| ObjectiveBased on the analysis of the synthetic route described in the literature,we designed a synthetic route suitable for industrialized production.To investigate the effect of sorafenib on the proliferation,apoptosis of MHCC97-H human hepatocellular carcinoma cells in vitro and its mechanism by reducing expression of PRL-3 protein and cell invasion and metastasis.MethodsUsing 4-(4-aminophenoxy)-N-methylpicolinamide,4-chloro-3-(trifluoromethyl)aniline and triphosgene as starting materials,we got the final targeted product by one-pot reaction.The reaction conditions in the synthetic proc ess were examined in detail.Different densities of sorafenib were applied to MHCC97-H human hepatocellular carcinoma cells,and the MTT assay was used to observe the inhibiting effects of sorafenib on the cell proliferation at 24 h,48 h,72 h to screen the optimum concentration of sorafenib and intervention time.Giemsa was used to observe the morphological changes of cells stained,Western-Blot was used to detect the protein expression of PRL-3,Transwell assay was used to detect the invasion of tumor cells.ResultsThe effects of reaction conditions,such as reaction solvent,reaction temperature,material ratio and feeding sequence,on the yield of Sorafenib were investigated in this experiment.Finally,Sorafenib was prepared with the optimum conditions.Compared with the literature(93%),the yield of sorafenib was increased to 94.2%,and the purity of the product was 98.5% by HPLC.The structure was verified by1 H NMR,13 C NMR and MS.The optimum sorafenib experimental concentration and intervention time were 16 μmol/L and 48 h,respectively.Sorafenib can induce apoptosis of MHCC97-H hepatocellular carcinoma cell line,hepatocellular carcinoma cell proliferation inhibition rate correlated positively with time and dose,PRL-3protein also appeared decreased at different extent.After the expression of PRL-3 decreased,hepatocellularreaction conditionscarcinoma cell invasion ability was significantly inhibited.Conclusions(1)The one-pot method is effective,mild and highly productive,and there was no need to use column chromatography.The process may be suitable for industrial applications.(2)Sorafenib can reduce the invasion of tumor cells and induce apoptosis of hepatoma cells.(3)Sorafenib could decrease the expressionreaction conditions of PRL-3protein in MHCC97-H hepatoma cell line.(4)Sorafenib may decrease the invasionreaction conditions of cancer cells by down-regulating the PRL-3 protein. |
