Synthesis And Anti-inflammatory Evaluation Of Spirooxindole Compounds Containing Benzimidazoisoquinoline

Spirooxindole is the core skeleton of many natural alkaloids.It can be combined with other heterocyclic compounds to obtain more spirooxindole derivatives with excellent biological activity for new drug development.Benzimidazole and isoquinoline are also two privileged structure units,which widely exist in many natural products and drug molecules with a variety of biological activities.In order to study the pharmacological activities of the new skeleton obtained from the combination of these privileged structure units,spirooxindole derivatives containing benzimidazoisoquinoline were synthesized in this dissertation,and their pharmacological activities were investigated.The dissertation is divided into three chapters:Chapter 1:The progress in the synthesis and bioactivity of some benzimidazo-isoquinoline and spirooxindole derivatives are reviewed.Chapter 2:A series of spirooxindole derivatives containing benzimidazo-isoquinoline were synthesized starting from the N-alkylation of isatin,then nucleophilic addition with the benzimidazole-derived carbanion and finally intramolecular Friedel-Crafts cyclization reaction.And the strategy was extended to the synthesis of structurally diverse compounds.29 target compounds were synthesized and their structures were characterized and confirmed by ¹H NMR,¹³C NMR,and HRMS.Chapter 3:After a preliminary screening of the biological activity,many target compounds with potential anti-inflammatory activity were determined and further evaluated via LPS-induced RAW 264.7 cells model.Firstly,compounds exhibiting low toxicity against RAW 264.7 cells were screened by the MTT method.Then,the ability of these compounds to inhibit the release amount of NO from LPS-induced RAW 264.7 cells was tested by the Griess assay.Based on the survival rate of cells which were treated with compound and LPS,compounds 4e and 4i with low toxicity and significant inhibitory effect on the release amount of NO from LPS-induced RAW264.7 cells were selected for further investigation on their anti-inflammatory mechanism.The inhibitory effects of 4e and 4i at different concentrations（5,10,20μM）on the release amount of NO and inflammatory factors from LPS-induced RAW264.7 cells were further investigated by Griess assay and RT-q PCR.The results showed that 4e and 4i could inhibit the release amount of NO and the m RNA expression level of IL-6,COX-2,i NOS,IL-1β.The results of Western blotting suggested that 4i could down regulate the expression of i NOS and TLR-4 protein in LPS-induced RAW 264.7 cells,inhibiting the phosphorylation level of JNK in the MAPK signaling pathway.The results indicated that 4i can exert its anti-inflammatory effect by regulating the MAPK signaling pathway.