A new strategy to identify hepatitis B virus entry inhibitors by AlphaScreen technology targeting the envelope-receptor interaction

作者：Saso Wakana; Tsukuda Senko; Ohashi Hirofumi; Fukano Kento; Morishita Ryo; Matsunaga Satoko; Ohki Mio; Ryo Akihide; Park Sam Yong; Suzuki Ryosuke; Aizaki Hideki; Muramatsu Masamichi; Sureau Camille; Wakita Takaji; Matano Tetsuro; Watashi Koichi^*

来源：Biochemical and Biophysical Research Communications, 2018, 501(2): 374-379.

DOI：10.1016/j.bbrc.2018.04.187

• HBV
• AlphaScreen
• NTCP
• Rapamycin
• Entry
• Screening

Current anti-hepatitis B virus (HBV) agents have limited effect in curing HBV infection, and thus novel anti-HBV agents with different modes of action are in demand. In this study, we applied AlphaScreen assay to high-throughput screening of small molecules inhibiting the interaction between HBV large surface antigen (LHBs) and the HBV entry receptor, sodium taurocholate cotransporting polypeptide (NTCP). From the chemical screening, we identified that rapamycin, an immunosuppressant, strongly inhibited the LHBs-NTCP interaction. Rapamycin inhibited hepatocyte infection with HBV without significant cytotoxicity. This activity was due to impaired attachment of the LHBs preS1 domain to cell surface. Pretreatment of target cells with rapamycin remarkably reduced their susceptibility to preS1 attachment, while rapamycin pretreatment to preS1 did not affect its attachment activity, suggesting that rapamycin targets the host side. In support of this, a surface plasmon resonance analysis showed a direct interaction of rapamycin with NTCP. Consistently, rapamycin also prevented hepatitis D virus infection, whose entry into cells is also mediated by NTCP. We also identified two rapamycin derivatives, everolimus and temsirolimus, which possessed higher anti-HBV potencies than rapamycin. Thus, this is the first report for application of AlphaScreen technology that monitors a viral envelope-receptor interaction to identify viral entry inhibitors.

• 出版日期2018-6-22
• 单位河北医科大学