{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T23:43:02Z","timestamp":1775000582705,"version":"3.50.1"},"reference-count":53,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T00:00:00Z","timestamp":1623110400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/QUI-OUT\/29013\/2017"],"award-info":[{"award-number":["PTDC\/QUI-OUT\/29013\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Tyrosinase is the central enzyme involved in the highly complex process of melanin formation, catalyzing the rate-limiting steps of this biosynthetic pathway. Due to such a preponderant role, it has become a major target in the treatment of undesired skin pigmentation conditions and also in the prevention of enzymatic food browning. Numerous phenolic-based structures from natural sources have been pointed out as potential tyrosinase inhibitors, including anthocyanins. The aim of the present study was to individually assess the tyrosinase inhibitory activity of eight purified compounds with a variable degree of structural complexity: native anthocyanins, deoxyanthocyanins, and pyranoanthocyanins. The latter two, the groups of anthocyanin-related compounds with enhanced stability, were tested for the first time. Compounds 1 to 4 (luteolinidin, deoxymalvidin, cyanidin-, and malvidin-3-O-glucoside) revealed to be the most effective inhibitors, and further kinetic studies suggested their inhibition mechanism to be of a competitive nature. Structure\u2013activity relationships were proposed based on molecular docking studies conducted with mushroom tyrosinase (mTYR) and human tyrosinase-related protein 1 (hTYRP1) crystal structures, providing information about the binding affinity and the different types of interactions established with the enzyme\u2019s active center which corroborated the findings of the inhibition and kinetic studies. Overall, these results support the applicability of these compounds as pigmentation modulators.<\/jats:p>","DOI":"10.3390\/ijms22126192","type":"journal-article","created":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T12:08:55Z","timestamp":1623154135000},"page":"6192","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["The Role of Anthocyanins, Deoxyanthocyanins and Pyranoanthocyanins on the Modulation of Tyrosinase Activity: An In Vitro and In Silico Approach"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2983-4290","authenticated-orcid":false,"given":"Patr\u00edcia","family":"Correia","sequence":"first","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8256-2968","authenticated-orcid":false,"given":"H\u00e9lder","family":"Oliveira","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"given":"Paula","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3130-9807","authenticated-orcid":false,"given":"Nat\u00e9rcia F.","family":"Br\u00e1s","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9779-2678","authenticated-orcid":false,"given":"Ana Rita","family":"Pereira","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"given":"Joana","family":"Moreira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Qu\u00edmica Org\u00e2nica e Farmac\u00eautica, Departamento de Ci\u00eancias Qu\u00edmicas, Faculty of Pharmacy of the University of Porto, Rua Jorge Viterbo Ferreira n\u00ba 228, 4050-313 Porto, Portugal"},{"name":"Centro Interdisciplinar de Investiga\u00e7\u00e3o Marinha e Ambiental (CIIMAR), Universidade do Porto, Edif\u00edcio do Terminal de Cruzeiros do Porto de Leix\u00f5es, Av. General Norton de Matos s\/n, 4050-208 Matosinhos, Portugal"}]},{"given":"Victor","family":"de Freitas","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9318-9732","authenticated-orcid":false,"given":"Nuno","family":"Mateus","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9996-1463","authenticated-orcid":false,"given":"Joana","family":"Oliveira","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2297-0086","authenticated-orcid":false,"given":"Iva","family":"Fernandes","sequence":"additional","affiliation":[{"name":"LAQV-REQUIMTE, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,8]]},"reference":[{"key":"ref_1","first-page":"843","article-title":"Melanocyte biology and skin pigmentation","volume":"445","author":"Lin","year":"2007","journal-title":"Nat. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1600-0749.2005.00288.x","article-title":"Tyrosinase maturation through the mammalian secretory pathway: Bringing color to life","volume":"19","author":"Wang","year":"2006","journal-title":"Pigment. Cell Res."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"D\u2019Mello, S.A.N., Finlay, G.J., Baguley, B.C., and Askarian-Amiri, M.E. (2016). Signaling Pathways in Melanogenesis. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17071144"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1222","DOI":"10.1038\/jid.2011.413","article-title":"Melanosomes Are Transferred from Melanocytes to Keratinocytes through the Processes of Packaging, Release, Uptake, and Dispersion","volume":"132","author":"Ando","year":"2012","journal-title":"J. Investig. Dermatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1111\/ics.12466","article-title":"Intrinsic and extrinsic regulation of human skin melanogenesis and pigmentation","volume":"40","author":"Serre","year":"2018","journal-title":"Int. J. Cosmet. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1002\/pro.2734","article-title":"Structure-function correlations in tyrosinases","volume":"24","author":"Kanteev","year":"2015","journal-title":"Protein Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1111\/j.1755-148X.2009.00636.x","article-title":"New insights into the active site structure and catalytic mechanism of tyrosinase and its related proteins","volume":"22","author":"Olivares","year":"2009","journal-title":"Pigment. Cell Melanoma Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"34993","DOI":"10.1038\/srep34993","article-title":"The unravelling of the complex pattern of tyrosinase inhibition","volume":"6","author":"Deri","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1111\/j.1468-3083.2006.01670.x","article-title":"The safety of hydroquinone","volume":"20","author":"Nordlund","year":"2006","journal-title":"J. Eur. Acad. Dermatol. Venereol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/14756366.2018.1545767","article-title":"A comprehensive review on tyrosinase inhibitors","volume":"34","author":"Zolghadri","year":"2019","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"941","DOI":"10.3109\/13880209.2013.771376","article-title":"Inhibitory effect of liposome-encapsulated anthocyanin on melanogenesis in human melanocytes","volume":"51","author":"Hwang","year":"2013","journal-title":"Pharm. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"127","DOI":"10.3233\/JBR-140075","article-title":"Anti-melanogenic compounds in Rubus croceacanthus","volume":"4","author":"Kubota","year":"2014","journal-title":"J. Berry Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.foodchem.2017.01.041","article-title":"Unraveling the inhibition mechanism of cyanidin-3-sophoroside on polyphenol oxidase and its effect on enzymatic browning of apples","volume":"227","author":"Hemachandran","year":"2017","journal-title":"Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4937","DOI":"10.1021\/acs.chemrev.5b00507","article-title":"Stabilizing and Modulating Color by Copigmentation: Insights from Theory and Experiment","volume":"116","author":"Trouillas","year":"2016","journal-title":"Chem. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1016\/j.foodres.2010.01.022","article-title":"The properties and stability of anthocyanins in mulberry fruits","volume":"43","author":"Aramwit","year":"2010","journal-title":"Food Res. Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4187","DOI":"10.1039\/C7FO01205E","article-title":"Anthocyanin profile, antioxidant activity and enzyme inhibiting properties of blueberry and cranberry juices: A comparative study","volume":"8","author":"Les","year":"2017","journal-title":"Food Funct."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1016\/j.dyepig.2013.09.009","article-title":"Previous and recent advances in pyranoanthocyanins equilibria in aqueous solution","volume":"100","author":"Oliveira","year":"2014","journal-title":"Dyes Pigment."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1111\/1541-4337.12476","article-title":"3-Deoxyanthocyanidin Colorant: Nature, Health, Synthesis, and Food Applications","volume":"18","author":"Xiong","year":"2019","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"12058","DOI":"10.1021\/acsomega.9b01066","article-title":"New Procedure To Calculate All Equilibrium Constants in Flavylium Compounds: Application to the Copigmentation of Anthocyanins","volume":"4","author":"Mendoza","year":"2019","journal-title":"ACS Omega"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.solener.2019.08.050","article-title":"Study of the multi-equilibria of red wine colorants pyranoanthocyanins and evaluation of their potential in dye-sensitized solar cells","volume":"191","author":"Pinto","year":"2019","journal-title":"Sol. Energy"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5106","DOI":"10.1016\/j.tetlet.2013.07.046","article-title":"Network of carboxypyranomalvidin-3-O-glucoside (vitisin A) equilibrium forms in aqueous solution","volume":"54","author":"Oliveira","year":"2013","journal-title":"Tetrahedron Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1538","DOI":"10.1021\/jp110593c","article-title":"Chemical Behavior of Methylpyranomalvidin-3-O-glucoside in Aqueous Solution Studied by NMR and UV\u2212Visible Spectroscopy","volume":"115","author":"Oliveira","year":"2011","journal-title":"J. Phys. Chem. B"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1039\/C1CS15126F","article-title":"Chemistry and applications of flavylium compounds: A handful of colours","volume":"41","author":"Pina","year":"2012","journal-title":"Chem. Soc. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1021\/jf00075a034","article-title":"Color stability and structural transformations of cyanidin 3,5-diglucoside and four 3-deoxyanthocyanins in aqueous solutions","volume":"35","author":"Mazza","year":"1987","journal-title":"J. Agric. Food Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1080\/14756366.2020.1740691","article-title":"Histidine residues at the copper-binding site in human tyrosinase are essential for its catalytic activities","volume":"35","author":"Noh","year":"2020","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.hermed.2018.09.002","article-title":"Validation of medicinal herbs for anti-tyrosinase potential","volume":"14","author":"Mukherjee","year":"2018","journal-title":"J. Herb. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"82","DOI":"10.3136\/fsti9596t9798.3.82","article-title":"Inhibition of Tyrosinase Activity by the Anthocyanin Pigments Isolated from Phaseolus vulgaris L.","volume":"3","author":"Tsuda","year":"1997","journal-title":"Food Sci. Technol. Int. Tokyo"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6011","DOI":"10.1021\/jf200940h","article-title":"Isolation and Tyrosinase Inhibitory Effects of Polyphenols from the Leaves of Persimmon, Diospyros kaki","volume":"59","author":"Xue","year":"2011","journal-title":"J. Agric. Food Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1111\/j.1365-2621.1964.tb00426.x","article-title":"Anthocyanins. VI. Chelation Studies on Anthocyanins and Other Related Compounds","volume":"29","author":"Somaatmadja","year":"1964","journal-title":"J. Food Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4108","DOI":"10.1021\/jf011378z","article-title":"Kinetics of Mushroom Tyrosinase Inhibition by Quercetin","volume":"50","author":"Chen","year":"2002","journal-title":"J. Agric. Food Chem."},{"key":"ref_31","unstructured":"Copeland, R.A. Reversible Modes of Inhibitor Interactions with Enzymes. Evaluation of Enzyme Inhibitors in Drug Discovery 2013, John Wiley & Sons, Inc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"92","DOI":"10.3109\/14756366.2012.755621","article-title":"On the interpretation of tyrosinase inhibition kinetics","volume":"29","author":"Sun","year":"2013","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5232680","DOI":"10.1155\/2017\/5232680","article-title":"Kinetics of Tyrosinase Inhibitory Activity Using Vitis vinifera Leaf Extracts","volume":"2017","author":"Lin","year":"2017","journal-title":"BioMed Res. Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.foodres.2017.07.010","article-title":"Quercetin as a tyrosinase inhibitor: Inhibitory activity, conformational change and mechanism","volume":"100","author":"Fan","year":"2017","journal-title":"Food Res. Int."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3107","DOI":"10.1016\/j.tet.2014.09.051","article-title":"Anthocyanins and derivatives are more than flavylium cations","volume":"71","author":"Pina","year":"2015","journal-title":"Tetrahedron"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4760","DOI":"10.1021\/jf800109a","article-title":"Enzyme, Protein, Carbohydrate, and Phenolic Contaminants in Commercial Tyrosinase Preparations: Potential Problems Affecting Tyrosinase Activity and Inhibition Studies","volume":"56","author":"Flurkey","year":"2008","journal-title":"J. Agric. Food Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.sajb.2018.07.001","article-title":"Cyanidin-3-O-glucoside inhibits different enzymes involved in central nervous system pathologies and type-2 diabetes","volume":"120","author":"Les","year":"2019","journal-title":"S. Afr. J. Bot."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"W665","DOI":"10.1093\/nar\/gkh381","article-title":"PDB2PQR: An automated pipeline for the setup of Poisson-Boltzmann electrostatics calculations","volume":"32","author":"Dolinsky","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"102950","DOI":"10.1016\/j.bioorg.2019.102950","article-title":"Synthesis, computational molecular docking analysis and effectiveness on tyrosinase inhibition of kojic acid derivatives","volume":"88","author":"Karakaya","year":"2019","journal-title":"Bioorg. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.ejmech.2019.06.019","article-title":"Exploiting the 1-(4-fluorobenzyl)piperazine fragment for the development of novel tyrosinase inhibitors as anti-melanogenic agents: Design, synthesis, structural insights and biological profile","volume":"178","author":"Ielo","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1562","DOI":"10.1080\/14756366.2019.1654468","article-title":"Synthesis, computational studies, tyrosinase inhibitory kinetics and antimelanogenic activity of hydroxy substituted 2-[(4-acetylphenyl)amino]-2-oxoethyl derivatives","volume":"34","author":"Rafiq","year":"2019","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1111\/cbdd.13001","article-title":"Inhibition of tyrosinase by 4H -chromene analogs: Synthesis, kinetic studies, and computational analysis","volume":"90","author":"Brasil","year":"2017","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1111\/j.2042-7158.1994.tb03253.x","article-title":"Kojic Acid, a Cosmetic Skin Whitening Agent, is a Slow-binding Inhibitor of Catecholase Activity of Tyrosinase","volume":"46","author":"Cabanes","year":"2011","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"9812","DOI":"10.1002\/anie.201704616","article-title":"Structure of Human Tyrosinase Related Protein 1 Reveals a Binuclear Zinc Active Site Important for Melanogenesis","volume":"56","author":"Lai","year":"2017","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1093\/protein\/8.2.127","article-title":"LIGPLOT: A program to generate schematic diagrams of protein-ligand interactions","volume":"8","author":"Wallace","year":"1995","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_46","first-page":"1270","article-title":"Kinetic characterization of the substrate specificity and mechanism of mushroom tyrosinase","volume":"267","author":"Fenoll","year":"2000","journal-title":"JBIC J. Biol. Inorg. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"4294","DOI":"10.1016\/j.tet.2016.05.076","article-title":"A simple synthesis of 3-deoxyanthocyanidins and their O-glucosides","volume":"72","author":"Mora","year":"2016","journal-title":"Tetrahedron"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3785","DOI":"10.1021\/jf903714z","article-title":"Influence of Anthocyanins, Derivative Pigments and Other Catechol and Pyrogallol-Type Phenolics on Breast Cancer Cell Proliferation","volume":"58","author":"Fernandes","year":"2010","journal-title":"J. Agric. Food Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"6894","DOI":"10.1021\/jf061085b","article-title":"Color Properties of Four Cyanidin\u2212Pyruvic Acid Adducts","volume":"54","author":"Oliveira","year":"2006","journal-title":"J. Agric. Food Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"5477","DOI":"10.1021\/bi200395t","article-title":"Crystal Structure of Agaricus bisporus Mushroom Tyrosinase: Identity of the Tetramer Subunits and Interaction with Tropolone","volume":"50","author":"Ismaya","year":"2011","journal-title":"Biochemistry"},{"key":"ref_51","unstructured":"Dennington, R., Keith, T.A., and Millam, J.M. (2009). GaussView, Version 5, Semichem Inc."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2785","DOI":"10.1002\/jcc.21256","article-title":"AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility","volume":"30","author":"Morris","year":"2009","journal-title":"J. Comput. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0263-7855(96)00018-5","article-title":"VMD: Visual molecular dynamics","volume":"14","author":"Humphrey","year":"1996","journal-title":"J. Mol. Graph."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/12\/6192\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:12:05Z","timestamp":1760163125000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/12\/6192"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,8]]},"references-count":53,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["ijms22126192"],"URL":"https:\/\/doi.org\/10.3390\/ijms22126192","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,8]]}}}