{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T04:40:45Z","timestamp":1767069645504,"version":"build-2065373602"},"reference-count":68,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2025,1,26]],"date-time":"2025-01-26T00:00:00Z","timestamp":1737849600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-19-AMRB-0001"],"award-info":[{"award-number":["ANR-19-AMRB-0001"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemMedChem"],"published-print":{"date-parts":[[2025,4,14]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Aminoglycoside\u2010phosphotransferases (APHs) are a class of bacterial enzymes that mediate acquired resistance to aminoglycoside antibiotics. Here we report the identification of small molecules counteracting aminoglycoside resistance in <jats:italic>Enterococcus casseliflavus<\/jats:italic>. Molecular dynamics simulations were performed to identify an allosteric pocket in three APH enzymes belonging to 3\u2032 and 2\u2032\u2032 subfamilies in which we then screened, <jats:italic>in silico<\/jats:italic>, 12,000 small molecules. From a subset of only 14 high\u2010scored molecules tested <jats:italic>in\u2005vitro<\/jats:italic>, we identified a compound, named here <jats:bold>EK3<\/jats:bold>, able to non\u2010competitively inhibit the APH(2\u2032\u2032)\u2010IVa, an enzyme mediating clinical gentamicin resistance. Structure\u2010activity relationship (SAR) exploration of this hit compound allowed us to identify a molecule with improved enzymatic inhibition. By measuring bacterial sensitivity, we found that the three best compounds in this series restored bactericidal activity of various aminoglycosides, including gentamicin, without exhibiting toxicity to HeLa cells. This work not only provides a basis to fight aminoglycoside resistance but also highlights a proof\u2010of\u2010concept for the search of allosteric modulators by using <jats:italic>in silico<\/jats:italic> methods.<\/jats:p>","DOI":"10.1002\/cmdc.202400842","type":"journal-article","created":{"date-parts":[[2025,1,13]],"date-time":"2025-01-13T04:54:14Z","timestamp":1736744054000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["APH Inhibitors that Reverse Aminoglycoside Resistance in <i>Enterococcus casseliflavus<\/i>"],"prefix":"10.1002","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4985-1482","authenticated-orcid":false,"given":"Elise","family":"Kaplan","sequence":"first","affiliation":[{"name":"Institut de Recherche en Infectiologie de Montpellier \u2013 IRIM University of Montpellier CNRS UMR 9004  1919 route de Mende 34293 Montpellier cedex 5 France"},{"name":"Current address: University of Lyon CNRS, UMR5086, Molecular Microbiology and Structural Biochemistry, IBCP  7 Passage du Vercors 69367 Lyon France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5757-5804","authenticated-orcid":false,"given":"Laurent","family":"Chaloin","sequence":"additional","affiliation":[{"name":"Institut de Recherche en Infectiologie de Montpellier \u2013 IRIM University of Montpellier CNRS UMR 9004  1919 route de Mende 34293 Montpellier cedex 5 France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7699-3235","authenticated-orcid":false,"given":"Jean\u2010Fran\u00e7ois","family":"Guichou","sequence":"additional","affiliation":[{"name":"Centre de Biologie Structurale \u2013 CBS University of Montpellier CNRS UMR 5048 INSERM U 1054  29 rue de Navacelles 34090 Montpellier France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2295-2702","authenticated-orcid":false,"given":"K\u00e9vin","family":"Berrou","sequence":"additional","affiliation":[{"name":"Institut de Recherche en Infectiologie de Montpellier \u2013 IRIM University of Montpellier CNRS UMR 9004  1919 route de Mende 34293 Montpellier cedex 5 France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4992-842X","authenticated-orcid":false,"given":"Rahila","family":"Rahimova","sequence":"additional","affiliation":[{"name":"Centre de Biologie Structurale \u2013 CBS University of Montpellier CNRS UMR 5048 INSERM U 1054  29 rue de Navacelles 34090 Montpellier France"},{"name":"Current address: University of Grenoble Alpes, CEA, CNRS, IBS, Metalloproteins Unit, 71 avenue des Martyrs  CS 10090, 38000 Grenoble France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6861-3300","authenticated-orcid":false,"given":"Gilles","family":"Labesse","sequence":"additional","affiliation":[{"name":"Centre de Biologie Structurale \u2013 CBS University of Montpellier CNRS UMR 5048 INSERM U 1054  29 rue de Navacelles 34090 Montpellier France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6348-5311","authenticated-orcid":false,"given":"Corinne","family":"Lionne","sequence":"additional","affiliation":[{"name":"Institut de Recherche en Infectiologie de Montpellier \u2013 IRIM University of Montpellier CNRS UMR 9004  1919 route de Mende 34293 Montpellier cedex 5 France"},{"name":"Centre de Biologie Structurale \u2013 CBS University of Montpellier CNRS UMR 5048 INSERM U 1054  29 rue de Navacelles 34090 Montpellier France"},{"name":"Current address: University of Montpellier, CNRS UMR 5048, INSERM U 1054, CBS, 29 rue de Navacelles  34090 Montpellier France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,1,26]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a027029"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.12.3249-3256.2000"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/35030019"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/mr.51.3.341-350.1987"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1177\/106002809102500210"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.1.2.139"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.47.1.144-147.2003"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.drup.2010.08.003"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.16.3.430-450.2003"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(02)00125-4"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/ja.2016.144"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja964278w"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1399004714017635"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80274-3"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja00150a004"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja0543144"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2005.06.059"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00572-10"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.40.24755"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2011.10.018"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20130317"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagen.2016.09.012"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2005.04.020"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00188a024"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/21.10.2323"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi6024512"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.10.2691-2694.2001"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.5.1229"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi010504p"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00149-09"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.349670"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi036095"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M513257200"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2012.10.027"},{"key":"e_1_2_9_35_1","unstructured":"E. Kaplan J.\u2005F. Guichou L. Chaloin S. Kunzelmann N. Leban E.\u2005H. Serpersu C. Lionne Biochim. Biophys. Acta \u2013 Gen. Subj.2016 DOI10.1016\/j.bbagen.2016.01.016."},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1021\/ci049714"},{"key":"e_1_2_9_37_1","unstructured":"J.\u2005J. Irwin T. Sterling M.\u2005M. Mysinger E.\u2005S. Bolstad R.\u2005G. Coleman 2012 DOI10.1021\/ci3001277."},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1996.0897"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.10465"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep10754"},{"key":"e_1_2_9_41_1","unstructured":"T.\u2005E. Barman S.\u2005R.\u2005W. Bellamy H. Gutfreund S.\u2005E. Halford C. Lionne Cell. Mol. Life Sci.2006 DOI10.1007\/s00018-006-6243-z."},{"key":"e_1_2_9_42_1","unstructured":"M. Toth H. Frase N.\u2005T. Antunes C.\u2005A. Smith S.\u2005B. Vakulenko Protein Sci.2010 DOI10.1002\/pro.437."},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13321-015-0069-3"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.437"},{"key":"e_1_2_9_45_1","unstructured":"The European Committee on Antimicrobial Susceptibility Testing.Breakpoint Tables for Interpretation of MICs and Zone Diameters. Version 10.0.2020."},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(01)00051-5"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019390"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0019589"},{"key":"e_1_2_9_49_1","unstructured":"L.\u2005A. Velez S.\u2005D. Arteaga 2024 DOI10.1101\/2024.12.23.630124."},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(63)80091-1"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-5521(00)00115-0"},{"key":"e_1_2_9_52_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1169378"},{"key":"e_1_2_9_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pharmthera.2015.10.002"},{"key":"e_1_2_9_54_1","unstructured":"European Committee for Antimicrobial Susceptibility Testing (EUCAST) of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Clin. Microbiol. Infect.2003 9 ix\u2013xv."},{"key":"e_1_2_9_55_1","doi-asserted-by":"publisher","DOI":"10.1021\/ct300400x"},{"key":"e_1_2_9_56_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20289"},{"key":"e_1_2_9_57_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-10-168"},{"key":"e_1_2_9_58_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkq383"},{"key":"e_1_2_9_59_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btr550"},{"key":"e_1_2_9_60_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkh368"},{"key":"e_1_2_9_61_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagen.2016.09.012"},{"key":"e_1_2_9_62_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444911007773"},{"key":"e_1_2_9_63_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444913000061"},{"key":"e_1_2_9_64_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889807021206"},{"key":"e_1_2_9_65_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"},{"key":"e_1_2_9_66_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444911001314"},{"key":"e_1_2_9_67_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.10286"},{"key":"e_1_2_9_68_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889892009944"}],"container-title":["ChemMedChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cmdc.202400842","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T14:08:04Z","timestamp":1760278084000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cmdc.202400842"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,26]]},"references-count":68,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2025,4,14]]}},"alternative-id":["10.1002\/cmdc.202400842"],"URL":"https:\/\/doi.org\/10.1002\/cmdc.202400842","archive":["Portico"],"relation":{},"ISSN":["1860-7179","1860-7187"],"issn-type":[{"type":"print","value":"1860-7179"},{"type":"electronic","value":"1860-7187"}],"subject":[],"published":{"date-parts":[[2025,1,26]]},"assertion":[{"value":"2024-10-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-01-26","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202400842"}}