Hepatitis C Viral Entry Inhibitors Prolong Viral Suppression by Replication Inhibitors in Persistently-Infected Huh7 Cultures

Efforts to treat HCV patients are focused on developing antiviral combinations that lead to the eradication of infection. Thus, it is important to identify optimal combinations from the various viral inhibitor classes. Based on viral dynamic models, HCV entry inhibitors are predicted to reduce viral load in a monophasic manner reflecting the slow death rate of infected hepatocytes (t(1/2) = 2-70 days) and the protection of naive, un-infected cells from HCV infection. In contrast, replication inhibitors are predicted to reduce viral load in a biphasic manner. The initial rapid reduction phase is due to the inhibition of virus production and elimination of plasma virus (t(1/2)similar to 3 hours). The second, slower reduction phase results from the elimination of infected hepatocytes. Here we sought to compare the ability of HCV entry and replication inhibitors as well as combinations thereof to reduce HCV infection in persistently-infected Huh7 cells. Treatment with 5xEC(50) of entry inhibitors anti-CD81 Ab or EI-1 resulted in modest (<= 1 log(10) RNA copies/ml), monophasic declines in viral levels during 3 weeks of treatment. In contrast, treatment with 5xEC(50) of the replication inhibitors BILN-2016 or BMS-790052 reduced extracellular virus levels more potently (similar to 2 log(10) RNA copies/ml) over time in a biphasic manner. However, this was followed by a slow rise to steady-state virus levels due to the emergence of resistance mutations. Combining an entry inhibitor with a replication inhibitor did not substantially enhance the rate of virus reduction. However, entry/replication inhibitor and replication/replication inhibitor combinations reduced viral levels further than monotherapies (up to 3 log(10) RNA copies/ml) and prolonged this reduction relative to monotherapies. Our results demonstrated that HCV entry inhibitors combined with replication inhibitors can prolong antiviral suppression, likely due to the delay of viral resistance emergence.