Design,synthesis And Antitumor Activity Of α-Aminophosphonate Derivatives Containing Matrine

Objective:So far,cancer treatment is still a major problem,drug resistance,toxicity and other problems still cannot be effectively solved.The aim of this study was to introduce the targeted prodrugα-aminophosphonate group into the active skeleton of matrine,and then screen out efficient and low-toxicity new matrine derivatives with more specific targets and mechanisms of action.Their structures were further optimized through molecular simulation,and explore their antitumor activities and mechanism of action in vitro and in vivo.Method:Using matrine as raw material,α-aminophosphonate group was introduced into matrine skeleton by ring-opening,esterification,hydrolysis,acylation and other reactions,and 20 novelα-aminophosphonate derivatives containing matrine were synthesized.The structure of the compounds was confirmed by ¹HNMR,¹³CNMR,HRMS,and IR.The pharmacokinetics,toxicity and drug-like properties of the compounds were predicted by molecular simulation.The in vitro antiproliferative activity of the target compound against MGC-803（human gastric cancer cells）,Hela（human cervical cancer cells）and Hep G2（human liver cancer cells）and the in vitro cytotoxicity against LO2（human normal liver cells）were evaluated by MTT assay.The in vivo antitumor activity of compound 7k was evaluated by nude mouse transplantation.Finally,the mechanism of compound 7k was investigated by Western blot,Hoechst33258staining,reactive oxygen species（ROS）and mitochondrial membrane potential detection Result:The results of MTT method showed that the synthesized compound had good selectivity to Hela cells with high expression of Mcl-1protein.The results of molecular simulation showed that the binding energy of the target compound with Mcl-1 was high,and the binding energy of 7k and 8k with Mcl-1 was higher than 6k,with 7k being the most significant.The MTT results of compound 7k and 8k showed that compound 7k had better antitumor activity.In vivo xenografts of nude mice,compound 7k had less toxic side effects than cisplatin,and the low-dose and high-dose groups of 7k showed 17%（5mg/kg）and 21%（10mg/kg）tumor inhibitory effects compared with the control group.The results of Hoechst 33258 staining showed that 7k induced apoptosis and morphological changes of Hela cells in a concentration-dependent manner.Reactive oxygen species level and mitochondrial membrane potential detection results showed that compound 7k could induce mitochondrial membrane dysfunction,resulting in high intracellular reactive oxygen species level and oxidative stress,and finally lead to mitochondrial membrane permeability opening and release of apoptosis factors to induce cell apoptosis.Conclusion:A series ofα-aminophosphonate derivatives containing matrine were synthesized.In vitro bioactivity studies showed that the compounds had obvious selectivity against Hela（human cervical cancer cells）,and some of the compounds had significant antitumor activity.In addition,the selected compound had strong binding ability to the Mcl-1 protein.It can also increase the level of reactive oxygen species and down-regulate mitochondrial membrane potential.It has certain antitumor activity and no obvious toxic side effects.The above research results can provide theoretical guidance for the design of novel Mcl-1 inhibitors.