Study On The Anti-cancer Activity Of Synthetic Targeted NTCP Inhibitor And Pyrrolizidine Nervosine Ⅶ

Tumor is a major disease that seriously threatens human health and life.The mortality rate of malignant tumors is extremely high.It is still a human dream to find anti-cancer methods and drugs to completely cure cancer.Clinically,a variety of small molecule drugs have shown good anti-cancer efficacy,which have certain advantages in improving the quality of life of patients and extending the survival period.Small molecule compounds as a source,the search for anti-cancer lead compounds with clear mechanism has broad prospects and far-reaching social significance.This research includes two parts as follows:1.Design,synthesis and anti-cancer mechanism of targeted NTCP inhibitorsNTCP（Sodium taurocholate cotransport polypeptide）,as the main receptor for hepatitis virus,has been a star target for the development of anti-hepatocarcinoma drugs.In the research,the lead compound AO-081/40926746(IC₅₀=13.4μM)with good activity against HepG2 cells was screened out from 10 candidate drugs by virtual screening of NTCP inhibitors combined with in vitro activity screening.Under the guidance of the classical drug design principle,thestructural modification of the lead compound was carried out,and 24derivatives were prepared.Among them,the most active compound 35（N-（2-（2-（2-6-chlorobenzo[d][1,3]dioxol-5-yl）methoxy）benzylidene）hydrazinyl）-2-oxoethyl)-4-methylbenzamide)was obtained by MTT assay.After treatment with compound 35,western blot showed that Bax,cleaved-caspase 3,and cleaved-PARP were up-regulated,and Bcl-2,NTCP were all inhibited.The interaction pattern of the compound 35-NTCP was analyzed by molecular docking and kinetic simulation,and the results showed that the compound 35 and the NTCP protein were stably combined.Compound 35 is a novel NTCP inhibitor with good anti-proliferative activity against HepG2 cells.2.Anti-cancer activity of pyrrolizidine alkaloids in the Liparis L.C.RichAccording to literature review,various pyrrolidine alkaloids have excellent anti-cancer activity,but their hepatotoxicity limits their development.In this study,the anti-cancer activity of pyrrolidine alkaloid was studied,which was isolated from Liparis nervosa.Preliminary anti-tumor screening bioassay in vitro showed that Nervosine Ⅶ had excellent anti-proliferation activity on HCT116 cells(IC₅₀=11.7μM),with a concentration-dependent and time-dependent relationship.Hoechst 33258 and Annexin V-FITC/PI staining revealed that Nervosine Ⅶ induced apoptosis in HCT116 cells.Using western blot method,it was found that Nervosine Ⅶ increased Bax expression,decreased Bcl-2 expression,activated caspase 9,caspase 3 and PARP.The apoptosis is ultimately induced by the mitochondrial pathway.