Design,Synthesis And Antitumor Application Of Coumarin Furopyrimidinone Hybrid Derivatives

Background:Tumors pose a huge threat to human health and survival and greatly reduce people’s quality of life.It is one of the most important social problems facing the world.Currently,the main treatment for cancer is chemotherapy.Although a variety of chemotherapeutic drugs have been used clinically and have achieved certain results,there are still many problems in anti-tumor therapy,such as the unsatisfactory therapeutic effects of drugs on solid tumors,large toxic side effects,and tumor resistance.Therefore,it is of great significance to find new anti-tumor drugs with low toxicity,high efficiency,high selectivity and safety.Coumarin is an important oxygen-containing heterocyclic compound with a pyrone structure.It is widely distributed in nature and is a natural active compound with high bioavailability and low toxicity.In the early stages,our research team effectively synthesized a series of novel furanopyrimidine derivatives under mild conditions.Through the inhibition test on the proliferation of several lung cancer cells,it is shown that the target compound has potential anti-tumor activity and has further research value.Objective:In this study,based on the principle of drug design and splicing,with natural active substances coumarin and furanopyrimidinone as the pharmacodynamic framework,new types of coumarin furanopyrimidinone hybrid derivatives were designed and synthesized,and they were screened for anti-tumor activity in vitro to obtain Lead compound with potential anti-tumor activity.Methods:Based on the preliminary research foundation of the research group,under mild conditions,the polysubstituted furan pyrimidinone compounds are synthesized efficiently with readily available raw materials,and the main intermediate furan pyrimidinone hydrazide compounds 6a-6n are further synthesized.Coumarin-3-carboxylic acid ethyl ester was synthesized by Knoevenagel condensation method,and then the 3-formyl chloride coumarin intermediate 10a-10b was obtained by hydrolysis reaction and reaction with thionyl chloride.Finally,using the principle of drug combination,the furanopyrimidinone hydrazide and the coumarin formyl chloride are connected to prepare the target compound.The CCK-8 method was used to evaluate the anti-proliferative activity of the compounds on different tumor cell hepatoma cell lines（HepG2）and cervical cancer cell lines（Hela）,and the apoptosis effect was detected by flow cytometry.Results:In this study,a total of 34 new compounds were synthesized,including 14furopyrimidinone hydrazide derivatives and 20 coumarin furanopyrimidinone hybrid derivatives.Their molecular structures were all characterized by ¹H NMR,¹³C NMR and ESI-MS.In vitro anti-tumor activity tests show that the coumarin furanopyrimidinone hybrid derivatives 11a-11f and 12a-12n have moderate to significant cytotoxic activity on HepG2 and Hela cells.Among them,compound 12i showed good inhibitory activity on HepG2 and Hela cells,with IC₅₀of 4.87μM and8.75μM,respectively.Compound 12n has the strongest inhibitory activity on Hela cells,with an IC₅₀ value of 7.66μM.Compound 11b also showed good anti-tumor activity against HepG2,with IC₅₀of 7.72μM.In order to analyze the correlation between the anti-tumor activity of the target compound and the two-part pharmacodynamic framework of coumarin and furanopyrimidine,we also screened the anti-tumor activity of intermediates 6a-6n,and the results showed that 6a-6n also has certain anti-tumor effects.Among them,6i also showed good inhibitory activity on HepG2 and Hela cells,with an IC50 value of 8.53μM and 20.15μM.Through the comparison of the activity results,the activity of the hybrid derivatives has been significantly improved,indicating that the natural fragrance It is feasible to hybridize the pharmacodynamic framework of legumes with the pharmacodynamic framework of furanopyrimidine.Preliminary structure-activity relationship analysis shows that the anti-tumor activity of the target compound is related to the different substituents of furo[2,3-d]pyrimidinone:the different amine activity sequence introduced at the C-2 position is:dipropylamine>n-butylamine>n-propylamine>diethylamine>morpholine.At the same time,the introduction of a phenyl group on the benzene ring at the C-3 position of the pyrimidine ring greatly enhances its anti-tumor activity.In addition,the introduction of alkyl substitution at the C-7 position of coumarin can significantly increase the inhibitory activity of the compound.Flow cytometry analysis further showed that compound 12i exerts its anti-proliferative effect by blocking cells in the G2/M phase of the cell cycle and inducing apoptosis.The results show that compound 12i can induce apoptosis in a concentration-dependent manner.Conclusion:In summary,based on the principle of drug splicing,this study synthesized a series of coumarin furanopyrimidinone hybrid derivatives with a simple and efficient method.The anti-tumor activity screening and structural activity analysis showed that the natural coumarin pharmacodynamic skeleton It is feasible to hybridize with the pharmacodynamic framework of furanopyrimidine.The results of this study provide a basis for the subsequent further structural optimization,and also lay a certain material basis for the structural modification of the natural active substance coumarin.