Synthetic and analytical method establishment of 3-(4-chlorophenyl)-7-[JP]\[2-(piperazin-l-yl)ethoxy\]-4H-chromen-4-one

Abstract

In order to solve the problem of insufficient antitumor activity of natural daidzein (DAI),the 4 'and 7 positions of DAI were modified.Firstly,3-(4-chlorophenyl)-7-hydroxy-4H-chromen-4-one was prepared from resorcinol and p-chlorophenylacetic acid by friedel crafts acylation,carbonization and ring closing reaction.The 3-(4-chlorophenyl)-7-\[2-(piperazin-l-yl)ethoxy\]-4H-chrome-4-one (CPEO-43) was synthesized by substitution reaction of 3-(4-Chlorophenyl)-7-hydroxy-4H-chromen-4-one with dibromoethane and piperazine.Column chromatography was used to separate the target product from impurities.Secondly,FT-IR,1</sup>H-NMR,13</sup>C-NMR and MS were used to identify the compounds.Thirdly,MTT assay was used to detect the inhibitory effect of compound CPEO-43 on A549 lung cancer cells and HCT116 colon cancer cells.Finally,an ultra high performance liquid chromatography (UPLC) method was established for the analysis of CPEO-43 plasma samples.The results show that the structure of the synthesized compound is consistent with that of the target compound confirmed by FT-IR,MS,1</sup>H-NMR and 13</sup>C-NMR.The inhibitory rates of DAI (10 μmol/L) on A549 lung cancer and HCT116 colon cancer cells are 4.421% and 5.601%,respectively.The inhibitory rates of the same concentration of CPEO-43 on A549 lung cancer cells and HCT116 colon cancer cells are 58.43% and 72.03%,respectively.The IC50 values of CPEO-43 are 2.51 μmol/L and 0.87 μmol/L,respectively.The established UPLC method has no interference from endogenous substances.The linear relationship is good in the range of 0.5~10 μg/mL.The intra-day precision and inter-day precision are less than 10%.The average recoveries of low,medium and high plasma samples of compound CPEO-43 are in the range of 92.98%~100.1%.The antitumor activity of compound CPEO-43 is significantly higher than that of DAI.The established UPLC assay method can easily and rapidly determine the content of CPEO-43 in plasma,and the results are accurate and reliable,provide a theoretical basis for improving the antitumor activity of DAI,and provide method basis for drug clinical testing.

Keywords

• other disciplines of pharmacy; daidzein; soybean isoflavone derivatives; antitumor activity; ultra high performance liquid chromatography; nuclear magnetic resonance; mass spectrum