Qualitative Analysis Of Lytta Caraganae Pallas、Content Determination And Antitumor Activity Investigation Of Cantharidin And Its Derivatives

Insect medicine is an important part of Chinese traditional medicine.Cantharides,theclassical traditional insect medicine,mainly extracted from Mylabrisphalerata Pallas or Mylabriscichorii Linnaeus.With the increasing requirements of Cantharides,discovering new resource of these compundsis highly valuable,but also a taugh challenge.Based on these considerations,in this paper,some common butrarely studied Chloranthus Viridis were selected as our research objects,the detailed research works were depicted as follows: 1.Preliminary Investigation on the Varieties of Cantharides in PharmacopoeiaA survey was carried out in conjunction with the second national survey on terrestrial wildlife conducted by the State Forestry Administration.The main purpose was to use this opportunity to train graduate students to collect experimental samples.The results showed that the distribution of Melalea magna and Mela magna were investigated in 12 areas of Xingye County and Nanning City,Yulin City and the number was seen to be large.2.Extraction of cantharidin from Lytta caraganae PallasIn this part,the two varieties of Cyanine and Pharmacopoeia were compared.In the Pharmacopoeia 2015 edition,the methods for extracting Daphne Grandis and Daphne Cyclophyllawith chloroform were used.In this paper,the method of ultrasonic extraction was used in the pharmacopoeia,but the solvent was acetone.In order to determine the difference of extraction rate between the two solvents,the efficiency of cantharidinsextraction from Melittinusviridis was determined.In this experiment,two varieties of Chloranthusviridis and Pharmacopoeia were compared according to the determination method of Pharmacopoeia 2015.The peak value,peak area and extraction rate of these were compared.The results showed that the content of cantharidin was 0.932(1.014 for Cantharidins).The content of cantharidin was 0.9,and the content of cantharidin in Chinese Pharmacopoeia was not less than 0.35.Therefore,the chlorocalcin could be a potential source of cantharidin.3.UPLC-QTOF-MS Qualitative Analysis of High Resolution Mass SpectrometryA total of 144 components were obtained by UPLC-QTOF-MS high-resolution mass spectrometry for the qualitative analysis of Cyanine.All data used the Mass Lynx 4.1 software(Waters Corporation,Milford,MA,USA).This software analysed,processed and was shown to contain many isomeric substances.After comparing the UNIFI library and consulting the references,49 components were identified and their names were obtained.Among them,the active substance in the green genisteinwas seen to be abundant.The effect and influence of these substances need to be further studied in the future.Due to the lack of research reports on the analysis of Cantharides,there is no more in-depth mapping to compare and analyze the components of Cantharides.A large number of substances in Meloidea virga L.were still not identified by MS.In the future,many methods should be combined in order to research and explore these compounds.4.Antitumor and Antibacterial Activity Analysis of Cantharidin and Some of its DerivativesIn Guangxi there is a high incidence of nasopharyngeal carcinoma.In this paper,highly differentiated nasopharyngeal carcinoma(CNE1),poorly differentiated nasopharyngeal carcinoma(CNE2)cells,cervical cancer cells(HELA)and human prostate cancer cell line(DU145)were selected for anti-tumor and antibacterial activity analysis.A normal cell line(293T)of human embryo renal cell line was selected for comparison.By using MTT and CCK8 assays,four types of tumor cell lines,were selected for the inhibition test in vitrowith the following compoundsmethylcantharidine,cantharidin,norcantharidin and sodium norcantharidin,.The results showed there were marked inhibitory effects of cantharidin,norcantharidin and sodiumnorcantharidin on tumor cells.The IC50 was less than 17,but they were also harmful to human normal cells and methylcantharidin was the most significant of the 4 compounds.Cantharidin,norcantharidin and sodiumnorcantharidin were less harmful to human normal cells than the four tumor cells.Norcantharidin and its derivatives have been rarely reported in the field of bacteriostasis.In view of some drug-resistant bacteria in clinical settings,some antibacterial studies were carried out in this paper.Cantharidin,norcantharidin,sodium norcantharidin and methylcantharidamine were selected for these the experiments.The antimicrobial activities of six common clinical drug-resistant bacteria was tested.These were against Proteus spp.,Salmonella enterica,Pseudomonas aeruginosa,Bacillus subtilis,Staphylococcus aureus and Escherichia coli.The results were all negative.