{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:44:24Z","timestamp":1760060664006,"version":"build-2065373602"},"reference-count":58,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T00:00:00Z","timestamp":1757462400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NHMRC Investigator Fellowship","award":["1174145"],"award-info":[{"award-number":["1174145"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Tyrosinase catalyzes the rate-limiting steps of melanin production, posing as an important target for treating skin pigmentation. This study investigates bioactive human tyrosinase inhibitors from Xanthium strumarium L. using a combined strategy of cell lysate, cell-based, and zebrafish assays. In this study, the methanol extract of Xanthium strumarium L. was identified as a potent inhibitor against tyrosinase in a cell lysate assay utilizing human MM418C1 melanoma cells. Subsequent phytochemical analysis resulted in the isolation of 11 natural products, including 4-hydroxybenzoic acid (4HB), three nucleotides, four caffeoylquinic acids and three alkaloids. Biological activity evaluation of isolated compounds suggested that 4HB was a potent inhibitor against tyrosinase with an IC50 value of 59.5 \u03bcg\/mL. Further evaluations revealed that 4HB significantly reduced the melanin content by 40% at the concentration of 500 mg\/mL in human MM418C1 melanoma cells. 4HB activity was finally confirmed in vivo, by the demonstration of 40% reduction in melanin production in live zebrafish at the concentration of 15.63 \u03bcg\/mL.<\/jats:p>","DOI":"10.3390\/molecules30183689","type":"journal-article","created":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T14:14:55Z","timestamp":1757513695000},"page":"3689","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation and Biological Evaluation of Human Tyrosinase Inhibitors from the Fruit of Xanthium strumarium L."],"prefix":"10.3390","volume":"30","author":[{"given":"Gengxuan","family":"Shi","sequence":"first","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaoying","family":"Lu","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yougang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ke","family":"Zheng","sequence":"additional","affiliation":[{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jean","family":"Giacomotto","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"},{"name":"Queensland Brain Institute, The University of Queensland, Brisbane, QLD 4067, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1018-2798","authenticated-orcid":false,"given":"Kathryn F.","family":"Tonissen","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2412-1213","authenticated-orcid":false,"given":"Yunjiang","family":"Feng","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University, Nathan, QLD 4111, Australia"},{"name":"School of Environment and Science, Griffith University, Nathan, QLD 4111, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2638","DOI":"10.2174\/0929867329666220915123714","article-title":"Medicinal prospects of targeting tyrosinase: A feature review","volume":"30","author":"Wang","year":"2023","journal-title":"Curr. Med. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1002\/chem.201704410","article-title":"Structure and function of human tyrosinase and tyrosinase-related proteins","volume":"24","author":"Lai","year":"2018","journal-title":"Chem.\u2013A Eur. J."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Solano, F. (2018). On the metal cofactor in the tyrosinase family. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020633"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"468","DOI":"10.2340\/0001555563468475","article-title":"Dopa oxidation and tyrosine oxygenation by human melanoma tyrosinase","volume":"63","author":"Jergil","year":"1983","journal-title":"Acta Derm.-Venereol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Couteau, C., and Coiffard, L. (2016). Overview of skin whitening agents: Drugs and cosmetic products. Cosmetics, 3.","DOI":"10.3390\/cosmetics3030027"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1111\/j.1529-8019.2007.00144.x","article-title":"Skin lightening preparations and the hydroquinone controversy","volume":"20","author":"Draelos","year":"2007","journal-title":"Dermatol. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.7573\/dic.2021-11-2","article-title":"Dermatology: How to manage facial hyperpigmentation in skin of colour","volume":"11","author":"Moolla","year":"2022","journal-title":"Drugs Context"},{"key":"ref_8","unstructured":"Bernauer, U., Bodin, L., Chaudhry, Q., Coenraads, P.J., Dusinska, M., Ezendam, J., Gaffet, E., Galli, C.L., Panteri, E., and Rogiers, V. (2023). SCCS OPINION on the Safety of Alpha-Arbutin and Beta-Arbutin in Cosmetic Products-SCCS\/1642\/22\u2013Final Opinion, European Commission."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Ayuhastuti, A., Syah, I.S.K., Megantara, S., and Chaerunisaa, A.Y. (2023). A Comprehensive review of the cosmetic application of kojic acid dipalmitate: Kojic acid derivative with improved properties. arXiv.","DOI":"10.20944\/preprints202312.2196.v1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"13428","DOI":"10.1021\/acs.jmedchem.0c00994","article-title":"Advances in the design of genuine human tyrosinase inhibitors for targeting melanogenesis and related pigmentations","volume":"63","author":"Roulier","year":"2020","journal-title":"J. Med. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1080\/09546634.2020.1800579","article-title":"The versatility of azelaic acid in dermatology","volume":"33","author":"Searle","year":"2022","journal-title":"J. Dermatol. Treat."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1080\/07366205.2024.2441637","article-title":"Development of a human tyrosinase activity inhibition assay using human melanoma cell lysate","volume":"76","author":"Shi","year":"2024","journal-title":"BioTechniques"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1271\/bbb.110894","article-title":"Inhibition of melanogenesis by Xanthium strumarium L. Biosci","volume":"76","author":"Li","year":"2012","journal-title":"Biotechnol. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Iqbal, M.F., Liu, M.-C., Iram, A., Feng, Y.-L., and Xu, F. (2020). Effects of the invasive plant Xanthium strumarium on diversity of native plant species: A competitive analysis approach in North and Northeast China. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0228476"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Fan, W., Fan, L., Peng, C., Zhang, Q., Wang, L., Li, L., Wang, J., Zhang, D., Peng, W., and Wu, C. (2019). Traditional uses, botany, phytochemistry, pharmacology, pharmacokinetics and toxicology of Xanthium strumarium L.: A review. Molecules, 24.","DOI":"10.3390\/molecules24020359"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1526","DOI":"10.1021\/np500951s","article-title":"Bioactive sesquiterpenes and lignans from the fruits of Xanthium sibiricum","volume":"78","author":"Shi","year":"2015","journal-title":"J. Nat. Prod."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"814","DOI":"10.1007\/s10600-009-9208-2","article-title":"Composition of supercritical fluid extracts of some Xanthium species from China","volume":"44","author":"Han","year":"2008","journal-title":"Chem. Nat. Compd."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4746","DOI":"10.1016\/j.bmc.2015.05.044","article-title":"Sesquiterpenes with TRAIL-resistance overcoming activity from Xanthium strumarium","volume":"23","author":"Karmakar","year":"2015","journal-title":"Bioorg. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1080\/10286020.2015.1070146","article-title":"A new thiophene and two new monoterpenoids from Xanthium sibiricum","volume":"17","author":"Shi","year":"2015","journal-title":"J. Asian Nat. Prod. Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Hwang, S.H., Wang, Z., Na Yoon, H., and Lim, S.S. (2016). Xanthium strumarium as an inhibitor of \u03b1-glucosidase, protein tyrosine phosphatase 1\u03b2, protein glycation and ABTS+ for diabetic and its complication. Molecules, 21.","DOI":"10.3390\/molecules21091241"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1007\/s10600-006-0215-2","article-title":"New thiazinediones and other components from Xanthium strumarium","volume":"42","author":"Han","year":"2006","journal-title":"Chem. Nat. Compd."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.bcab.2018.02.004","article-title":"Optimization of extraction conditions and assessment of antioxidant, \u03b1-glucosidase inhibitory and antimicrobial activities of Xanthium strumarium L. fruits","volume":"14","author":"Ingawale","year":"2018","journal-title":"Biocatal. Agric. Biotechnol."},{"key":"ref_23","unstructured":"Sultana, A. (2014). Phytochemical Studies on the Chemical Constituents of Xanthium strumarium Linn., Synthesis in Addition Bioactivities of 2, 3-Diaminonaphthalenimidazole Derivatives and Amides of Piperic Acid. [Ph.D. Thesis, Federal Urdu University of Arts, Science and Technology]."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1080\/14786419.2010.539182","article-title":"Chemical constituents from the roots of Xanthium sibiricum","volume":"25","author":"Kan","year":"2011","journal-title":"Nat. Prod. Product Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1016\/0031-9422(93)85551-2","article-title":"1, 3, 5-Tri-O-caffeoylquinic acid from Xanthium strumarium","volume":"33","author":"Agata","year":"1993","journal-title":"Phytochemistry"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1080\/10286020.2015.1099525","article-title":"Two new compounds from Xanthium strumarium","volume":"18","author":"Yin","year":"2016","journal-title":"J. Asian Nat. Prod. Res."},{"key":"ref_27","first-page":"266","article-title":"Isolation and identification of phytochemicals from Xanthium strumarium","volume":"4","author":"Pandey","year":"2012","journal-title":"Int. J. Chem Tech Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"12464","DOI":"10.3390\/molecules181012464","article-title":"Four new glycosides from the fruit of Xanthium sibiricum Patr","volume":"18","author":"Jiang","year":"2013","journal-title":"Molecules"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1016\/0031-9422(76)85126-6","article-title":"Isolation and identification of the hypoglycemic agent, carboxyatracrylate, from Xanthium strumarium","volume":"15","author":"Craig","year":"1976","journal-title":"Phytochemistry"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"408","DOI":"10.4103\/wjtcm.wjtcm_49_20","article-title":"The fruits of Xanthium sibiricum Patr: A review on phytochemistry, pharmacological activities, and toxicity","volume":"6","author":"Wang","year":"2020","journal-title":"World J. Tradit. Chin. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.ijpharm.2010.09.035","article-title":"In vitro and in vivo antioxidant properties of chlorogenic acid and caffeic acid","volume":"403","author":"Sato","year":"2011","journal-title":"Int. J. Pharm."},{"key":"ref_32","first-page":"67","article-title":"Amelioration of inflammatory responses by chlorogenic acid via suppression of pro-inflammatory mediators","volume":"1","author":"Chauhan","year":"2011","journal-title":"J. Appl. Pharm. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Liang, N., and Kitts, D.D. (2015). Role of chlorogenic acids in controlling oxidative and inflammatory stress conditions. Nutrients, 8.","DOI":"10.3390\/nu8010016"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.jff.2017.10.047","article-title":"Antioxidant and anti-inflammatory activities of quercetin and its derivatives","volume":"40","author":"Lesjak","year":"2018","journal-title":"J. Funct. Foods"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Gligor, O., Clichici, S., Moldovan, R., Muntean, D., Vlase, A.-M., Nad\u0103\u0219, G.C., Filip, G.A., Vlase, L., and Cri\u0219an, G. (2022). Influences of different extraction techniques and their respective parameters on the phytochemical profile and biological activities of Xanthium spinosum L. extracts. Plants, 12.","DOI":"10.3390\/plants12010096"},{"key":"ref_36","first-page":"1717","article-title":"Chemical constituents from Xanthii fructus","volume":"44","author":"Chen","year":"2013","journal-title":"Chin. Tradit. Herb. Drugs"},{"key":"ref_37","first-page":"47","article-title":"Chemical constituents from fruits of Xanthium sibiricum","volume":"48","author":"Jiang","year":"2017","journal-title":"Chin. Tradit. Herb. Drugs"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1187","DOI":"10.1007\/s10854-023-10494-8","article-title":"Crystal growth, structural, DFT, optical and Z-scan analyses of 4-hydroxybenzoic acid-1H-imidazole crystal","volume":"34","author":"Chinnakannu","year":"2023","journal-title":"J. Mater. Sci. Mater. Electron."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1007\/s10953-006-9114-1","article-title":"Interference of copper (II) ions with non-covalent interactions in uridine or uridine 5\u2032-monophosphate systems with adenosine, cytidine, thymidine and their monophosphates in aqueous solution","volume":"36","author":"Lomozik","year":"2007","journal-title":"J. Solut. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"6998","DOI":"10.1021\/acs.jafc.0c02183","article-title":"Sustainable straightforward synthesis and evaluation of the antioxidant and antimicrobial activity of sinapine and analogues","volume":"68","author":"Mouterde","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"112432","DOI":"10.1016\/j.phytochem.2020.112432","article-title":"Isolation, structure elucidation and neuroprotective effects of caffeoylquinic acid derivatives from the roots of Arctium lappa L.","volume":"177","author":"Gao","year":"2020","journal-title":"Phytochemistry"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.molstruc.2013.01.063","article-title":"A new series of Schiff bases derived from sulfa drugs and indole-3-carboxaldehyde: Synthesis, characterization, spectral and DFT computational studies","volume":"1039","author":"Ebrahimi","year":"2013","journal-title":"J. Mol. Struct."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1111\/j.1600-0749.1998.tb00735.x","article-title":"Does \u03b1-MSH have a role in regulating skin pigmentation in humans?","volume":"11","author":"Thody","year":"1998","journal-title":"Pigment. Cell Res."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Chung, S., Lim, G.J., and Lee, J.Y. (2019). Quantitative analysis of melanin content in a three-dimensional melanoma cell culture. Sci. Rep., 9.","DOI":"10.1038\/s41598-018-37055-y"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Wang, W., Yang, L., Wang, W., Zhang, J., Engelhardt, U.H., and Jiang, H. (2023). Inhibitory activities of samples on tyrosinases were affected by enzyme species and sample addition methods. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24076013"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"410","DOI":"10.3390\/ddc3020025","article-title":"Cyclocurcumin, a minor curcuminoid, is a novel candidate for hypopigmentary skin disorders with melanogenesis-stimulating capacity","volume":"3","author":"Goenka","year":"2024","journal-title":"Drugs Drug Candidates"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.aca.2004.07.001","article-title":"Preparation and characterization of molecularly imprinted monolithic column based on 4-hydroxybenzoic acid for the molecular recognition in capillary electrochromatography","volume":"523","author":"Liu","year":"2004","journal-title":"Anal. Chim. Acta"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1248\/cpb.58.1353","article-title":"Kojic acid derivatives as histamine H3 receptor ligands","volume":"58","author":"Sander","year":"2010","journal-title":"Chem. Pharm. Bull."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Cleal, M., Gibbon, A., Fontana, B.D., and Parker, M.O. (2020). The importance of pH: How aquarium water is affecting behavioural responses to drug exposure in larval zebrafish. Pharmacol. Biochem. Behav., 199.","DOI":"10.1016\/j.pbb.2020.173066"},{"key":"ref_50","first-page":"134","article-title":"Hydroxybenzoic acid derivatives and their biological activity","volume":"23","author":"Safarova","year":"2022","journal-title":"Process. Petrochem. Oil Refin."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Sharma, A., Churungu, D., Giri, A., and Bhardwaj, P. (2024). Cosmeceuticals significance of hydroxybenzoic acids. Specialized Plant Metabolites as Cosmeceuticals, Elsevier.","DOI":"10.1016\/B978-0-443-19148-0.00005-X"},{"key":"ref_52","first-page":"109","article-title":"A comprehensive review on biological activities of p-hydroxy benzoic acid and its derivatives","volume":"22","author":"Manuja","year":"2013","journal-title":"Int. J. Pharm. Sci. Rev. Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1007\/s00403-014-1440-3","article-title":"Effects of resveratrol, oxyresveratrol, and their acetylated derivatives on cellular melanogenesis","volume":"306","author":"Park","year":"2014","journal-title":"Arch. Dermatol. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1007\/s11101-024-09966-4","article-title":"Phytochemical and pharmacological properties of Xanthium species: A review","volume":"24","author":"Wang","year":"2024","journal-title":"Phytochem. Rev."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"115109","DOI":"10.1016\/j.jep.2022.115109","article-title":"TCM herbal prescription recommendation model based on multi-graph convolutional network","volume":"297","author":"Zhao","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1097\/00008390-199412000-00004","article-title":"Melphalan uptake, hyperthermic synergism and drug resistance in a human cell culture model for the isolated limb perfusion of melanoma","volume":"4","author":"Clark","year":"1994","journal-title":"Melanoma Res."},{"key":"ref_57","first-page":"1474","article-title":"Regulation of melanin synthesis of B16 mouse melanoma cells by 1\u03b1, 25-dihydroxyvitamin D3 and retinoic acid","volume":"45","author":"Hosoi","year":"1985","journal-title":"Cancer Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/0006-2952(94)90231-3","article-title":"Reduction of DNA synthesis, pigment synthesis, pigmentation gene mRNA and resistance to UVB in human melanoma cells treated with analogues of a histamine (H2) agonist","volume":"48","author":"Fechner","year":"1994","journal-title":"Biochem. Pharmacol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/18\/3689\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:43:29Z","timestamp":1760035409000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/18\/3689"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,10]]},"references-count":58,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["molecules30183689"],"URL":"https:\/\/doi.org\/10.3390\/molecules30183689","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2025,9,10]]}}}