Design, Synthesis And Evaluation Of Anticancer Selectivity Of Ligustrazine-bile Acid Derivatives

Backgroud:Liver cancer is one of the most common causes of cancer death in the world.Unfortunately,there is no efficient treatment of liver cancer.Currently,due to the poor distribution and stability,serious side effects on other organs,chemotherapy for liver cancer has been greatly hampered,therefore,large efforts have been made for newly anti-cancer drugs in the past decades.According to Traditional Chinese Medicine(TCM)theory,the liver cancer are mostly "Archives of Jia accumulation",the intrinsic toxic heat,blood stasis is the major cause of liver cancer.Therefore,Traditional Chinese detoxify drugs:bezoar and bear bile,endogenous hepatocyte-specific ingredients,with the function of detoxification and promoting blood circulation,were used in the treatment of liver cancer.In this study,Ligustrazine(TMP),commonly used in clinical as adjuvant drug to cancer treatment,was selected as parent nucleus,and bile acids(BAs)were used as drug carriers,a series of TMPBA derivatives were synthesized.Wherein the Ligustrazine-Hyodeoxycholic acid derivative PH5-2(TMP-HDCA)can significantly inhibit a variety of tumor cells proliferation,and display less toxic to normal cells.Research methods:To demonstrate the organ specificity effect of bile acids as drug carriers,the cytotoxicity of 6 bile acids were investigated against both tumor and normal cells.Then,a series of newly TMP-BA derivatives were designed,synthesized.Combined with MTT assays,the derivatives’ selective cytotoxicity,dose-response effect and structureactivity relationships were evaluated using many cell models,including hepatoma cell line HepG2,human colon carcinoma cell line HT-29,gastric cancer cell line BGC823,cervical cancer cell line Hela,hepatocyte line L02 and canine kidney cell line MDCK.And the premechanism of leading compounds on HepG2 cells was studied by Giemsa staining,DAPI staining,AO/EB staining,apoptosis analysis and mitochondrial membrane potential assay.Results:Most of BAs displayed high selectivity related to different cell lines and drug concentrations.Then,13 newly TMP-BA derivatives were synthesized,their chemical structures were confirmed by Hl-NMR,13C-NMR and high resolution mass(HR-MS).And the MTT results showed that most of TMP-BA derivatives displayed stronger toxicity against broader spectrum of tumors,especially HepG2 and HT-29 cells,while friendly to MDCK cells,Among 13 TMP-BA derivatives,PH5-2 was identified to be the most significant candidate targeting enterohepatic cells(HepG2:IC50=6.805μM,HT29:IC50=7.336 μM).The preliminary mechanistic studies indicated that the PH5-2 might inhibit HepG2 cell proliferation,induce loss of the mitochondrial membrance potential,cause cell damage and result in cells apoptosis.Conclusions:13 related derivatives were designed and synthesized by combining TMP and BAs via amide bonds.Then we systematically evaluated the antitumor activities of 6 bile acid and the above derivatives,to prove that BAs were capable to deliver drugs as potential hepatic carcinoma drug carrier.And PH5-2 was the first screened out to be a hepatic carcinoma targeting leading compound.This paper further validate the TCM theory,the application of modern drug design combination principle and the active ingredient in TCM to optimize the structure,and then found that efficiency and low toxicity of anticancer drugs is possible and meaningful.