{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T20:26:40Z","timestamp":1783369600315,"version":"3.54.6"},"reference-count":30,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T00:00:00Z","timestamp":1745539200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Ocular fungal infections are slow-progressing conditions that primarily affect the cornea but can also involve the entire eyeball. Candida albicans is one of the most involved species. Both diagnosing and treating these infections require prompt and effective action. However, the currently available treatment options mainly rely on azoles and polyenes, which are known for their poor penetration into ocular tissue and associated toxicity. Moreover, conventional antifungals are usually ineffective when tested against biofilm-associated infections, mainly due to the metabolically inactive state of dormant cells embedded in the extracellular biofilm matrix. Here, analysis of the in vitro antifungal activity of four 2-aminobenzoic acid derivatives synthesized using a green method and their combination with Fluconazole (FLC) showed efficacy against the FLC-resistant clinical isolate of C. albicans under both planktonic and biofilm formation conditions. Results showed that compounds 1 and 2 exhibited the best antifungal activity in the checkerboard association test, presenting a synergistic effect towards antifungal action. The downregulation of HWP, ERG11, and ASL3 genes during biofilm inhibition suggested a reduced capacity of the four compounds for hyphal growth and adhesion, as well as a decrease in pathogenicity due to the downregulation of some SAP genes. In vitro and in vivo toxicity profiles indicated that these compounds exhibited low toxicity, as well as the absence of genotoxic effects. Therefore, green-synthetized 2-aminobenzoic acid derivatives may have potential as antifungal agents for the inhibition of C. albicans growth and biofilm formation.<\/jats:p>","DOI":"10.3390\/antibiotics14050432","type":"journal-article","created":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T04:08:24Z","timestamp":1745554104000},"page":"432","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Antifungal and Antibiofilm Activities of 2-Aminobenzoic Acid Derivatives Against a Clinical Ocular Candida albicans Isolate for Biomedical Applications"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0446-2846","authenticated-orcid":false,"given":"Francesco","family":"Petrillo","sequence":"first","affiliation":[{"name":"Department of Medical Sciences, Eye Clinic, Turin University, 10024 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4981-8837","authenticated-orcid":false,"given":"Angela","family":"Maione","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Naples \u2018Federico II\u2019, Via Cinthia, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marisa","family":"Spampinato","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Naples \u2018Federico II\u2019, Via Cinthia, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea Di","family":"Massa","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Naples \u2018Federico II\u2019, Via Cinthia, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6805-0408","authenticated-orcid":false,"given":"Marco","family":"Guida","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Naples \u2018Federico II\u2019, Via Cinthia, 80126 Naples, Italy"},{"name":"BAT Center\u2014Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology, University of Naples Federico II, 80055 Portici, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7866-821X","authenticated-orcid":false,"given":"Armando","family":"Zarrelli","sequence":"additional","affiliation":[{"name":"Department of Chemical Science, University of Napoli Federico II, Via Cinthia 4, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilia","family":"Galdiero","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Naples \u2018Federico II\u2019, Via Cinthia, 80126 Naples, Italy"},{"name":"BAT Center\u2014Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology, University of Naples Federico II, 80055 Portici, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5910-6358","authenticated-orcid":false,"given":"Luigi","family":"Longobardo","sequence":"additional","affiliation":[{"name":"Department of Chemical Science, University of Napoli Federico II, Via Cinthia 4, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,25]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2022). WHO Fungal Priority Pathogens List to Guide Research, Development and Public Health Action, WHO."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"759","DOI":"10.2174\/1568026618666180528121707","article-title":"Emerging Virulence, Drug Resistance and Future Anti-fungal Drugs for Candida Pathogens","volume":"18","author":"Vartika","year":"2018","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1038\/s41579-022-00767-0","article-title":"The biofilm life cycle: Expanding the conceptual model of biofilm formation","volume":"20","author":"Sauer","year":"2022","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mib.2019.04.001","article-title":"Candida albicans biofilm growth and dispersal: Contributions to pathogenesis","volume":"52","author":"Wall","year":"2019","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1126\/science.aap7999","article-title":"Worldwide emergence of resistance to antifungal drugs challenges human health and food security","volume":"360","author":"Fisher","year":"2018","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1021\/acs.jnatprod.9b01285","article-title":"Natural Products as Sources of New Drugs over the Nearly Four Decades from 01\/1981 to 09\/2019","volume":"83","author":"Newman","year":"2020","journal-title":"J. Nat. Prod."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Souza, L., de Oliveira Bento, A., Louren\u00e7o, E.M.G., Ferreira, M.R.A., Oliveira, W.N., Soares, L.A.L., Barbosa, E.G., Rocha, H.A.O., and Chaves, G.M. (2024). Mechanism of action and synergistic effect of Eugenia uniflora extract in Candida spp.. PLoS ONE, 19.","DOI":"10.1371\/journal.pone.0303878"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1002\/ddr.21842","article-title":"Medicinal chemistry of anthranilic acid derivatives: A mini review","volume":"82","author":"Prasher","year":"2021","journal-title":"Drug Dev. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"S3804","DOI":"10.1016\/j.arabjc.2014.05.018","article-title":"Synthesis, antimicrobial evaluation and QSAR studies of p-coumaric acid derivatives","volume":"10","author":"Khatkar","year":"2017","journal-title":"Arab. J. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e202305046","DOI":"10.1002\/slct.202305046","article-title":"Sustainable Chemical Derivatization of Hydroxycinnamic Acids","volume":"9","author":"Longobardo","year":"2024","journal-title":"ChemistrySelect"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Maione, A., Imparato, M., Buonanno, A., Carraturo, F., Schettino, A., Schettino, M.T., Galdiero, M., de Alteriis, E., Guida, M., and Galdiero, E. (2023). Anti-Biofilm Activity of Phenyllactic Acid against Clinical Isolates of Fluconazole-Resistant Candida albicans. J. Fungi, 9.","DOI":"10.3390\/jof9030355"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Cavalheiro, M., and Teixeira, M.C. (2018). Candida Biofilms: Threats, Challenges, and Promising Strategies. Front. Med., 5.","DOI":"10.3389\/fmed.2018.00028"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1007\/s12281-021-00419-5","article-title":"Investigational Agents for the Treatment of Resistant Yeasts and Molds","volume":"15","author":"Seiler","year":"2021","journal-title":"Curr. Fungal Infect. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"749","DOI":"10.2174\/138955712801264792","article-title":"Antioxidant and antimicrobial activities of cinnamic acid derivatives","volume":"12","author":"Sova","year":"2012","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1007\/s00044-010-9540-x","article-title":"QSAR studies on a number of pyrrolidin-2-one antiarrhythmic arylpiperazinyls","volume":"21","author":"Nowaczyk","year":"2012","journal-title":"Med. Chem. Res."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"de Morais, M.C., de Oliveira Lima, E., Perez-Castillo, Y., and de Sousa, D.P. (2023). Synthetic Cinnamides and Cinnamates: Antimicrobial Activity, Mechanism of Action, and In Silico Study. Molecules, 28.","DOI":"10.3390\/molecules28041918"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Liu, Y., Ren, H., Wang, D., Zhang, M., Sun, S., and Zhao, Y. (2020). The synergistic antifungal effects of gypenosides combined with fluconazole against resistant Candida albicans via inhibiting the drug efflux and biofilm formation. Biomed. Pharmacother., 130.","DOI":"10.1016\/j.biopha.2020.110580"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Manoharan, R.K., Lee, J.-H., Kim, Y.-G., and Lee, J. (2017). Alizarin and Chrysazin Inhibit Biofilm and Hyphal Formation by Candida albicans. Front. Cell. Infect. Microbiol., 7.","DOI":"10.3389\/fcimb.2017.00447"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1111\/j.1365-2958.1994.tb00429.x","article-title":"Multiplicity of genes encoding secreted aspartic proteinases in Candida species","volume":"13","author":"Monod","year":"1994","journal-title":"Mol. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5215","DOI":"10.1128\/jb.177.18.5215-5221.1995","article-title":"Candida albicans secreted aspartyl proteinases: Isoenzyme pattern is determined by cell type, and levels are determined by environmental factors","volume":"177","author":"White","year":"1995","journal-title":"J. Bacteriol."},{"key":"ref_21","unstructured":"CLSI (2018). Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts, Approved Guideline, Clinical and Laboratory Standards Institute. CLSI Doc. M44-A."},{"key":"ref_22","unstructured":"CLSI (2017). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Clinical and Laboratory Standards Institute. [4th ed.]."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"S37","DOI":"10.1016\/S0190-9622(98)70482-7","article-title":"Fungicidal versus fungistatic activity of terbinafine and itraconazole: An in vitro comparison","volume":"38","author":"Hazen","year":"1998","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Falanga, A., Maione, A., La Pietra, A., de Alteriis, E., Vitale, S., Bellavita, R., Carotenuto, R., Turr\u00e0, D., Galdiero, S., and Galdiero, E. (2022). Competitiveness during Dual-Species Biofilm Formation of Fusarium oxysporum and Candida albicans and a Novel Treatment Strategy. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14061167"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Tan, J., Jiang, S., Tan, L., Shi, H., Yang, L., Sun, Y., and Wang, X. (2021). Antifungal Activity of Minocycline and Azoles Against Fluconazole-Resistant Candida Species. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.649026"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/S0167-7012(00)00122-6","article-title":"A modified microtiter-plate test for quantification of staphylococcal biofilm formation","volume":"40","author":"Stepanovic","year":"2000","journal-title":"J. Microbiol. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e36","DOI":"10.1093\/nar\/30.9.e36","article-title":"Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR","volume":"30","author":"Pfaffl","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Buonanno, A., Salvatore, M.M., Feola, A., Siciliano, A., Bellavita, R., Imb\u00f2, L.E., Guida, M., Andolfi, A., Nicoletti, R., and Maione, A. (2024). Sphaeropsidin A Loaded in Liposomes to Reduce Its Cytotoxicity and Preserve Antifungal Activity Against Candida auris. Molecules, 29.","DOI":"10.3390\/molecules29245949"},{"key":"ref_30","unstructured":"(2013). Soil Quality\u2014Assessment of Genotoxic Effects on Higher plants\u2014Vicia faba Micronucleus Test. Standard No. ISO 29200:2013."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/5\/432\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:21:15Z","timestamp":1760030475000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/5\/432"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,25]]},"references-count":30,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["antibiotics14050432"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics14050432","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,25]]}}}