Study On The Virtual Screening Of The Histone Deacetylase Inhibitor And Its Inhibitory Effect On Human Hepatocarcinoma Cells HepG2 In Vitro

Liver cancer,is one of the common digestive cancer,the mortality of primary liver cancer ranked second in China,and ranked third in the world.It has become the third most common malignant tumors just behind gastric cancer and esophagus cancer.It is a serious threat to human health and life,and there is still a lack of effective clinical drugs.Histone deacetylase inhibitors can interfere with the histone acetyltransferase function,it can inhibit tumor cell proliferation and induce cell differentiation and(or)the function of apoptosis.Histone deacetylase inhibitors have become a new hotspot arch of the anti tumor targeting therapy.In this paper,we reseaech the Inhibitory Effect of a series of natural compounds and their derivatives on the histone deacetylase,optimizing the preparation methods of histone deacetylase inhibitors compounds,and the Inhibitory Effect on Human Hepatocarcinoma Cells Hep G2 in vitro.We are summarized as follows:1.The virtual screening method of histone deacetylation inhibitor was established.Select two receptor model of HDAC2 and HDAC8 from the PDB database,they are Histone deacetylase inhibitors and related to the prevention and treatment of tumor.Collection from the literature,or derivatives of the parent nuclear design,we get 369 compound structure as ligands,including 345 lignan compounds,28 polyamine compounds,3 histone deacetylase inhibitors which have been listed on.Using Sybyl-X2.0 software,do the experiment of two receptor moleculars docking with ligands.By consider of docking result and the binding model,finally confirmed six polyamines derivatives as the histone deacetylation inhibitor active compounds,were synthetic preparation next step.2.Select the synthesis method of Histone deacetylase inhibitors compouds.Through literature,obtained the route of Schiff base reaction,and then reduction.Continue to test drug proportion,gas protection,reflux solvent,reaction temperature and reaction time and so on various conditions,select the method of cystamine hydrochloride and spermine as raw material,through the first step Schiff base reaction were reacted with the hydroxyl benzaldehyde,furan formaldehyde,pyrrole aldehyde,and generate imine,then reduction with sodium borohydride in the second step,finally separation and purification.By this method,Prepared 6 compoudswhich parent nucleus is cysteamine and spermine.3.Confirm the structure of the compounds.Using infrared spectroscopy,nuclear magnetic resonance(NMR)and mass spectrum spectroscopy method,as well as to the synthesis of 6 compounds pop,corroborate structure respectively are4,4’-(((disulfanediylbis(ethane-2,1-diyl))bis(azanediyl))bis(methylene))diphenol,2,2’-disulfanediylbis(N-(furan-2-ylmethyl)ethanamine),2,2’-disulfanediylbis(N-((1H-pyrrol-2-yl)methyl)ethanamine),4,4’-(2,6,11,15-tetraazahexadecane-1,16-diyl)diphenol,N1,N1’-(butane-1,4-diyl)bis(N3-(furan-2-ylmethyl)propane-1,3-diamine),N1,N1’-(butane-1,4-diyl)bis(N3-((1H-pyrrol-2-yl)methyl)propane-1,3-diamine).4.The Inhibitory Effect of histone acetylation inhibitors compounds on human hepatocarcinoma cell Hep G2 was tested in vitro.Using MTT assay and 5-FU as positive control,through the test of The Inhibitory Effect of 6 compounds on human hepatocarcinoma cell Hep G2 in vitro,screening out the best inhibition of human hepatocarcinoma cell Hep G2 is2,2’-disulfanediylbis(N-((1H-pyrrol-2-yl)methyl)ethanamine).and from the inhibition rate and IC50 data can determine its activity is slightly better than 5-fluorouracil.