cytotoxic activity of six isolated steroid compounds against human hepatocellular carcinoma cell line Hep G2,human cervical carcinoma cell line Hela,human renal clear cell cell 786-O and human proximal renal tubular epithelial cell HKC was tested,the results showed that the six steroid compounds showed strong cytotoxicity to the three kinds of cancer cells,and almost no cytotoxicity to the normal cells.The IC50 values of ZLW-5 on Hep G2 cells,Hela cells and 786-O cells were only 14.36μg/m L,21.90μg/m Ll and 46.56μg/m L.The strain is activated by PDA medium and mycelium is obtained by liquid liquid fermentation.After drying and grinding of the fermentation mycelia,19 compounds were obtained by ultrasonic extraction with ethanol,among which 13 compounds were identified.These 13 compounds included 5 steroids,2 nitrogenous compounds,2 phenolic compounds and 4 fatty acids and derivatives,and among them,nonadecanoic acid and methyl nonadecanoate were firstly isolated from the fermented mycelia.Synthesis of steroid derivatives:the ester derivatives of 22 ergosterols were synthesized with high yield of ergosterol and acyl chloride under mild conditions,of which8 compounds have not been reported.As a modifiable precursor compound that can be obtained from plant secondary metabolites,Dehydroepiandrosterone has been widely used in the study of steroid chemistry,In order to further expand the contents of steroidal chemistry and screen out highly active derivatives,we designed and synthesized a series of17-aza-androstene esters via hydroxyl protection,oximation,Beckman rearrangement and esterification using dehydroepianddroster one as starting material,Thirty-two of compounds have not been reported.The activities of 5k,5o and 5y were higher than those of positive control KMn O4,Moreover,brine shrimp letha:lity activity of these hybrids were tested and four effective compounds were screened out with LC50y ranging from5.34-9.81μg/m L and these results lay the foundation for further studies The IC50 values of CH-21 and CH-22 on Hep G2 cells,Hela cells and 786-O cells were only 24.68μg/m L,45.76μg/m L and 58.90μg/m L.respectively The IC50 values of 7g were only 19.88μg/m L,32.17μg/m L and 40.18μg/m L,so do research is a very meaningful and worthy of further research work.Six fatty acids were isolated from Pers for the first time,which enriched the number of compounds in Pers.High yield was derived not eight ergosterol esters and thirty-two 17-nitrogen impurity male steroid ester derivatives,select 5 of the cancer cells showed a greater toxicity steroidal compounds,all showed weaker toxicity to normal cells,for steroidal compounds derived and activity screening laid a certain foundation,... |
