{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T16:16:28Z","timestamp":1791821788606,"version":"4.3.4"},"reference-count":42,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T00:00:00Z","timestamp":1619654400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003407","name":"Ministero dell\u2019Istruzione, dell\u2019Universit\u00e0 e della Ricerca","doi-asserted-by":"publisher","award":["FAR 2019"],"award-info":[{"award-number":["FAR 2019"]}],"id":[{"id":"10.13039\/501100003407","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Carvacrol (CAR), a phenolic monoterpenoid, has been extensively investigated for its antimicrobial and antifungal activity. As a result of its poor physicochemical properties, water soluble carvacrol prodrugs (WSCPs) with improved water solubility were previously synthesized and found to possess antimicrobial activity. Here, three novel CAR analogs, WSCP1, WSCP2, and WSCP3, were tested against fluconazole (FLU)-sensitive and -resistant strains where they showed greater antifungal activity than CAR against C. albicans. The probable mechanism by which the CAR prodrugs exert the antifungal activity was studied. Results from medium acidification assays demonstrated that the CAR and its synthetically designed prodrugs inhibit the yeast plasma membrane H+-ATPase (Pma1p), an essential target in fungi. In other words, in vitro data indicated that CAR analogs can prove to be a better alternative to CAR considering their improved water solubility. In addition, CAR and WSCP1 were developed into intravaginal formulations and administered at test doses of 50 mg\/kg in a mouse model of vulvovaginal candidiasis (VVC). Whereas the CAR and WSCP1 formulations both exhibited antifungal efficacy in the mouse model of VVC, the WSCP1 formulation was superior to CAR, showing a remarkable decrease in infection by ~120-fold compared to the control (infected, untreated animals). Taken together, a synthetically designed prodrug of CAR, namely WSCP1, proved to be a possible solution for poorly water-soluble drugs, an inhibitor of an essential yeast pump in vitro and an effective and promising antifungal agent in vivo.<\/jats:p>","DOI":"10.3390\/pharmaceutics13050633","type":"journal-article","created":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T04:30:11Z","timestamp":1619670611000},"page":"633","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Antifungal Activity of Novel Formulations Based on Terpenoid Prodrugs against C. albicans in a Mouse Model"],"prefix":"10.3390","volume":"13","author":[{"given":"Suvidha","family":"Menon","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John\u2019s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuyi","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John\u2019s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richa","family":"Vartak","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John\u2019s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ketankumar","family":"Patel","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John\u2019s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3042-2234","authenticated-orcid":false,"given":"Antonio","family":"Di Stefano","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, \u201cG. D\u2019Annunzio\u201d University of Chieti-Pescara, 66100 Chieti Scalo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6253-0443","authenticated-orcid":false,"given":"Ivana","family":"Cacciatore","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, \u201cG. D\u2019Annunzio\u201d University of Chieti-Pescara, 66100 Chieti Scalo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Marinelli","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, \u201cG. D\u2019Annunzio\u201d University of Chieti-Pescara, 66100 Chieti Scalo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7180-5955","authenticated-orcid":false,"given":"Blase","family":"Billack","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John\u2019s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.mib.2016.08.006","article-title":"Cell biology of Candida albicans-host interactions","volume":"34","author":"Lee","year":"2016","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1111\/1471-0528.12994","article-title":"Vulvovaginal Candida albicans infections: Pathogenesis, immunity and vaccine prospects","volume":"122","author":"Cassone","year":"2015","journal-title":"BJOG"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1016\/j.fct.2007.09.106","article-title":"Biological effects of essential oils\u2014A review","volume":"46","author":"Bakkali","year":"2008","journal-title":"Food Chem. Toxicol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"113318","DOI":"10.1016\/j.jep.2020.113318","article-title":"Traditional use, phytochemistry, toxicology, and pharmacology of Origanum majorana L.","volume":"265","author":"Bouyahya","year":"2021","journal-title":"J. Ethnopharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.3855\/jidc.12469","article-title":"Biological activities of essential oil extracted from leaves of Atalantia sessiflora Guillauminin Vietnam","volume":"14","author":"Le","year":"2020","journal-title":"J. Infect. Dev. Ctries."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2016\/3012462","article-title":"Antimicrobial properties of plant essential oils against human pathogens and their mode of action: An updated review","volume":"6","author":"Swamy","year":"2016","journal-title":"Evid. Based Complement. Alternat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1046\/j.1365-2672.2001.01428.x","article-title":"A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol","volume":"91","author":"Lambert","year":"2001","journal-title":"J. Appl. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3989","DOI":"10.3390\/molecules17043989","article-title":"Essential Oils in Combination and Their Antimicrobial Properties","volume":"17","author":"Juliani","year":"2012","journal-title":"Molecules"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Cacciatore, I., di Giulio, M., Fornasari, E., di Stefano, A., Cerasa, L.S., Marinelli, L., Turkez, H., di Campli, E., di Bartolomeo, S., and Robuffo, I. (2015). Carvacrol codrugs: A new approach in the antimicrobial plan. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0120937"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"7652","DOI":"10.1021\/jf5023862","article-title":"Chemistry and Multibeneficial Bioactivities of Carvacrol (4-Isopropyl-2-methylphenol), a Component of Essential Oils Produced by Aromatic Plants and Spices","volume":"62","author":"Friedman","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1874","DOI":"10.1021\/jf052564y","article-title":"Structural Requirements for the Antimicrobial Activity of Carvacrol","volume":"54","author":"Veldhuizen","year":"2006","journal-title":"J. Agric. Food Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1128\/AEM.68.4.1561-1568.2002","article-title":"The Phenolic Hydroxyl Group of Carvacrol Is Essential for Action against the Food-Borne Pathogen Bacillus cereus","volume":"68","author":"Ultee","year":"2002","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1007\/s10096-010-1050-8","article-title":"Fungicidal activity of thymol and carvacrol by disrupting ergosterol biosynthesis and membrane integrity against Candida","volume":"30","author":"Ahmad","year":"2011","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.ejmech.2019.05.093","article-title":"Carvacrol prodrugs as novel antimicrobial agents","volume":"178","author":"Marinelli","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1111\/j.1749-6632.1997.tb52330.x","article-title":"The plasma membrane H(+)-ATPase of fungi. A candidate drug target?","volume":"834","author":"Perlin","year":"1997","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00684-17","DOI":"10.1128\/IAI.00684-17","article-title":"Novel mechanism behind the immunopathogenesis of vulvovaginal candidiasis: \u201cNeutrophil anergy\u201d","volume":"86","author":"Yano","year":"2018","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"19.6.1","DOI":"10.1002\/0471142735.im1906s105","article-title":"Animal Models for Candidiasis","volume":"105","author":"Conti","year":"2014","journal-title":"Curr. Protoc. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1046\/j.1439-0507.1999.00498.x","article-title":"Molecular aspects of fluconazole resistance development in Candida albicans","volume":"42","author":"Franz","year":"1999","journal-title":"Mycoses"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1590\/S0036-46652008000100002","article-title":"Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates","volume":"50","author":"Lyon","year":"2008","journal-title":"Rev. Inst. Med. Trop. South Paulo."},{"key":"ref_20","unstructured":"Clinical and Laboratory Standards Institute (2002). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts. Approved Standard, Clinicaland Laboratory Standards Institute. [2nd ed.]. NCCLS Document M27-A2."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"18118","DOI":"10.1016\/S0021-9258(19)81331-5","article-title":"Membrane potential defect in hygromycin B-resistant pma1 mutants of Saccharomyces cerevisiae","volume":"263","author":"Perlin","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e21892","DOI":"10.1002\/jbt.21892","article-title":"In vitro assessment of the growth and plasma membrane H + -ATPase inhibitory activity of ebselen and structurally related selenium- and sulfur-containing compounds in Candida albicans","volume":"31","author":"Orie","year":"2017","journal-title":"J. Biochem. Mol. Toxicol."},{"key":"ref_23","first-page":"3382","article-title":"Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis","volume":"58","author":"Yano","year":"2011","journal-title":"J. Vis. Exp."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1080\/01480545.2018.1488858","article-title":"Ebselen oxide attenuates mechlorethamine dermatotoxicity in the mouse ear vesicant model","volume":"43","author":"Tumu","year":"2020","journal-title":"Drug Chem. Toxicol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1002\/jobm.201100272","article-title":"Proton pumping ATPase mediated fungicidal activity of two essential oil components","volume":"52","author":"Bhatia","year":"2012","journal-title":"J. Basic Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0304-4157(88)90017-2","article-title":"Structure and function of proton translocating ATPase in plasma membranes of plants and fungi","volume":"947","author":"Serrano","year":"1988","journal-title":"Biochim. Biophys. Acta."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1016\/j.fitote.2010.07.020","article-title":"Proton translocating ATPase mediated fungicidal activity of eugenol and thymol","volume":"81","author":"Ahmad","year":"2010","journal-title":"Fitoterapia"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"105323","DOI":"10.1016\/j.ejps.2020.105323","article-title":"Ebselen nanoemulgel for the treatment of topical fungal infection","volume":"148","author":"Vartak","year":"2020","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2835","DOI":"10.2217\/nnm-2019-0243","article-title":"A vaginal nanoformulation of a SphK inhibitor attenuates lipopolysaccharide-induced preterm birth in mice","volume":"14","author":"Giusto","year":"2019","journal-title":"Nanomedicine"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1128\/IAI.68.2.651-657.2000","article-title":"Effects of Reproductive Hormones on Experimental Vaginal Candidiasis","volume":"68","author":"Fidel","year":"2000","journal-title":"Infect. Immun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1139\/W08-097","article-title":"In vitro activity of essential oils extracted from plants used as spices against fluconazole-resistant and fluconazole-susceptible Candida spp.","volume":"54","author":"Pozzatti","year":"2008","journal-title":"Can. J. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.1111\/jam.13282","article-title":"Sensitivity of Candida albicans to essential oils: Are they an alternative to antifungal agents?","volume":"121","author":"Bona","year":"2016","journal-title":"J. Appl. Microbiol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Nazzaro, F., Fratianni, F., Coppola, R., and Feo, V. (2017). Essential oils and antifungal Activity. Pharmaceuticals, 10.","DOI":"10.3390\/ph10040086"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.vetpar.2016.01.001","article-title":"Comparative efficacy and toxic effects of carvacryl acetate and carvacrol on sheep gastrointestinal nematodes and mice","volume":"218","author":"Andre","year":"2016","journal-title":"Vet. Parasitol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4079425","DOI":"10.1155\/2017\/4079425","article-title":"The neuroprotective effects of carvacrol on ethanol-induced hippocampal neurons impairment via the antioxidative and antiapoptotic pathways","volume":"2017","author":"Wang","year":"2017","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1722","DOI":"10.1111\/j.2042-7158.2012.01552.x","article-title":"Antinociceptive activity of carvacrol (5-isopropyl-2-methylphenol) in mice","volume":"64","author":"Rios","year":"2012","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1128\/EC.00103-08","article-title":"A gain-of-function mutation in the transcription factor Upc2p causes upregulation of ergosterol biosynthesis genes and increased fluconazole resistance in a clinical Candida albicans isolate","volume":"7","author":"Dunkel","year":"2008","journal-title":"Eukaryot. Cell."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2676","DOI":"10.1128\/AAC.45.10.2676-2684.2001","article-title":"Prevalence of molecular mechanisms of resistance to azole antifungal agents in Candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients","volume":"45","author":"Perea","year":"2001","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_39","first-page":"401","article-title":"Inhibitory effects of carvacrol on the expression of secreted aspartyl proteinases 1-3 in fluconazole-resistant Candida albicans isolates","volume":"8","author":"Hosseini","year":"2016","journal-title":"Iran. J. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5062","DOI":"10.1128\/AAC.01050-10","article-title":"Mechanism of antifungal activity of terpenoid phenols resembles calcium stress and inhibition of the TOR pathway","volume":"54","author":"Rao","year":"2010","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1111\/j.1472-765X.2008.02407.x","article-title":"The antibacterial mechanism of carvacrol and thymol against Escherichia coli","volume":"47","author":"Xu","year":"2008","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e00147-20","DOI":"10.1128\/JVI.00147-20","article-title":"Oregano Oil and Its Principal Component, Carvacrol, Inhibit HIV-1 Fusion into Target Cells","volume":"94","author":"Mediouni","year":"2020","journal-title":"J. Virol."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/13\/5\/633\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:55:06Z","timestamp":1760162106000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/13\/5\/633"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,29]]},"references-count":42,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["pharmaceutics13050633"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics13050633","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,29]]}}}