{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T22:21:08Z","timestamp":1785536468802,"version":"3.56.0"},"reference-count":34,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005047","name":"Natural Science Foundation of Liaoning Province","doi-asserted-by":"crossref","award":["2025-BS-0858"],"award-info":[{"award-number":["2025-BS-0858"]}],"id":[{"id":"10.13039\/501100005047","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Project of the Science and Technology Department of Liaoning Province","award":["2024-MS-226"],"award-info":[{"award-number":["2024-MS-226"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>To address the urgent challenge of antimicrobial resistance, a series of twenty novel C-28 modified betulinic acid derivatives was designed and synthesized. Several derivatives, particularly 3b, 3d, 3e, and 3o, displayed notable antibacterial activity against Gram-positive bacteria, including Staphylococcus aureus and vancomycin-resistant Staphylococcus aureus (VRSA). The most active compound, 3d, was subjected to further mechanistic evaluation: it produced concentration-dependent inhibition zones in Oxford cup assays, exhibited bactericidal kinetics in time-kill studies, and significantly suppressed biofilm formation. Molecular docking suggested that the anti-biofilm activity of 3d may be mediated through binding to the staphylococcal accessory regulator A (SarA), a key transcriptional regulator of biofilm formation. The molecular dynamics study provided additional confirmation of the effective binding between 3d and SarA. These results highlight compound 3d as a promising lead for the development of novel anti-biofilm agents targeting drug-resistant Gram-positive infections.<\/jats:p>","DOI":"10.3390\/microorganisms14030574","type":"journal-article","created":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T15:15:29Z","timestamp":1772550929000},"page":"574","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Design, Synthesis, and Anti-Biofilm Activity of C-28 Modified Betulinic Acid Derivatives Targeting SarA in Drug-Resistant Staphylococcus aureus"],"prefix":"10.3390","volume":"14","author":[{"given":"Dongshun","family":"Jia","sequence":"first","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"},{"name":"School of Pharmacy, China Medical University, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junchao","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"},{"name":"School of Pharmacy, China Medical University, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuejin","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6877-6388","authenticated-orcid":false,"given":"Peng","family":"Gao","sequence":"additional","affiliation":[{"name":"Faculty of Dentistry, The University of Hong Kong, Hong Kong 999077, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongyu","family":"Zhan","sequence":"additional","affiliation":[{"name":"Faculty of Dentistry, The University of Hong Kong, Hong Kong 999077, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zihan","family":"Dong","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Li","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1331-2164","authenticated-orcid":false,"given":"Fanhao","family":"Meng","sequence":"additional","affiliation":[{"name":"School of Pharmacy, China Medical University, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6696-0966","authenticated-orcid":false,"given":"Nan","family":"Cai","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dajun","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Shenyang Medical College, Shenyang 110034, China"},{"name":"School of Pharmacy, China Medical University, Shenyang 110034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"990","DOI":"10.1038\/nature06536","article-title":"Global trends in emerging infectious diseases","volume":"451","author":"Jones","year":"2008","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1001\/jama.2018.19283","article-title":"Evaluation and Management of Penicillin Allergy","volume":"321","author":"Shenoy","year":"2019","journal-title":"JAMA"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"He, Y., Song, X., Wan, H., and Zhao, X. (2025). AmpHGT: Expanding prediction of antimicrobial activity in peptides containing non-canonical amino acids using multi-view constrained heterogeneous graph transformer. BMC Biol., 23.","DOI":"10.1186\/s12915-025-02253-4"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Abdelwahab, M.A., Amer, W.H., Elsharawy, D., Elkolaly, R.M., Helal, R.A.E.F., El Malla, D.A., Elfeky, Y.G., Bedair, H.A., Amer, R.S., and Abd-Elmonsef, M.E. (2023). Phenotypic and Genotypic Characterization of Methicillin Resistance in Staphylococci Isolated from an Egyptian University Hospital. Pathogens, 12.","DOI":"10.3390\/pathogens12040556"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1016\/S0140-6736(24)01867-1","article-title":"Global burden of bacterial antimicrobial resistance 1990\u20132021: A systematic analysis with forecasts to 2050","volume":"404","author":"Naghavi","year":"2024","journal-title":"Lancet"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1038\/s41579-021-00649-x","article-title":"Antibiotic resistance in the environment","volume":"20","author":"Larsson","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/S1473-3099(18)30605-4","article-title":"Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: A population-level modelling analysis","volume":"19","author":"Cassini","year":"2019","journal-title":"Lancet Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/S0140-6736(21)02724-0","article-title":"Global burden of bacterial antimicrobial resistance in 2019: A systematic analysis","volume":"399","author":"Murray","year":"2022","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e02225-19","DOI":"10.1128\/AAC.02225-19","article-title":"Antibiotic Resistance and Epigenetics: More to It than Meets the Eye","volume":"64","author":"Ghosh","year":"2020","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e011341","DOI":"10.1136\/bmjgh-2022-011341","article-title":"Global and regional burden of attributable and associated bacterial antimicrobial resistance avertable by vaccination: Modelling study","volume":"8","author":"Kim","year":"2023","journal-title":"BMJ Glob. Health"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.bcp.2016.11.002","article-title":"New approaches to antimicrobial discovery","volume":"134","author":"Lewis","year":"2017","journal-title":"Biochem. Pharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1038\/s41573-020-00114-z","article-title":"Natural products in drug discovery: Advances and opportunities","volume":"20","author":"Atanasov","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Yuan, Y., Lei, Y., Xu, M., Zhao, B., and Xu, S. (2025). Bioactive Terpenes from Marine Sponges and Their Associated Organisms. Mar. Drugs, 23.","DOI":"10.3390\/md23030096"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2235095","DOI":"10.1080\/14756366.2023.2235095","article-title":"Application of the Mannich reaction in the structural modification of natural products","volume":"38","author":"Pu","year":"2023","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Fu, J., Wang, Y., Sun, M., Xu, Y., and Chen, L. (2022). Antibacterial Activity and Components of the Methanol-Phase Extract from Rhizomes of Pharmacophagous Plant Alpinia officinarum Hance. Molecules, 27.","DOI":"10.3390\/molecules27134308"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Li, P., Huang, S., Xiao, S., Xu, Y., Wei, X., Xiao, J., Guo, Z., Yu, Q., and Liu, M. (2022). Antiviral Activities of Green Tea Components against Grouper Iridovirus Infection In Vitro and In Vivo. Viruses, 14.","DOI":"10.3390\/v14061227"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1186\/s12951-023-02006-x","article-title":"Anti-glioma effect of ginseng-derived exosomes-like nanoparticles by active blood\u2013brain-barrier penetration and tumor microenvironment modulation","volume":"21","author":"Kim","year":"2023","journal-title":"J. Nanobiotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.4155\/fmc-2018-0337","article-title":"Artemisinin and Its Derivatives; Ancient Tradition Inspiring the Latest Therapeutic Approaches Against Malaria","volume":"11","author":"Martino","year":"2019","journal-title":"Future Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Cho, A.Y.H., Chung, H., Romero-Parra, J., Kumar, P., Allar\u00e0, M., Ligresti, A., Gallardo-Garrido, C., Pessoa-Mahana, H., Fa\u00fandez, M., and Pessoa-Mahana, C.D. (2023). Motifs in Natural Products as Useful Scaffolds to Obtain Novel Benzo[d]imidazole-Based Cannabinoid Type 2 (CB2) Receptor Agonists. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241310918"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"107729","DOI":"10.1016\/j.biotechadv.2021.107729","article-title":"Plant synthetic biology for producing potent phyto-antimicrobials to combat antimicrobial resistance","volume":"48","author":"Tiwari","year":"2021","journal-title":"Biotechnol. Adv."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.1038\/nbt1206-1504","article-title":"Prospects for plant-derived antibacterials","volume":"24","author":"Lewis","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Wimmer, Z. (2025). Selected Pentacyclic Triterpenoids and Their Derivatives as Biologically Active Compounds. Molecules, 30.","DOI":"10.3390\/molecules30153106"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1080\/1028415X.2023.2180865","article-title":"Neuroprotective properties of Betulin, Betulinic acid, and Ursolic acid as triterpenoids derivatives: A comprehensive review of mechanistic studies","volume":"27","author":"Farzan","year":"2024","journal-title":"Nutr. Neurosci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1016\/j.jep.2008.09.001","article-title":"Ursolic, oleanolic and betulinic acids: Antibacterial spectra and selectivity indexes","volume":"120","author":"Fontanay","year":"2008","journal-title":"J. Ethnopharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"da Silva, G.N.S., Primon-Barros, M., Macedo, A.J., and Gnoatto, S.C.B. (2019). Triterpene Derivatives as Relevant Scaffold for New Antibiofilm Drugs. Biomolecules, 9.","DOI":"10.3390\/biom9020058"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108530","DOI":"10.1016\/j.steroids.2019.108530","article-title":"Antimicrobial properties of amine- and guanidine-functionalized derivatives of betulinic, ursolic and oleanolic acids: Synthesis and structure\/activity evaluation","volume":"154","author":"Spivak","year":"2020","journal-title":"Steroids"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Lombrea, A., Semenescu, A.-D., Magyari-Pavel, I.Z., Turks, M., Lugi\u0146ina, J., Peipi\u0146\u0161, U., Muntean, D., Dehelean, C.A., Dinu, S., and Danciu, C. (2023). Comparison of In Vitro Antimelanoma and Antimicrobial Activity of 2,3-Indolo-betulinic Acid and Its Glycine Conjugates. Plants, 12.","DOI":"10.3390\/plants12061253"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, C.M., Jhan, Y.L., Tsai, S.J., and Chou, C.H. (2016). The Pleiotropic Antibacterial Mechanisms of Ursolic Acid against Methicillin-Resistant Staphylococcus aureus (MRSA). Molecules, 21.","DOI":"10.3390\/molecules21070884"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"152933","DOI":"10.1016\/j.phymed.2019.152933","article-title":"Novel targets of pentacyclic triterpenoids in Staphylococcus aureus: A systematic review","volume":"73","author":"Chung","year":"2020","journal-title":"Phytomedicine"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e00272","DOI":"10.1002\/cbdv.202500272","article-title":"Synthesis and Evaluation of the Antibacterial Activity of N-Substituted Piperazine Flavonol Derivatives","volume":"22","author":"Wang","year":"2025","journal-title":"Chem. Biodivers."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"100307","DOI":"10.1016\/j.bioflm.2025.100307","article-title":"10058-F4 Mediated inhibition of the biofilm formation in multidrug-resistant Staphylococcus aureus","volume":"10","author":"Dodia","year":"2025","journal-title":"Biofilm"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e202301275","DOI":"10.1002\/cbdv.202301275","article-title":"Synthesis of hydrazide derivative of betulinic acid, its organometallic complexes, characterization and bioassay","volume":"21","author":"Batool","year":"2024","journal-title":"Chem. Biodivers."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"286","DOI":"10.2174\/1871520619666191014153554","article-title":"Synthesis, anticancer, and antibacterial activity of betulinic and betulonic acid C-28-triphenylphosphonium conjugates with variable alkyl linker length","volume":"20","author":"Tsepaeva","year":"2020","journal-title":"Anti-Cancer Agents Med. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1055\/s-1999-14054","article-title":"New triterpenoids with antibacterial activity from Zizyphus joazeiro","volume":"65","author":"Heilmann","year":"1999","journal-title":"Planta Med."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/3\/574\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T05:25:31Z","timestamp":1772688331000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/3\/574"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,3]]},"references-count":34,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["microorganisms14030574"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14030574","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,3]]}}}