{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T15:37:18Z","timestamp":1790869038159,"version":"4.1.0"},"reference-count":64,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,7,6]],"date-time":"2020-07-06T00:00:00Z","timestamp":1593993600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Sardinian Regional Government","award":["F76C18000800002"],"award-info":[{"award-number":["F76C18000800002"]}]},{"name":"Spanish Ministerio de Ciencia e Innovaci\u00f3n","award":["-"],"award-info":[{"award-number":["-"]}]},{"DOI":"10.13039\/501100003407","name":"Ministero dell\u2019Istruzione, dell\u2019Universit\u00e0 e della Ricerca","doi-asserted-by":"publisher","award":["PRIN 2017BMK8JR"],"award-info":[{"award-number":["PRIN 2017BMK8JR"]}],"id":[{"id":"10.13039\/501100003407","id-type":"DOI","asserted-by":"publisher"}]},{"name":"ISAR Chu Family Foundation Scholarships for Early Career Women in Science","award":["-"],"award-info":[{"award-number":["-"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>HIV-1 infection requires life-long treatment and with 2.1 million new infections\/year, faces the challenge of an increased rate of transmitted drug-resistant mutations. Therefore, a constant and timely effort is needed to identify new HIV-1 inhibitors active against drug-resistant variants. The ribonuclease H (RNase H) activity of HIV-1 reverse transcriptase (RT) is a very promising target, but to date, still lacks an efficient inhibitor. Here, we characterize the mode of action of N\u2019-(2-hydroxy-benzylidene)-3,4,5-trihydroxybenzoylhydrazone (compound 13), an N-acylhydrazone derivative that inhibited viral replication (EC50 = 10 \u00b5M), while retaining full potency against the NNRTI-resistant double mutant K103N-Y181C virus. Time-of-addition and biochemical assays showed that compound 13 targeted the reverse-transcription step in cell-based assays and inhibited the RT-associated RNase H function, being &gt;20-fold less potent against the RT polymerase activity. Docking calculations revealed that compound 13 binds within the RNase H domain in a position different from other selective RNase H inhibitors; site-directed mutagenesis studies revealed interactions with conserved amino acid within the RNase H domain, suggesting that compound 13 can be taken as starting point to generate a new series of more potent RNase H selective inhibitors active against circulating drug-resistant variants.<\/jats:p>","DOI":"10.3390\/v12070729","type":"journal-article","created":{"date-parts":[[2020,7,6]],"date-time":"2020-07-06T11:07:42Z","timestamp":1594033662000},"page":"729","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Targeting HIV-1 RNase H: N\u2019-(2-Hydroxy-benzylidene)-3,4,5-Trihydroxybenzoylhydrazone as Selective Inhibitor Active against NNRTIs-Resistant Variants"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6630-8636","authenticated-orcid":false,"given":"Angela","family":"Corona","sequence":"first","affiliation":[{"name":"Department of Life and Environmental Sciences University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ester","family":"Ballana","sequence":"additional","affiliation":[{"name":"AIDS Research Institute\u2014IrsiCaixa, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simona","family":"Distinto","sequence":"additional","affiliation":[{"name":"Department of Life and Environmental Sciences University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2295-5783","authenticated-orcid":false,"given":"Dominga","family":"Rogolino","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Del Vecchio","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5888-4556","authenticated-orcid":false,"given":"Mauro","family":"Carcelli","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, 43124 Parma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger","family":"Badia","sequence":"additional","affiliation":[{"name":"AIDS Research Institute\u2014IrsiCaixa, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Riveira-Mu\u00f1oz","sequence":"additional","affiliation":[{"name":"AIDS Research Institute\u2014IrsiCaixa, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Esposito","sequence":"additional","affiliation":[{"name":"Department of Life and Environmental Sciences University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Parolin","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padova, 35122 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1436-5823","authenticated-orcid":false,"given":"Jos\u00e9 A.","family":"Est\u00e9","sequence":"additional","affiliation":[{"name":"AIDS Research Institute\u2014IrsiCaixa, 08916 Badalona, Spain"},{"name":"CienciaTraducida, 08391 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Grandi","sequence":"additional","affiliation":[{"name":"Department of Life and Environmental Sciences University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4849-0980","authenticated-orcid":false,"given":"Enzo","family":"Tramontano","sequence":"additional","affiliation":[{"name":"Department of Life and Environmental Sciences University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, Italy"},{"name":"Istituto di Ricerca Genetica e Biomedica, Consiglio Nazionale delle Ricerche (CNR), 09042 Monserrato, Cagliari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,6]]},"reference":[{"key":"ref_1","unstructured":"(2020, February 17). UNAIDS 2018 Global HIV Statistics. Available online: https:\/\/www.unaids.org\/sites\/default\/files\/media_asset\/UNAIDS_FactSheet_en.pdf."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1097\/QAI.0000000000002275","article-title":"Durable Efficacy of Dolutegravir Plus Lamivudine in Antiretroviral Treatment-Naive Adults With HIV-1 Infection: 96-Week Results From the GEMINI-1 and GEMINI-2 Randomized Clinical Trials","volume":"83","author":"Cahn","year":"2020","journal-title":"J. Acquir. Immune Defic. Syndr."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1001\/jama.2012.8142","article-title":"Toward an AIDS-free generation","volume":"308","author":"Fauci","year":"2012","journal-title":"JAMA J. Am. Med. Assoc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1525","DOI":"10.1016\/S0140-6736(13)61809-7","article-title":"The end of AIDS: HIV infection as a chronic disease","volume":"382","author":"Deeks","year":"2013","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1126\/science.367.6478.611","article-title":"Combo of two HIV vaccines fails its big test","volume":"367","author":"Cohen","year":"2020","journal-title":"Science"},{"key":"ref_6","first-page":"1","article-title":"Advances toward Curing HIV-1 Infection in Tissue Reservoirs","volume":"94","author":"Henderson","year":"2019","journal-title":"J. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"S847","DOI":"10.1093\/infdis\/jix430","article-title":"The Role of HIV-1 Drug-Resistant Minority Variants in Treatment Failure","volume":"216","author":"Arribas","year":"2017","journal-title":"J. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1016\/S1473-3099(17)30702-8","article-title":"HIV-1 drug resistance before initiation or re-initiation of first-line antiretroviral therapy in low-income and middle-income countries: A systematic review and meta-regression analysis","volume":"18","author":"Gupta","year":"2017","journal-title":"Lancet Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S1473-3099(15)00536-8","article-title":"Global epidemiology of drug resistance after failure of WHO recommended first-line regimens for adult HIV-1 infection: A multicentre retrospective cohort study","volume":"16","author":"Gregson","year":"2016","journal-title":"Lancet Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2310","DOI":"10.1093\/nar\/gkw060","article-title":"Biochemical characterization of a multi-drug resistant HIV-1 subtype AG reverse transcriptase: Antagonism of AZT discrimination and excision pathways and sensitivity to RNase H inhibitors","volume":"44","author":"Schneider","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_11","unstructured":"Schatz, O., Cromme, F.V., Naas, T., and Lindemann, D. (1990). Inactivation of the RNase H domain of HIV-1 reverse transcriptase blocks viral infectivity. Gene Regulation and AIDS, Gulf Publishing Company."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1126\/science.1707186","article-title":"Crystal structure of the ribonuclease H domain of HIV-1 reverse transcriptase","volume":"252","author":"Davies","year":"1991","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/JVI.00515-18","article-title":"Ortervirales: New Virus Order Unifying Five Families of Reverse-Transcribing Viruses","volume":"92","author":"Krupovic","year":"2018","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1111\/j.1742-4658.2009.06907.x","article-title":"Ribonuclease H: Molecular diversities, substrate binding domains, and catalytic mechanism of the prokaryotic enzymes","volume":"276","author":"Tadokoro","year":"2009","journal-title":"FEBS J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/microbiolspec.MDNA3-0024-2014","article-title":"Retroviral integrase structure and dna recombination mechanism","volume":"2","author":"Engelman","year":"2014","journal-title":"Microbiol. Spectr."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"102672","DOI":"10.1016\/j.dnarep.2019.102672","article-title":"RNases H: Structure and mechanism","volume":"84","author":"Hyjek","year":"2019","journal-title":"DNA Repair."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4086","DOI":"10.1128\/AAC.00056-14","article-title":"Inhibition of foamy virus reverse transcriptase by human immunodeficiency virus type 1 ribonuclease H inhibitors","volume":"58","author":"Corona","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.4155\/fmc.13.178","article-title":"Active site and allosteric inhibitors of the ribonuclease H activity of HIV reverse transcriptase","volume":"5","author":"Corona","year":"2013","journal-title":"Futur. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1682","DOI":"10.2174\/0929867324666170113110839","article-title":"Update on Recent Developments in Small Molecular HIV-1 RNase H Inhibitors (2013\u20132016): Opportunities and Challenges","volume":"25","author":"Wang","year":"2018","journal-title":"Curr. Med. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Tramontano, E., Corona, A., and Men\u00e9ndez-Arias, L. (2019). Ribonuclease H, an unexploited target for antiviral intervention against HIV and hepatitis B virus. Antivir. Res., 171.","DOI":"10.1016\/j.antiviral.2019.104613"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"727","DOI":"10.2174\/138955706777435733","article-title":"HIV-1 RNase H: Recent progress in an exciting, yet little explored, drug target","volume":"6","author":"Tramontano","year":"2006","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1016\/j.ejmech.2018.07.039","article-title":"6-Arylthio-3-hydroxypyrimidine-2,4-diones potently inhibited HIV reverse transcriptase-associated RNase H with antiviral activity","volume":"156","author":"Wang","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1963","DOI":"10.1021\/acsinfecdis.9b00300","article-title":"Determinants of active-site inhibitor interaction with HIV-1 RNase H","volume":"5","author":"Xi","year":"2019","journal-title":"ACS Infect. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"445","DOI":"10.2217\/fvl.14.24","article-title":"Can the ever-promising target HIV reverse transcriptase-associated RNase H become a success story for drug development?","volume":"9","author":"Corona","year":"2014","journal-title":"Future Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1021\/acs.accounts.9b00450","article-title":"Cutting into the Substrate Dominance: Pharmacophore and Structure-Based Approaches toward Inhibiting Human Immunodeficiency Virus Reverse Transcriptase-Associated Ribonuclease H","volume":"53","author":"Wang","year":"2020","journal-title":"Acc. Chem. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6101","DOI":"10.1128\/AAC.03605-14","article-title":"Identification of Highly Conserved Residues Involved in Inhibition of HIV-1 RNase H Function by Diketo Acid Derivatives","volume":"58","author":"Corona","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1039\/C7MD00600D","article-title":"Structure-guided approach identifies a novel class of HIV-1 ribonuclease H inhibitors: Binding mode insights through magnesium complexation and site-directed mutagenesis studies","volume":"9","author":"Poongavanam","year":"2018","journal-title":"Medchemcomm"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"798","DOI":"10.1021\/acsmedchemlett.9b00617","article-title":"Pyrrolyl Pyrazoles as Non-Diketo Acid Inhibitors of the HIV-1 Ribonuclease H Function of Reverse Transcriptase","volume":"11","author":"Messore","year":"2020","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.antiviral.2016.09.008","article-title":"New insights into the interaction between pyrrolyl diketoacids and HIV-1 integrase active site and comparison with RNase H","volume":"134","author":"Corona","year":"2016","journal-title":"Antivir. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"440","DOI":"10.3389\/fmicb.2017.00440","article-title":"Chelation motifs affecting metal-dependent viral enzymes: N-acylhydrazone ligands as dual target inhibitors of HIV-1 integrase and reverse transcriptase ribonuclease Hdomain","volume":"8","author":"Carcelli","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4856","DOI":"10.1021\/bi015970t","article-title":"Altering the RNase H primer grip of human immunodeficiency virus reverse transcriptase modifies cleavage specificity","volume":"41","author":"Rausch","year":"2002","journal-title":"Biochemistry"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"9301","DOI":"10.1128\/JVI.01006-10","article-title":"Exploiting Drug Repositioning for Discovery of a Novel HIV Combination Therapy","volume":"84","author":"Clouser","year":"2010","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1038\/nprot.2007.517","article-title":"Tetrazolium-based colorimetric assay for the detection of HIV replication inhibitors: Revisited 20 years later","volume":"3","author":"Pannecouque","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.antiviral.2015.09.004","article-title":"The thioacetate-\u03c9(\u03b3-lactam carboxamide) HDAC inhibitor ST7612AA1 as HIV-1 latency reactivation agent","volume":"123","author":"Badia","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1038\/nprot.2011.330","article-title":"A time-of-drug addition approach to target identification of antiviral compounds","volume":"6","author":"Daelemans","year":"2011","journal-title":"Nat. Protoc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1002\/jcc.540110405","article-title":"Macromodel\u2014An integrated software system for modeling organic and bioorganic molecules using molecular mechanics","volume":"11","author":"Mohamadi","year":"1990","journal-title":"J. Comput. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1002\/(SICI)1096-987X(199604)17:5\/6<520::AID-JCC2>3.0.CO;2-W","article-title":"Merck molecular force field. II. MMFF94 van der Waals and electrostatic parameters for intermolecular interactions","volume":"17","author":"Halgren","year":"1996","journal-title":"J. Comput. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6127","DOI":"10.1021\/ja00172a038","article-title":"Semianalytical treatment of solvation for molecular mechanics and dynamics","volume":"112","author":"Still","year":"1990","journal-title":"J. Am. Chem. Soc."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7625","DOI":"10.1128\/JVI.00353-10","article-title":"Structural basis for the inhibition of RNase H activity of HIV-1 reverse transcriptase by RNase H active site-directed inhibitors","volume":"84","author":"Su","year":"2010","journal-title":"J. Virol."},{"key":"ref_41","unstructured":"Schr\u00f6dinger LLC (2013). Maestro GUI, Schr\u00f6dinger LLC."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"11225","DOI":"10.1021\/ja9621760","article-title":"Development and Testing of the OLPS All-Atom Force Field on Conformational Energetics and Properties of Organic Liquids","volume":"118","author":"Jorgensen","year":"1996","journal-title":"J. Am. Chem. Soc."},{"key":"ref_43","unstructured":"Schr\u00f6dinger LLC QMPolarized Protocol, Schr\u00f6dinger LLC."},{"key":"ref_44","unstructured":"Schr\u00f6dinger LLC The PyMOL Molecular Graphics System, Version 1.7.2.1 2014, Schr\u00f6dinger LLC."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1021\/ci049885e","article-title":"LigandScout: 3-D pharmacophores derived from protein-bound ligands and their use as virtual screening filters","volume":"45","author":"Wolber","year":"2005","journal-title":"J. Chem. Inf. Model."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1007\/s10822-006-9078-7","article-title":"Efficient Overlay of Small Organic Molecules Using 3D Pharmacophores","volume":"20","author":"Wolber","year":"2006","journal-title":"J. Comput. Aided. Mol. Des."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Corona, A., Meleddu, R., Esposito, F., Distinto, S., Bianco, G., Masaoka, T., Maccioni, E., Men\u00e9ndez-Arias, L., Alcaro, S., and Le Grice, S.F.J. (2016). Ribonuclease H\/DNA Polymerase HIV-1 Reverse Transcriptase Dual Inhibitor: Mechanistic Studies on the Allosteric Mode of Action of Isatin-Based Compound RMNC6. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0147225"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Pala, N., Esposito, F., Rogolino, D., Carcelli, M., Sanna, V., Palomba, M., Naesens, L., Corona, A., Grandi, N., and Tramontano, E. (2016). Inhibitory effect of 2,3,5,6-tetrafluoro-4-[4-(Aryl)-1H-1,2,3-triazol-1-yl]benzenesulfonamide derivatives on HIV reverse transcriptase associated RNase H activities. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17081371"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejmech.2018.10.029","article-title":"Novel natural non-nucleoside inhibitors of HIV-1 reverse transcriptase identified by shape- and structure-based virtual screening techniques","volume":"161","author":"Costa","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1097\/01.aids.0000216363.36786.2b","article-title":"HIV-1 pol mutation frequency by subtype and treatment experience: Extension of the HIVseq program to seven non-B subtypes","volume":"20","author":"Rhee","year":"2006","journal-title":"AIDS"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1093\/nar\/gkg100","article-title":"Human immunodeficiency virus reverse transcriptase and protease sequence database","volume":"31","author":"Rhee","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1647","DOI":"10.1093\/bioinformatics\/bts199","article-title":"Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data","volume":"28","author":"Kearse","year":"2012","journal-title":"Bioinformatics"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT multiple sequence alignment software version 7: Improvements in performance and usability","volume":"30","author":"Katoh","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1850","DOI":"10.2174\/1381612811319100009","article-title":"Molecular Aspects of the RT\/drug Interactions. Perspective of Dual Inhibitors","volume":"19","author":"Distinto","year":"2013","journal-title":"Curr. Pharm. Des."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1016\/j.str.2009.09.016","article-title":"Structure of HIV-1 reverse transcriptase with the inhibitor beta-Thujaplicinol bound at the RNase H active site","volume":"17","author":"Himmel","year":"2009","journal-title":"Structure"},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Kirby, K.A., Myshakina, N.A., Christen, M.T., Chen, Y.L., Schmidt, H.A., Huber, A.D., Xi, Z., Kim, S., Rao, R.K., and Kramer, S.T. (2017). A 2-hydroxyisoquinoline-1,3-dione active-site RNase H inhibitor binds in multiple modes to HIV-1 reverse transcriptase. Antimicrob. Agents Chemother., 61.","DOI":"10.1128\/AAC.01351-17"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.antiviral.2004.11.002","article-title":"6-[1-(4-Fluorophenyl)methyl-1H-pyrrol-2-yl)]-2,4-dioxo-5-hexenoic acid ethyl ester a novel diketo acid derivative which selectively inhibits the HIV-1 viral replication in cell culture and the ribonuclease H activity in vitro","volume":"65","author":"Tramontano","year":"2005","journal-title":"Antivir. Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/JVI.02203-17","article-title":"Developing and Evaluating Inhibitors against the RNase H Active Site of HIV-1 Reverse Transcriptase","volume":"92","author":"Boyer","year":"2018","journal-title":"J. Virol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1074\/jbc.M110254200","article-title":"Mutational analysis of Tyr-501 of HIV-1 reverse transcriptase. Effects on ribonuclease H activity and inhibition of this activity by N-acylhydrazones","volume":"277","author":"Arion","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1124\/mol.62.2.398","article-title":"Destabilization of the HIV-1 Reverse Transcriptase Dimer upon Interaction with N-Acyl Hydrazone Inhibitors","volume":"62","year":"2002","journal-title":"Mol. Pharmacol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1111\/cbdd.13455","article-title":"Design, synthesis, and biologic evaluation of novel galloyl derivatives as HIV-1 RNase H inhibitors","volume":"93","author":"Gao","year":"2019","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1016\/j.ejmech.2018.07.035","article-title":"6-Biphenylmethyl-3-hydroxypyrimidine-2,4-diones potently and selectively inhibited HIV reverse transcriptase-associated RNase H","volume":"156","author":"Wang","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1709","DOI":"10.1002\/cmdc.201600015","article-title":"Studies on Cycloheptathiophene-3-carboxamide Derivatives as Allosteric HIV-1 Ribonuclease H Inhibitors","volume":"11","author":"Corona","year":"2016","journal-title":"ChemMedChem"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"6852","DOI":"10.1093\/nar\/gkg881","article-title":"Two-metal ion mechanism of RNA cleavage by HIV RNase H and mechanism-based design of selective HIV RNase H inhibitors","volume":"31","author":"Klumpp","year":"2003","journal-title":"Nucleic Acids Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/7\/729\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:47:56Z","timestamp":1760176076000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/7\/729"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,6]]},"references-count":64,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["v12070729"],"URL":"https:\/\/doi.org\/10.3390\/v12070729","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,6]]}}}