| This paper was composed by two parts.The first part was about the study on the quality analysis of Epimedium.The second part was about the chemical synthesis of quinolone derivatives and the antitumor effect research.(Part 1)Epimedium was a commonly used traditional Chinese medicine,contained the version of the calendar year "pharmacopoeia of the People's Republic of China".The traditional Chinese medicine was widely used in the clinical compatibility of traditional Chinese medicine for many diseases,and also for many Chinese patent medicine and health medicine raw materials.In order to control the quality of epimedium effectively,the content analysis of the effective components group was conducted in order to provide experimental basis for the revision of epimedium quality standard in Chinese pharmacopoeia.The content of effective components group of epimedium was analyzed and studied by modern ultra performance liquid chromatography(UPLC).Five effective constituents of flavonoids from epimedium Herba Epimedii were identified.The established method has good linear relationship,precision,repeatability,stability and recovery are in compliance with the requirements(RSD < 3%).The method has the advantages of short analysis time,good separation effect,less solvent consumption and less solvent consumption.Using the method,the comparative analysis of 14 batches of epimedium effective components group,the results can effectively evaluate the quality of different habitats.Therefore,a reliable quality analysis method for the medicinal materials,can more comprehensively reflect the internal quality.(Part 2)Quinolone was an important active ingredient in many traditional Chinese medicine.According to the literature,the anti-tumor active ingredients of many Rutaceae plants are quinolone compounds.Therefore,the chemical synthesis of quinolone as the lead compound was studied in this paper,in order to find the high efficient anti-tumor compounds,to provide the target compound for the study of antitumor drugs.In this paper,2-quinolones were synthesized by methylation and oxidation under basic condition from quinolines.Nitro-2-quinolones were obtained by nitration.Finally,some 1-methyl-4-cyanide-2-quinolones were synthesized by cine-substitution reaction of some nitro-2-quinolones with potassuim cyanide.The structures were confirmed by 1H NMR,13 C NMR and HR-MS.The antitumor activities of the compounds were investigated by MTT assay on MCF-7,H1299,A549,PC-12,CT-26 and HepG-2.The results showed that some of compounds exhibited better inhibition activities than quinolines.1,8-dimethyl-3,5,7-trinitro-2-quinolone(15)exhibited good inhibition activities against the six cancer cell lines,especially A549 cell with IC50 of 2.05 ?mol·L-1.1-methyl-6,8-dinitro-2-oxo-1,2-dihydroquinoline-4-carbonitrile showed better inhibition activities against A549 and CT-26 with IC50 of 9.34 ?mol·L-1and 18.43 ?mol·L-1,respectively.Take A549 cells as subject,we studied anti-tumor mechanism of 1,8-dimethyl-3,5,7-trinitro-2-quinolone(15)through using Annexin V-FITC/PI double staining and western blot analysis.The results show that compound 15 could significantly inhibit the proliferation of A549 cells,show a dose and time-dependent relationship,and induce apoptosis.The flow cytometry detected that compound 15 could induce the A549 cell apoptosis in a dose range.The expression of Cleaved-Caspase-3 and Bax were up-regulated with the increase of drug concentration,and the expression of Bcl-2 was down-regulated with the increase of drug concentration,by Western blot analysis of compound in A549 cells at different concentrations(0,1.25,2.5 and 5 ?mol·L-1).Therefore,compound 15 might induce A549 lung cancer cells apoptosis through mitochondrial pathway.At the same time,PI3K/AKT/FOXO1 signaling proteins were detected,the expression of p-AKT and p-FOXO1 can be down-regulated,and the expression of Bim can be up-regulated.Therefore,compound 15 might induce apoptosis of A549 lung cancer cells by inhibiting PI3K/AKT/FOXO1 signaling pathway. |
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