Screening For Anti-HBV Drugs And Research On It's Anti-HBV Activity

Objective: To screen natural products and other compounds for discovering anti-HBV lead compounds by using HepG2. 2. 15 cell culture system in vitro. The initial research on the relationship between the structure and the effect of some active compounds and the anti-HBV mechanism of the active compounds were done. A new quantitative assay developed to detect HBV covalently closed circular (HBV cccDNA) by SYBR green dye and real-time polymerase chain reactions was described.Methods: Cytoxicity of compounds on HepG2. 2. 15 cell was detected using MTT, the levels of HBsAg or HBeAg in the culture supernant of 2.2. 15 cells were tested by ELISA. Adefovir was selected as a positive anti-HBV compound. Total 358 compounds were screened in the primary screening. Among the active conpounds found in the primary screening , resveratrol and flavonoids were selected for further research on anti- HBV mechanism and it's initial structures-activity relationship. 15 resveratrol derivatives and 16 flavonoids derivatives were collected for the research. The numbers of total HBV DNA copy were quantified by Taqman probe real-time PCR. FACS was employed to test the effect of resveratrol on the apoptosis of HepG2. 2. 15 cell.The main process of establishing method to quantity the numbers of HBV cccDNA copy was as follow: constructed HBV cccDNA plasmid as a standard positive sample , then the numbers of HBV cccDNA copy were quantified by SYBR green I dye real-time PCR.Results: (1) 820 times screening were completed. Therapeutic Indexs (TIs) of 154 compounds were greater than 1.