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Recent HIV\u20101 studies suggest that combination therapies using nucleo(s)tide reverse transcriptase inhibitors (NRTIs) and non\u2010nucleo(s)tide reverse transcriptase inhibitors (NNRTIs) could drastically inhibit the viral genome replication of NRTI\u2010resistant viruses. In order to carry out such combinational therapy against HBV, several new NRTIs and NNRTIs should be developed. Here, we aimed to identify novel NNRTIs targeting the HBV polymerase terminal protein (TP)\u2010reverse transcriptase (RT) (TP\u2010RT) domain, which is a critical domain for HBV replication. We expressed and purified the HBV TP\u2010RT with high purity using an <jats:italic>Escherichia coli<\/jats:italic> expression system and established an in vitro \u03b5 RNA\u2010binding assay system. Then, we used TP\u2010RT in cell\u2010free assays to screen candidate inhibitors from a chemical compound library, and identified two compounds, 6\u2010hydroxy\u2010DL\u2010DOPA and N\u2010oleoyldopamine, which inhibited the binding of \u03b5 RNA with the HBV polymerase. Furthermore, these drugs reduced HBV DNA levels in cell\u2010based assays as well by inhibiting packaging of pregenome RNA into capsids. The novel screening system developed herein should open a new pathway the discovery of drugs targeting the HBV TP\u2010RT domain to treat HBV infection.<\/jats:p>","DOI":"10.1111\/gtc.12778","type":"journal-article","created":{"date-parts":[[2020,5,16]],"date-time":"2020-05-16T15:13:25Z","timestamp":1589642005000},"page":"523-537","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Establishment of a system for finding inhibitors of \u03b5 RNA binding with the HBV polymerase"],"prefix":"10.1111","volume":"25","author":[{"given":"Xiao\u2010Quan","family":"Liu","sequence":"first","affiliation":[{"name":"Division of Virology Department of Microbiology and Immunology Osaka University Graduate School of Medicine Suita Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eriko","family":"Ohsaki","sequence":"additional","affiliation":[{"name":"Division of Virology Department of Microbiology and Immunology Osaka University Graduate School of Medicine Suita Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5641-8105","authenticated-orcid":false,"given":"Keiji","family":"Ueda","sequence":"additional","affiliation":[{"name":"Division of Virology Department of Microbiology and Immunology Osaka University Graduate School of Medicine Suita Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,6,8]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1992.tb05420.x"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M301069200"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v13.i1.48"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2015.10.003"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1254082"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M211231200"},{"issue":"4","key":"e_1_2_7_8_1","first-page":"464","article-title":"Unveiling the roles of HBV polymerase for new antiviral strategies (vol 10, pg 283, 2015)","volume":"10","author":"Clark D. 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