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High-Throughput Screening of Antiviral Compounds Using a Recombinant Hepatitis B Virus and Identification of a Possible Infection Inhibitor, Skimmianine

Authors :
Mika Yoshita
Masaya Funaki
Tetsuro Shimakami
Masaki Kakuya
Kazuhisa Murai
Saiho Sugimoto
Shotaro Kawase
Koji Matsumori
Taro Kawane
Tomoki Nishikawa
Asuka Nakamura
Reo Suzuki
Atsuya Ishida
Narumi Kawasaki
Yuga Sato
Ying-Yi Li
Ariunaa Sumiyadorj
Kouki Nio
Hajime Takatori
Kazunori Kawaguchi
Kazuyuki Kuroki
Takanobu Kato
Masao Honda
Taro Yamashita
Source :
Viruses, Vol 16, Iss 8, p 1346 (2024)
Publication Year :
2024
Publisher :
MDPI AG, 2024.

Abstract

We developed a novel hepatitis B virus (HBV) infection-monitoring system using a luminescent, 11-amino acid reporter (HiBiT). We performed high-throughput antiviral screening using this system to identify anti-HBV compounds. After the infection of primary human hepatocytes with the recombinant virus HiBiT-HBV, which contains HiBiT at its preS1, 1262 compounds were tested in a first screening using extracellular HiBiT activity as an indicator of viral infection. Following a second screening, we focused on the compound skimmianine, which showed a potent antiviral effect. When skimmianine was added at the same time as HiBiT-HBV infection, skimmianine inhibited HiBiT activity with EC50 of 0.36 pM, CC50 of 1.67 μM and a selectivity index (CC50:EC50 ratio) of 5,100,000. When skimmianine was added 72 h after HiBiT-HBV infection, the EC50, CC50 and selectivity index were 0.19 μM, 1.87 μM and 8.79, respectively. Time-lapse fluorescence imaging analysis using another recombinant virus, ReAsH-TC155HBV, with the insertion of tetra-cysteine within viral capsid, revealed that skimmianine inhibited the accumulation of the capsid into hepatocytes. Furthermore, skimmianine did not inhibit either attachment or internalization. These results imply that skimmianine inhibits the retrograde trafficking of the virus after internalization. This study demonstrates the usefulness of the recombinant virus, HiBiT-HBV, for high-throughput screening to identify anti-HBV compounds.

Details

Language :
English
ISSN :
19994915
Volume :
16
Issue :
8
Database :
Directory of Open Access Journals
Journal :
Viruses
Publication Type :
Academic Journal
Accession number :
edsdoj.9b0e87ed2448cd98de2ee68561f75c
Document Type :
article
Full Text :
https://doi.org/10.3390/v16081346