{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T13:57:28Z","timestamp":1790863048785,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82572621"],"award-info":[{"award-number":["82572621"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Sanming Project of Medicine in Shenzen Municipality","award":["SMZ202303037"],"award-info":[{"award-number":["SMZ202303037"]}]},{"name":"Shenzhen Key Medical Discipline Construction Found","award":["SZXK06162"],"award-info":[{"award-number":["SZXK06162"]}]},{"name":"Science, Technology and Innovation Commission of Basic research funds","award":["JCYJ20240813114518024, KJZD20240903103500002, JCYJ20240813114540052"],"award-info":[{"award-number":["JCYJ20240813114518024, KJZD20240903103500002, JCYJ20240813114540052"]}]},{"name":"Shenzhen Nanshan District Scientific Research Program of People's Republic of China","award":["NS2024044Y, NS2024006Y, NS2024007Y, NSZD2024036, NSZD2024023, NSZD2024032, NSZD2025001, NSZD2025005, NSZD2025006"],"award-info":[{"award-number":["NS2024044Y, NS2024006Y, NS2024007Y, NSZD2024036, NSZD2024023, NSZD2024032, NSZD2025001, NSZD2025005, NSZD2025006"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Staphylococcus aureus<\/jats:italic>\n                      infections pose a significant threat to global human health, particularly with the widespread transmission of methicillin-resistant\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      (MRSA) strains, which has created an urgent need for the development of new antimicrobial agents. PFM01 is an N-alkylated derivative of Mirin, an MRE11 endonuclease inhibitor. It was initially discovered to block the repair of double-strand DNA breaks and demonstrated preliminary anti-tumor potential; however, its antimicrobial activity had not yet been explored. Here, the anti-\n                      <jats:italic toggle=\"yes\">S<\/jats:italic>\n                      .\n                      <jats:italic toggle=\"yes\">aureus<\/jats:italic>\n                      activity of PFM01 was determined by broth microdilution minimum inhibitory concentration (MIC) testing and time-kill kinetics. The\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      cytotoxicity was assessed by CCK-8 assay and hemolysis test. The potential mechanisms were systematically investigated by whole-genome sequencing, drug affinity responsive target stability (DARTS), and molecular docking analyses. PFM01 exhibited potent antibacterial activity against Gram-positive bacteria (including\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Enterococcus faecalis<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">Enterococcus faecium<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">Streptococcus lactis<\/jats:italic>\n                      ), with MIC\n                      <jats:sub>50<\/jats:sub>\n                      values ranging from 6.25 to 12.5 \u03bcM (1.83\u20133.67 \u03bcg\/mL). Time-kill curve analysis confirmed its bactericidal capacity against free-floating\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      (including bacteria in the logarithmic growth phase and stationary phase-retained cells). Furthermore, PFM01 inhibited\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      biofilm formation in a concentration-dependent manner and possesses the ability to remove mature biofilms.\n                      <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                      cellular safety tests of PFM01 indicated low cytotoxicity and no significant hemolytic activity. Moreover, PFM01 altered the membrane permeability and potential of\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      , and its antibacterial activity was counteracted by the exogenous phospholipids. Whole-genome sequencing identified genetic mutations in the\n                      <jats:italic toggle=\"yes\">rapZ<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">mupR<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">mprF,<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">clpX<\/jats:italic>\n                      genes of PFM01-resistant\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      strains; however, the antibacterial activity of PFM01 against\n                      <jats:italic toggle=\"yes\">mprF<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">clpX<\/jats:italic>\n                      knockout\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      strains showed no significant change compared with wild-type strains. Proteomic analysis further revealed that PFM01 primarily might interfere with purine metabolism, amino acid biosynthesis, and the phosphotransferase system. DARTS experiments suggested that glycerol-3-phosphate cytidylyltransferase (TarD) might be a potential target, and molecular docking results further confirmed a stable interaction between PFM01 and TarD (binding energy: \u22127.4 kcal\/mol). In summary, PFM01 may exert its antibacterial effect by disrupting the bacterial cell membrane structure and targeting TarD, and holds promise as a therapeutic agent for\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      infections.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        This study confirms that PFM01 has a broad and strong antibacterial effect against Gram-positive bacteria while also being biologically safe. The antimicrobial mechanism of PFM01 is closely associated with TarD. The advantages of low cytotoxicity against host cells and reduced hemolysis toward red cells were further demonstrated. As a promising repurposed antimicrobial candidate, PFM01 shows potential application for the treatment of infections caused by multidrug-resistant\n                        <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                        .\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01372-26","type":"journal-article","created":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T13:03:08Z","timestamp":1790859788000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Repurposing PFM01 as a novel antibacterial agent against\n                    <i>Staphylococcus aureus<\/i>"],"prefix":"10.1128","author":[{"given":"Congcong","family":"Li","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01vy4gh70","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Basic Medical Sciences, Shenzhen University 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