Design,Synthesis And Activity Evaluation Of 2,3,4,5-tetrahydro-1H-pyrido[4,3-b] Indoles Derivatives

Nausea is a serious threat to the whole human health.However,the incidence and mortality of nausea are still rising sharply.Drug therapy can not achieve good therapeutic effect on solid tumors because of its individual differences,toxic and side effects,more targets and drug resistance.Therefore,it is urgent to find a new target antitumor drug with high efficiency and safety.Carbazoline alkaloid is a kind of natural alkaloid which widely found in plants with antibacterial,anti-tumor and anti thrombotic activities.The small molecule tyrosine kinase inhibitors with the tetrahydro gamma-Carbazoline exhibit well antitumor activity,however,there are some defects such as many targets,large toxic and side effects.Therefore,it is of great significance to find new high efficient and low toxic four tetrahydro gamma-carbazoline derivatives against the development of tumor drugs.Natural gamma carbazoline alkaloids are less distributed in nature,so most of the studies of gamma carbazoline derivatives are synthesized artificially.On the basis of previous studies,we designed principle of sulfonamide derivatives and amine derivatives by using drug parallel principle and used Hinsberg reaction and Hofmann alkylation synthesismethod to obtain the target compounds,evaluated the activities of two kinds of derivatives,according to the results of activity evaluation for molecular docking and dynamics simulation study.The results showed that two derivatives had strong anti-tumor activity.The main results were as follows:We synthesized 8-bromo-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indoles(intermediate 2)based on previous research,through the structure modification of nitrogen atoms on indole ring after Boc protection(intermediate 3),we designed and synthesized 4 sulfonamide derivatives and 3 amines derivatives,inciuding tert-butyl-8-bromo-5-(phe nylsulfonyl)-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4a),tert-but yl-8-bromo-5-tosyl-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4b),t ert-butyl-8-bromo-5-[(4-chlorophenyl)sulfonyl]-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4c),tert-butyl-8-bromo-5-[(4-nitrophenyl)sulfonyl]-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4d),tert-butyl-8-bromo-5-methyl-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4e),tert-butyl-8-b romo-5-benzyl-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylate(4f),tert-b utyl-8-bromo-5-phenethyl-3,4-dihydro-1H-pyrido[4,3-b]indole-2-carboxylat e(4g),the structure of all compounds were characterized by MS,13C-NMR,1H-NMR,and elemental analysis.The synthesis conditions of two kinds of compounds were optimized and the action mechanisms werre studied.The selected synthetic route was simple,the compound yield was relatively high.This experiment selected all designed and synthesized2,3,4,5-tetrahydro-1H-pyrido[4,3-b] indole derivatives and intermediates to stimulate the proliferation of human cervical cancer cell Hela,human non-small cell lung cancer cell A549,human hepatoma cell Hep G2,human breast cancer cell growth of MCF-7.The inhibitory effects of2,3,4,5-tetrahydro-1H-pyrido[4,3-b] indole derivatives and intermediates on the proliferation of the above four kinds of cancer cell lines were detected by MTT method in 24 h and 48 h respectively.The results showed that the activities of all the derivatives were higher than two intermediates,compound 4c had the strongest inhibitory activity to the proliferation of A549 cell lines,the IC50 value of compound 4c was 9.42? M.In 24 h and48h,all target compounds had no time dependence on the growth inhibition of four different tumor cell lines.Flow cytometry analysis showed that compound 4c can promote the apoptosis of A549 cells and inhibite the cell cycle of A549 cells in a concentration dependent manner.In this experiment,molecular docking and molecular dynamics simulation were used to verify the experimental results of the evaluation of the activity in vitro.The results of molecular docking showed that sulfonyl or carbonyl groups of sulfonamide derivatives can form multiple hydrogen bonds with amino acid LYS1110,and the-CDOCKER energy(kcal·mol-1)value of intermediate 2,3 were lower than all new derivatives which corresponded to the results of antitumor activity in vitro.The results of Molecular dynamics simulation showed the combination between thederivatives and the target enzyme 3EFJ tended to be stable in 8ns,and the RMSD value was near 2.5A,which proved the rationality and stability of molecular docking.In summary,there were 9 derivatives and intermediates of2,3,4,5-tetrahydro-1H-pyrido[4,3-b] indole that synthesized in this experiment,mechanism exploration and optimization of the reaction involved in the experiment were studied,the synthesized derivatives had obvious inhibitory effects on the growth of four kinds of tumor cells and compounds 4c with the most antitumor activity could promote the apoptosis of A549 cells and inhibite the cell cycle of A549 cells in a concentration dependent manner,molecular docking and molecular dynamics simulation illustrated,verified the antitumor activity of all derivatives.