Design And Synthesis Of Lamivudine-polyphenol Prodrugs And Evaluation Of Its Anti-HBV Activity

Hepatitis B virus(HBV)infection is one of the public health issues,which is a worldwide concern,and the most common cause of liver cancer and cirrhosis with high morbidity and mortality.Lamivudine is the first-generation nucleoside analog drug approved by FDA for treatment of HBV,and has a high inhibition rate for HBV-DNA replication.However,the effect of drug resistance is observed with patients in long-term treatment,which limited the therapeutic effect.It is reported that polyphenols have anti-HBV and liver protection effects.Protocatechuic acid is a polyphenolic compound with significant anti-HBV activity,and its mechanism of anti-HBV is different from lamivudine.The combination of lamivudine and protocatechuic acid has a positive effect in the treatment of HBV.Gallic acid is very similar to protocatechuic acid in chemical structure and anti-HBV activity.In addition,its bioavailability is limited due to molecule polar and other reasons.Then,the modification of gallic acid is also necessary.In our research group we have found that the combination of lamivudine and protocatechuic acid inhibited the secretion of HBV antigen and HBV DNA replication in Hep G2.2.15 cells,and also inhibited the transcriptional activity of HBV promoter.The result showed these conjugates exhibited better inhibition than lamivudine or protocatechuic acid monotherapy.In this project,lamivudine-protocatechuic acid and lamivudine-gallate acid synergistic prodrugs were designed and synthesized by linking lamivudine with polyphenols by chemical bonding,and the synthesis route and reaction conditions were optimized.Amplification reactions were also conducted on the target compound,and compound 2-2(0.98g),compound 2-3(0.44 g),compound 3-2(50.1 mg),and compound 3-3(101.3 mg)were obtained.The HBV DNA inhibition rate of lamivudine-protocatechuic acid prodrugs 2-2 and2-3 was detected by q RT-PCR method,and the inhibition of these prodrugs on HBe Ag and HBs Ag was also detected by ELISA kit method.The results showed that prodrugs 2-2 and 2-3still retained the anti-HBV activity of lamivudine in vitro,meanwhile there was no significant difference with the lamivudine control group in the inhibitory effect on HBe Ag,HBs Ag and HBV DNA.The other prodrugs will be synthesized and the next step in vivo experiments are still needed in the future.