Study On The Histone Deacetylase Inhibitors Of Polyamines:Screening And Anti-Lung Cancer Effect

Lung cancer is one of the most common malignant tumors.According to statistics,the incidence of lung cancer in the world was 12.7%,the mortality rate was about 18.1%.The incidence of lung cancer in China was 24.7%,with a mortality rate of 30%,ranking first in the world.At present,there is still lack of effective drugs in the treatment of lung cancer,especially lung squamous cell carcinoma.Screening for safe and effective treatment of lung cancer has become an urgent public health problem.This study used the histone deacetylase inhibition of molecular docking,which based virtual screening,mass spectrometry and NMR analysis of in vitro activity evaluation based on the technology.With cystamine and spermine as the lead compound,the derivatives,evaluate the effect of anti lung cancer.1.The virtual screening method of histone deacetylation inhibitor was established.By using the method of computer virtual screening,HDAC2,the first class of HDAC,was selected as the target of tumor resistance from the protein crystal database PDB;Through the introduction of benzene,thiophene,furan and pyrrole groups at the ends of cystine and spermine,a total of 51 derivatives with multi amine structure were designed,and selected a group which has been listed on the protein deacetylase inhibitor chidamide(CS055)as a reference;Sybyl-X2.0 software was used to carry out molecular docking experiments of receptor protein and ligand libra ry compounds.The results of comprehensive molecular docking showed that 6 compounds had more inhibitory effect on histone deacetylase than positive control drugs.The synthesis method and the structure elucidation of the candidate compounds were screened out.2.Synthesis and structure elucidation of 6 candidate compoundsThe synthesis of the candidate compounds was carried out by the addition reaction of spermine and cysteamine with aromatic aldehyde,thiophene aldehyde,furan aldehyde and pyrrole aldehyde.The structures of 6 compounds were identified by NMR and mass spectrometry.The final 6 compounds were :N,N’-(disulfide-2,1-diethyl)bis(2-thiophene methylamine)(1b);N,N’-(disulfide-2,1-diethyl)bis(2-fluorobenzoylmethylamine)(2b);N,N’-(disulfide-2,1-diethyl)bis(2-Furanmethylamine)(3b);1,16,-2-(2-pyrrole)-2,6,11,15-Tetrazahe xadecane(4b);1,16,-2-(2-Furan)-2,6,11,15-Tetrazahexadecane(5b);1,16,-2-(4-hydroxylphen yl)-2,6,11,15-Tetrazahexadecane(6b).3.Found the 4 compound that had inhibitory effect on lung squamous cell carcinoma.Using the method of CCK-8,chidamide as the positive control drug,the candidate compounds were synthesized by in vitro inhibition of lung squamous cell carcinoma NCI-H520 and normal human bronchial epithelial cells 16 HBE,The appropriate cell density of NCI-H520 cells and 16 BHE cells were selected and the culture time after entering CCK-8,the 24 h restrained effects of candidate compounds on the above tumor cell lines(in 7 concentrations:0.1、1.0、10、25、50、100、120μMoL/L)was investigated with CCK-8 assay,and its median inhibitory concentration(IC50)was calculated.The experiment fo u nd t ha t t he d e ns it y o f N C I-H 5 20 c e lls a t 9 6-w e ll p la t e s t ha t w a s 2500/holes(1~3day),Add CCK-8 to culture for 1 h,the O D value in the range of 0.85~1.13.And the density of 16 HBE cells at 96-well plates that was 5000/holes(1~3day),Add CCK-8 to culture for 1h,the O D value in the range of 1.05~1.59.Select 9 compounds and positive drugs:number1、2、3、4、5、6、10、12、14、19.The IC50 values of NCI-H520 were 222、152、369、5、12、9、2822、157、143、45μMoL/L,The IC50 values of 16 HBE were 201、96、80、71、2076、30、2923、143、711、39μMoL/L.J1-137、J2-138、G1-132 and N,N’-(disulfide-2,1-diethyl)bis(2-thiophene methylamine)(1b)were screened out.These compounds have a strong inhibitory effect on human lung squamous carcinoma cell line NCI-H520 in vitro.The inhibitory effect of J1-137 compound on human lung squamous carcinoma cell line NCI-H520 was obvious.The IC50 values are 5μMoL/L.Thus the inhibition effect of 16 HBE on bronchial epithelial cells was not significant.The IC50 values are 2076μMo L/L.It provides a reference value for the study of polyamines.