{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T14:39:13Z","timestamp":1790606353905,"version":"4.1.0"},"reference-count":36,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T00:00:00Z","timestamp":1763683200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ANID Chile","award":["1220831"],"award-info":[{"award-number":["1220831"]}]},{"name":"ANID Chile","award":["11250944"],"award-info":[{"award-number":["11250944"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antioxidants"],"abstract":"<jats:p>Skin cancer is increasing worldwide, with melanoma being its most aggressive and lethal form due to its high metastatic potential. Despite therapeutic advances, drug resistance remains a challenge, highlighting the need to explore new anticancer agents. Leptocarpha rivularis is a native plant of Chile, locally called \u201cPalo negro\u201d, and is traditionally used in medicine by the Mapuche people. L. rivularis produces bioactive germacrene sesquiterpenoids with cytotoxic, antioxidant, anti-inflammatory and anti-angiogenic properties. This study reports for the first time the isolation of ovatifolin from aerial parts of L. rivularis and its identification by NMR and X-ray diffraction, together with its antiproliferative activity against two melanoma cell lines. The results show that ovatifolin has cytotoxic activity against the cell lines A2058 and A375, with an IC50 of 27.6 (90.2 \u00b5M) and 18.4 \u00b5g\/mL (60.1 \u00b5M), respectively, evaluated by live-cell IncuCyte\u00ae analysis. Moreover, ovatifolin arrests colony formation in a clonogenic assay, with an IC50 of 3.26 (10.6 \u03bcM) and 3.65 \u00b5g\/mL (11.9 \u03bcM) in these same cell lines. Therefore, ovatifolin increased intracellular ROS and decreased the mitochondrial membrane potential (\u0394\u03a8 m). Cell death studies using Annexin V showed that its cytotoxic activity is partially caused by non-specific apoptosis, which was corroborated by the caspase inhibitor Z-VAD with an incomplete recovery of the cell death process.<\/jats:p>","DOI":"10.3390\/antiox14121392","type":"journal-article","created":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T15:33:06Z","timestamp":1763739186000},"page":"1392","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Ovatifolin Purified from Leptocarpha rivularis Induces Cell Death in A375 and A2058 Melanoma Cancer Cells"],"prefix":"10.3390","volume":"14","author":[{"given":"Viviana","family":"Burgos","sequence":"first","affiliation":[{"name":"Escuela de Tecnolog\u00eda M\u00e9dica, Facultad de Salud, Universidad Santo Tom\u00e1s, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Cortez","sequence":"additional","affiliation":[{"name":"Laboratory of Natural Products & Drug Discovery, Center CEBIM, Department of Basic Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roc\u00edo","family":"Aguilera-Paill\u00e1n","sequence":"additional","affiliation":[{"name":"Laboratory of Natural Products & Drug Discovery, Center CEBIM, Department of Basic Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sof\u00eda","family":"Bravo-Bouchat","sequence":"additional","affiliation":[{"name":"Laboratory of Natural Products & Drug Discovery, Center CEBIM, Department of Basic Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0224-6069","authenticated-orcid":false,"given":"Bernd","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Chemie, Universit\u00e4t Potsdam, Karl-Liebknecht-Str. 24\u201325, D-14476 Potsdam, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3118-9518","authenticated-orcid":false,"given":"Eric","family":"Sperlich","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Chemie, Universit\u00e4t Potsdam, Karl-Liebknecht-Str. 24\u201325, D-14476 Potsdam, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebeca","family":"P\u00e9rez","sequence":"additional","affiliation":[{"name":"Carrera de Qu\u00edmica y Farmacia, Facultad de Ciencias de la Salud, Universidad Aut\u00f3noma de Chile, Avenida Alemania 01090, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nelia M.","family":"Rodriguez","sequence":"additional","affiliation":[{"name":"Carrera de Qu\u00edmica y Farmacia, Facultad de Ciencias de la Salud, Universidad Aut\u00f3noma de Chile, Avenida Alemania 01090, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8521-1469","authenticated-orcid":false,"given":"Leandro","family":"Ortiz","sequence":"additional","affiliation":[{"name":"Instituto de Ciencias Qu\u00edmicas, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5110566, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaime R.","family":"Cabrera-Pardo","sequence":"additional","affiliation":[{"name":"Laboratorio de Qu\u00edmica Aplicada y Sustentable (LabQAS), Departamento de Qu\u00edmica, Universidad del B\u00edo-B\u00edo, Avenida Collao 1202, Concepcion 4051381, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9066-1763","authenticated-orcid":false,"given":"Cecilia","family":"Villegas","sequence":"additional","affiliation":[{"name":"Departamento de Ciencias Biol\u00f3gicas y Qu\u00edmicas, Facultad de Recursos Naturales, Universidad Cat\u00f3lica de Temuco, Rudecindo Ortega, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4668-4984","authenticated-orcid":false,"given":"Cristian","family":"Paz","sequence":"additional","affiliation":[{"name":"Laboratory of Natural Products & Drug Discovery, Center CEBIM, Department of Basic Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco 4780000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,21]]},"reference":[{"key":"ref_1","first-page":"62","article-title":"Anatomy and physiology of the skin","volume":"17","author":"Nicol","year":"2005","journal-title":"Dermatol. Nurs.\/Dermatol. Nurses\u2019 Assoc."},{"key":"ref_2","unstructured":"Cassidy, P. (2016). Melanoma, Springer."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/j.jaad.2021.03.109","article-title":"United States Burden of Melanoma and Non-Melanoma Skin Cancer from 1990 to 2019","volume":"85","author":"Aggarwal","year":"2021","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Saginala, K., Barsouk, A., Aluru, J.S., Rawla, P., and Barsouk, A. (2021). Epidemiology of Melanoma. Med. Sci., 9.","DOI":"10.3390\/medsci9040063"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2118","DOI":"10.1002\/cncr.30435","article-title":"Mechanisms and Strategies to Overcome Resistance to Molecularly Targeted Therapy for Melanoma","volume":"123","author":"Lim","year":"2017","journal-title":"Cancer"},{"key":"ref_6","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_7","doi-asserted-by":"crossref","first-page":"184","DOI":"10.3390\/ddc3010011","article-title":"Plant-Derived Natural Products: A Source for Drug Discovery and Development","volume":"3","author":"Chaachouay","year":"2024","journal-title":"Drugs Drug Candidates"},{"key":"ref_8","first-page":"100460","article-title":"Three-Stage Sequential Supercritical Extraction in Fixed Bed Extractor to Obtain Bioactive Compounds from Leptocarpha rivularis Leaves","volume":"34","author":"Uquiche","year":"2023","journal-title":"J. Appl. Res. Med. Aromat. Plants"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Sheoran, S. (2024). Antioxidant Activity of Leptocarpha rivularis DC Flower Extract and Globularia alypum L. Authorea, 1\u201312.","DOI":"10.22541\/au.170665097.74741238\/v2"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"106314","DOI":"10.1016\/j.supflu.2024.106314","article-title":"Effect of Process Parameters on the Extraction Kinetics of Leptocarpha rivularis DC. in a Packed Bed Extractor Using Supercritical Carbon Dioxide","volume":"211","author":"Uquiche","year":"2024","journal-title":"J. Supercrit. Fluids"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Rubio, J., Arias, G., Robles-Kelly, C., Silva-Moreno, E., Espinoza, L., Carrasco, H., and Olea, A.F. (2022). Phytochemical Profiling and Assessment of Anticancer Activity of Leptocarpha rivularis Extracts Obtained from In Vitro Cultures. Plants, 11.","DOI":"10.3390\/plants11040546"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Rocha, \u00c1., Carlos, P., Juan, L.-L., and Salgado-Guerrero, P. (2021). Communication, Smart Technologies and Innovation for Society Proceedings of CITIS 2021, Springer.","DOI":"10.1007\/978-981-16-4126-8"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Carrasco, N., Garrido, M., Montenegro, I., Madrid, A., Hartley, R., Gonz\u00e1lez, I., Rubilar, M., Villena, J., and Valenzuela-Valderrama, M. (2023). Antitumoral Activity of Leptocarpha rivularis Flower Extracts against Gastric Cancer Cells. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021439"},{"key":"ref_14","unstructured":"STOE & CIE GMBH (2015). X-Area, version 1.75, STOE & CIE GMBH. Software Package for Collecting Single-Crystal Data on STOE Area-Detector Diffractometers, for Image Processing, Scaling Reflection Intensities and for Outlier Rejection."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1107\/S0108767307043930","article-title":"A short history of SHELX","volume":"64","author":"Sheldrick","year":"2008","journal-title":"Acta Crystallogr. Sect. A Found. Adv."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1107\/S1600576719014092","article-title":"Mercury 4.0: From visualization to analysis, design and prediction","volume":"53","author":"Macrae","year":"2020","journal-title":"Appl. Crystallogr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"110995","DOI":"10.1016\/j.celrep.2022.110995","article-title":"Oxidative Stress from DGAT1 Oncoprotein Inhibition in Melanoma Suppresses Tumor Growth When ROS Defenses Are Also Breached","volume":"39","author":"Wilcock","year":"2022","journal-title":"Cell Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Choudhry, P. (2016). High-Throughput Method for Automated Colony and Cell Counting by Digital Image Analysis Based on Edge Detection. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0148469"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2315","DOI":"10.1038\/nprot.2006.339","article-title":"Clonogenic Assay of Cells in Vitro","volume":"1","author":"Franken","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_20","first-page":"696","article-title":"Isolation and Characterization of Sesquiterpene Lactones from Calea Uniflora Less. And Their Leishmanicidal and Trypanocidal Activities","volume":"44","author":"Limaa","year":"2021","journal-title":"Qu\u00edmica Nova"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1328","DOI":"10.37349\/etat.2023.00200","article-title":"Cola rostrata K. Schum. Constituents Induce Cytotoxicity through Reactive Oxygen Species Generation\u00e3nd Mitochondrial Membrane Depolarisation","volume":"4","author":"Ajayi","year":"2023","journal-title":"Explor. Target. Anti-Tumor Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2469","DOI":"10.1016\/0031-9422(73)80456-X","article-title":"Sesquiterpene Lactones from Podanthus Ovatifolius","volume":"12","author":"Gnecco","year":"1973","journal-title":"Phytochemistry"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/BF01239677","article-title":"Ovatifolin, a Sesquiterpene Lactone","volume":"7","author":"Gopalakrishna","year":"1977","journal-title":"J. Cryst. Mol. Struct."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2765","DOI":"10.1016\/S0031-9422(00)83964-3","article-title":"Sesquiterpene Lactones of Podanthus Mitiqui","volume":"19","author":"Hoeneisen","year":"1980","journal-title":"Phytochemistry"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4243","DOI":"10.1021\/jf010351c","article-title":"Comparative Study of Ovatifolin Antioxidant and Growth Inhibition Activities","volume":"49","author":"Cespedes","year":"2001","journal-title":"J. Agric. Food Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1055\/s-2007-969474","article-title":"New Melampolides from Inula Germanica","volume":"51","author":"Bohlmann","year":"1985","journal-title":"Planta Medica"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.2298\/JSC210730087T","article-title":"Terpenoids in Four Inula Species from Bulgaria","volume":"86","author":"Trendafilova","year":"2021","journal-title":"J. Serbian Chem. Soc."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.1016\/S0031-9422(00)81746-X","article-title":"Germacranolides, Guaianolides and Eudesmanolides from Greenmaniella Resinosa","volume":"26","author":"Scott","year":"1987","journal-title":"Phytochemistry"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.1016\/0031-9422(95)00807-1","article-title":"Further Sesquiterpene Lactones from Inula Salsoloides","volume":"41","author":"Jeske","year":"1996","journal-title":"Phytochemistry"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1080\/14786419.2017.1329731","article-title":"A New Sesquiterpene Lactone Glucoside and Other Constituents from Inula salsoloides with Insecticidal Activities on Striped Flea Beetle (Phyllotreta striolata Fabricius)","volume":"32","author":"Bai","year":"2018","journal-title":"Nat. Prod. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.canlet.2006.03.004","article-title":"Isocostunolide, a Sesquiterpene Lactone, Induces Mitochondrial Membrane Depolarization and Caspase-Dependent Apoptosis in Human Melanoma Cells","volume":"246","author":"Chen","year":"2007","journal-title":"Cancer Lett."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/j.bcp.2007.08.024","article-title":"Induction of G2\/M Arrest and Apoptosis by Sesquiterpene Lactones in Human Melanoma Cell Lines","volume":"75","author":"Rozenblat","year":"2008","journal-title":"Biochem. Pharmacol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1006\/abio.2001.5116","article-title":"Fluorescence Microplate-Based Assay for Tumor Necrosis Factor Activity Using SYTOX Green Stain","volume":"293","author":"Jones","year":"2001","journal-title":"Anal. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/BF01702357","article-title":"Genotoxic Effects of Erioflorin Acetate and Erioflorin Methacrylate: Sesquiterpene Lactones Isolated from Podanthus Ovatifolius Lag. (Compositae)","volume":"44","author":"Cea","year":"1990","journal-title":"Bull. Environ. Contam. Toxicol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Villegas, C., Gonz\u00e1lez-Chavarr\u00eda, I., Burgos, V., Cabrera-Pardo, J.R., Schmidt, B., and Paz, C. (2025). Erioflorin and Erioflorin Acetate Induce Cell Death in Advanced Prostate Cancer Through ROS Increase and NF-\u039aB Inhibition. J. Xenobiot., 15.","DOI":"10.3390\/jox15020045"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"174989","DOI":"10.1016\/j.ejphar.2022.174989","article-title":"SCP-7, a Germacrane-Type Sesquiterpene Lactone Derivative, Induces ROS-Mediated Apoptosis in NSCLC Cells in Vitro and in Vivo","volume":"925","author":"Zhang","year":"2022","journal-title":"Eur. J. Pharmacol."}],"container-title":["Antioxidants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3921\/14\/12\/1392\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T10:35:17Z","timestamp":1764153317000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3921\/14\/12\/1392"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,21]]},"references-count":36,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["antiox14121392"],"URL":"https:\/\/doi.org\/10.3390\/antiox14121392","relation":{},"ISSN":["2076-3921"],"issn-type":[{"value":"2076-3921","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,21]]}}}