Study On The Anti Breast Cancer Activity Lead Compounds Based On The Chiral Hexahydropyridazine Backbone

Aim:Based on the 1,2-dioxane structure isolated from Lonicerae Japonicae Flos,the entirely new chiral hexahydropyridazines is synthesized by secondary amine-catalyzed organocatalytic cascade reaction with the help of scaffold hopping design.An in vitro assay of antitumor activity of the chiral products and its derivatives is conducted,and then clarify its mechanism.Methods:1.We performed a cascade reaction from aldehydes,nitroolefins,and azodicarboxylates to accesse the aza-shuangkangsu derivatives.The structure was identified by ¹H-NMR,¹³C-NMR and HRMS,the absolute configuration was determined by X-ray single crystal diffraction,and the enantiomeric excess was analyzed by HPLC with a chiral stationary phase.Thus a polysubstituted chiral hexahydropyridazines library is established;2.The in vitro MTT assay was used to determine the antiproliferative activity of the tested compounds in MCF-7 breast cancer cells and HCT116 colon cancer cells.Cell proliferation activity and cell morphology were taken as evaluation indexes to carry through the antitumor activity in vitro at cellular level;3.A systematic drug related pivotal protein interaction network was established by means of bioinformatics technology;4.Flow cytometry and fluorescence staining were used to detect the apoptosis and the cell cycle in vitro,Western blot assay was adopted to detect the expression of the related target proteins regulating apoptosis,so that we can do a further research of molecular mechanism of the compounds that have perfect antiproliferative activity for MCF-7 cells.Results:1.Seventeen new compounds were synthesized with high enantioselectivity.?4a-4eH and 5a-5c?The structure of the target compounds all were correct which have been comfirmed by ¹H-NMR,¹³C-NMR,HRMS analysis,and ee determined by chiral HPLC,which were higher than 90%.2.The antitumor activity in vitro showes that all compounds had a certain antiproliferative activity in tumor cell cultivition,which was positively correlated with dose dependence.The effect of 5c is the most remarkable within all compounds.Conclusion:We have developed a versitile organocatalytic cascade reaction involving a Michael-amination-hemiaminalization relay,and used it to assemble a densely functionalized aza-shuangkangsu derivatives.Starting from aldehydes,nitroolefins,and azodicarboxylates,we obtained a library of bioactive products in moderate to good yield with high stereoselectivity.The synthetic chiral hexahydropyridazines have a certain effect on the proliferation of MCF-7 breast cancer cells and HCT116 colon cancer cells by inducing apoptosis.