A Preliminary Study On Anti-hepatoma Mechanism Of Tipranavir

BackgroundHepatocellular carcinoma is one of the most common and highly malignant malignancies,Ranking the fifth highest of global malignancy,the incidence of malignant tumors ranked second in China,a serious threat to human health and life.Radiofrequency ablation,chemotherapy,surgical resection,liver transplantation and other commonly used methods of treatment of liver cancer,gradually form a the comprehensive treatment model based on surgical resection.However,although the current treatment can be effective in the treatment of local lesions,but patients easy to postoperative recurrence.radiotherapy,chemotherapy is easy to produce resistance.Molecular targeted anti-tumor drugs have a certain degree of superiority in reducing the traditional side effects of chemotherapy drugs and improve the efficacy,this drugs have a specific characteristics of tissue distribution,which can reduce the average distribution of drugs in the body,reduce the dosage and the number of drugs.Thereby,increasing the drug treatment index,reducing its systemic serious side effects.Silencing information regulation factor 1(SIRT1)is a class of deacetylase that relies on nicotinamide adenine dinucleotide(NAD+).Involved in cell energy metabolism,stress tolerance,differentiation,aging and apoptosis and other physiological activities.According to the literature,the expression level of SIRT1 in hepatocarcinoma tissue was significantly increased,and the small molecule compound Tipranavir was screened by virtual screening.It was found that Tipranavir had a strong inhibitory effect on hepatocarcinoma cells by vitro and vivo experiments of anti-hepatocarcinoma activity.According to the Pharmacopoeia and the literature,Tipranavir is an inhibitor of CYP3A4 liver enzyme,which is the most of the liver enzyme.This provides a thought that,whether Tipranavir is acting on SIRT1 or acting on CYP3A4,or whether the synergies lead to inhibition of proliferation of hepatocellular carcinoma cells,or that there may be other targets of action,which is worthy of to discuss the problem.Research objectiveBased on the Sybyl2.1 platform's Surflex-Dock module and the Ligand Fit docking module of Discovery Studio3.0 platform,we compared the results of Tipranavir with SIRT1 and CYP3A4 through docking studies,then construct HepG2 SIRT1 knockout Cellsand HepG2 CYP3A4 knocked out cells,monoclonal screening HepG2 knockout cell strains.Then through CCK-8 proliferation inhibition experiments to verify inhibition rate of HepG2 cells and knockout cells,which laid a foundation for the study pathway of Tipranavir inhibition of HCC.Research methods1.The Tipranavir was docked with the SIRT1 and CYP3A4 crystal structures based on the CADD platform.2.Construction of HepG2 cells SIRT1 and CYP3A4 knocked out cell lines and verified.3.Screening monoclonal cell of knockout cell lines and validation.4.The proliferation inhibition rate of HepG2 and its knockout cells was detected by CCK-8 proliferation inhibition method.5.Reverse molecular docking based on the Id Target and the PharmMapper platform using Tipranavir as a ligand.Research resultsIn this study,the inhibition rate of HepG2 and its knockout cells was detected by CCK-8 proliferation inhibition assay.It was concluded that Tipranavir had a certain inhibitory effect on SIRT1 and CYP3A4 knockout cells,but it was not obvious.Tipranavir does not have much cytotoxicity to normal liver cells.