| Matrine alkaloids have anti-tumor activity,but it is difficult to separate matrine alkaloids because of the complex components in natural medicines.Therefore,it is of great significance to develop an efficient extraction technology for screening matrine alkaloids.The research ideas of this topic are as follows: The active component matrine in Sophora alopecuroides L.was used as template molecule to enrich and separate the active component groups in Sophora alopecuroides L.extract by molecular imprinting-solid phase extraction technology.At the same time,a set of chromatographic method for quantitative analysis of MISPE-HPLC was established by combining the structure analysis of the enriched components with liquid chromatography-mass spectrometry(LC-MS).Then,computer-aided drug design was used to select the most potent matrine inhibitors for Topo I.In order to establish a rapid and simple method for enriching trace active ingredients from the complex system of traditional Chinese medicine,the anti-tumor active ingredients of alkaloids in Sophora alopecuroides were studied by molecular imprinting and docking techniques.Its main work includes the following three aspects: 一.Preparation and characterization of matrine molecularly imprinted polymer: With matrine as template molecule,the molecularly imprinted polymer with good morphology and properties and particle size in the range of 600-900 nm was obtained by precipitation polymerization.The morphology and structure of the imprinted polymer were observed by transmission electron microscopy,scanning electron microscopy and infrared spectroscopy,and its adsorption and molecular recognition properties were characterized by dynamic/static adsorption experiments and specific adsorption experiments.The experimental results showed that the imprinted polymer had specific recognition and reached adsorption saturation at 200 minutes(Qmax=49.97mg/g).Therefore,it can be used as a solid phase extraction agent to separate and enrich the low content matrine and its analogues in medicinal plants.二.Separation and enrichment of four matrine alkaloids in Sophora alopecuroides by MISPE-HPLC chromatography: In order to rapidly separate the active alkaloids from Sophora alopecuroides,this experiment combined with liquid chromatography-mass spectrometry(LC-MS)to analyze the compounds.The results showed that matrine,oxymatrine,sophocarpine and Oxysophocarpine in Sophora alopecuroides extract were separated by directional method.Molecular imprinting solid phase extraction(MISPE)and high performance liquid chromatography(HPLC)were used to quantitatively analyze the four components.The results showed that the linear relationship of the four alkaloids was good(r < 0.9974),and the average recovery was 89.6%-98.0%.Compared with C18 column,this column can effectively remove the matrix effect and achieve the specific separation,enrichment and detection of matrine compounds in Sophora alopecuroides extract.三.Molecular docking of matrine alkaloids with Topo I: In this study,the binding sites and interactions between ligands(matrine)and receptors(DNA topoisomerase I)were analyzed by molecular docking method in computer-aided drug design(discovery studio 2.5).The results showed that the binding ability of Oxysophocarpine and sophoridine to target protein was higher when the ligand compounds were put into the AC1 cavity of the receptor protein active site,and the active amino acid A: LYS425 and other residues play the main role.On the other hand,protein loci 3 and 6 were found by the software,and their positions were stabilized by interaction with ligand small molecule Oxymatrine and sophoridine through hydrocarbon interaction.However,whether loci 3 and 6 are active loci needs to be confirmed by subsequent pharmacological experiments.Using computer-aided drug design as the pre-screening prediction not only greatly shortens the time and saves the cost,but also has important research significance for accelerating the development of active ingredients of traditional Chinese medicine. |
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