Artificial recombinant cell-penetrating peptides interfere with envelopment of hepatitis B virus nucleocapsid and viral production.

Hepatitis B virus (HBV) is a major human infectious pathogen, with over 300 million chronically infected patients worldwide. Current therapeutics for chronic HBV infection have shown only limited success. The plasma membrane represents an impermeable barrier for development of most macromolecular antiviral agents. To develop new anti-HBV macromolecules that can cross the membrane barrier, we designed a series of artificial recombinant peptides including cell penetrating sequence oligoarginine R7 and several nucleocapsid binding subunits (NBS). The anti-HBV function of these peptides was evaluated in a HBV DNA replicative cell line HepG2.2.15. Our results showed that the synthetic recombinant cell penetrating peptides retained the activity of cell penetrating in the living cells. HBV DNA in culture medium markedly decreased in cells treated with cell penetrating peptides bearing NBS for three days. Intracellular HBcAg and HBV DNA replicative intermediates increased by 2-3 fold. In conclusion, the synthetic recombinant cell penetrating peptides bearing NBS can efficiently enter into the cells; block nucleocapsid assembly and inhibit HBV release. Cell penetrating subunit presents a high efficiency tool to deliver synthetic antiviral peptides into cells.