The Synthesis Of 4,5-Substituted Phenyl-4H-1,2,4-Triazole-3-Amine And Their Inhibitory Effect On Lung Cancer Cells

Background: Aminotriazole compounds have a wide range of biological and pharmacological properties,such as antibacterial,antioxidant,antimicrobial,antiviral,anticancer,etc.Triazolium ring is the basic structure of histidine and histamine in living organisms.It is the main group for the expression of biological activity and biological action of these two substances.As the functional group of the active center of many enzymes,the triazole ring structure,which is associated with many important biochemical reactions,plays a very important role in the activities of life.It has been reported that triazole ring is also an active substrate for the preparation of many drugs.Because of this biological property of triazole ring,its derivatives play an important role in the development of medicine,especially in the field of anti-tumor.Objective and significance: In this study,aminotriazole derivatives(TA)were designed and synthesized by means of computer-aided design and synthesis to screen the active structures that can inhibit the proliferation of tumor cells and reduce the toxic and side effects.To explore the inhibitory effect of aminotriazole derivatives on the proliferation and apoptosis of lung cancer cells,so as to provide reference for the study of anti-lung cancer drugs.Methods: Eleven 4,5-substituted phenyl-4H-1,2,4-triazole-3-amine derivatives were synthesized by condensation reaction,sulfur substitution,hydrazine substitution and cyclohexane reaction.The apoptosis induction and its mechanism were observed by MTT,fluorescence staining,flow cytometry and western blot.Results: Through a series of chemical reactions,11 target compounds were successfully obtained,and 11 4,5-substituted phenyl-4H-1,2,4-triazole-3-amine derivatives had different inhibitory activities on different tumor cells,especially for lung cancer.The experimental results showed that the inhibition effect of most compounds was stronger than that of control drugs,especially compound 4c(BCTA)(IC50,A549:1.09 ?M;NCI-H460:2.01 ?M;NCI-H23:3.28 ?M)showed significant inhibitory effect on all three types of lung cancer cells,and was less toxic to normal cells.Under the fluorescence microscope observation,the cell morphology changes,the cell membrane blurs,the cell adhesion.The result of flow cytometry showed that the apoptosis rate of A549 cells induced by BCTA was 55.50%,which was significantly better than that of thecontrol group(5-FU,33.03%).Western blotting showed that BCTA up-regulated the expression levels of pro-apoptotic proteins Bax,Caspase-3 and PARP,while down-regulated the expression levels of anti-apoptotic proteins Bcl-2.Conclusion: 1.Lawesson's reagent and toluene reagent can promote the formation of thiotropic reaction.2.The cycloreaction of aminoformamidine compounds with hydrogen bromide can be treated at low temperature to reduce the metamorphism of aminoformamidine(i.e.self-polymerization)and to increase the yield.3.4,5-substituted phenylaminotriazole derivatives can effectively induce tumor cell apoptosis,among which BCTA has the best inhibitory activity on lung cancer cells,significantly better than5-FU inhibitory activity.4.Western blot showed that BCTA could up-regulate the expression of pro-apoptotic proteins Bax,Caspase-3 and PARP,and down-regulate the expression of apoptotic protein Bcl-2.