{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T14:35:53Z","timestamp":1782484553833,"version":"3.54.5"},"reference-count":30,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2021,11,19]],"date-time":"2021-11-19T00:00:00Z","timestamp":1637280000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81703687"],"award-info":[{"award-number":["81703687"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Science and Technology Research Program of Chongqing Municipal Education Commission","award":["KJQN201801131"],"award-info":[{"award-number":["KJQN201801131"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Tyrosinase is an oxidase that is the rate-limiting enzyme for controlling the production of melanin in the human body. Overproduction of melanin can lead to a variety of skin disorders. Calycosin is an isoflavone from Astragali Radix, which is a traditional Chinese medicine that exhibits several pharmacological activities including skin whitening. In our study, the inhibitory effect of calycosin on melanin production is confirmed in a zebrafish in vivo model by comparing with hydroquinone, kojic acid, and arbutin, known as tyrosinase inhibitors. Moreover, the inhibitory kinetics of calycosin on tyrosinase and their binding mechanisms are determined using molecular docking techniques, molecular dynamic simulations, and free energy analysis. The results indicate that calycosin has an obvious inhibitory effect on zebrafish pigmentation at the concentration of 7.5 \u03bcM, 15 \u03bcM, and 30 \u03bcM. The IC50 of calycosin is 30.35 \u03bcM, which is lower than hydroquinone (37.35 \u03bcM), kojic acid (6.51 \u00d7 103 \u03bcM), and arbutin (3.67 \u00d7 104 \u03bcM). Furthermore, all the results of molecular docking, molecular dynamics simulations, and free energy analysis suggest that calycosin can directly bind to the active site of tyrosinase with very good binding affinity. The study indicates that the combination of computer molecular modeling and zebrafish in vivo assay would be feasible in confirming the result of the in vitro test and illustrating the target-binding information.<\/jats:p>","DOI":"10.3390\/molecules26226998","type":"journal-article","created":{"date-parts":[[2021,11,21]],"date-time":"2021-11-21T21:00:50Z","timestamp":1637528450000},"page":"6998","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Theoretical Exploring of a Molecular Mechanism for Melanin Inhibitory Activity of Calycosin in Zebrafish"],"prefix":"10.3390","volume":"26","author":[{"given":"Nilupaier","family":"Tayier","sequence":"first","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning-Yi","family":"Qin","sequence":"additional","affiliation":[{"name":"Chongqing Pharmaceutical Group Huamosheng Pharmaceutical Science & Technology Co., Ltd., Chongqing 400050, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li-Nan","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Zeng","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3895-1744","authenticated-orcid":false,"given":"Guang","family":"Hu","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"},{"name":"Chongqing Key Laboratory of Medicinal Chemistry & Molecular Pharmacology, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan-Qiang","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, China"},{"name":"Chongqing Key Laboratory of Medicinal Chemistry & Molecular Pharmacology, Chongqing University of Technology, Chongqing 400054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4292","DOI":"10.1158\/0008-5472.CAN-04-4535","article-title":"alpha-Melanocortin and endothelin-1 activate antiapoptotic pathways and reduce DNA damage in human melanocytes","volume":"65","author":"Kadekaro","year":"2005","journal-title":"Cancer Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.fct.2013.12.032","article-title":"Hydroalcoholic extract of Rhodiola rosea L. (Crassulaceae) and its hydrolysate inhibit melanogenesis in B16F0 cells by regulating the CREB\/MITF\/tyrosinase pathway","volume":"65","author":"Chiang","year":"2014","journal-title":"Food Chem. Toxicol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/S0076-6879(87)42024-7","article-title":"Mammalian monophenol monooxygenase (tyrosinase): Purification, properties, and reactions catalyzed","volume":"142","author":"Hearing","year":"1987","journal-title":"Methods Enzymol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1034\/j.1600-0749.2003.00029.x","article-title":"Chemical and instrumental approaches to treat hyperpigmentation","volume":"16","author":"Briganti","year":"2003","journal-title":"Pigment Cell Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5818","DOI":"10.1002\/j.1460-2075.1994.tb06925.x","article-title":"Tyrosinase related protein 1 (TRP1) functions as a DHICA oxidase in melanin biosynthesis","volume":"13","author":"Kobayashi","year":"1994","journal-title":"EMBO J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1007\/s00018-005-5054-y","article-title":"Tyrosinase inhibitors from natural and synthetic sources: Structure, inhibition mechanism and perspective for the future","volume":"62","author":"Kim","year":"2005","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1984","DOI":"10.1271\/bbb.68.1984","article-title":"Tyrosinase inhibitor isolated from the leaves of Zanthoxylum piperitum","volume":"68","author":"Jeong","year":"2004","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1002\/(SICI)1099-1573(199908\/09)13:5<371::AID-PTR453>3.0.CO;2-L","article-title":"Tyrosinase inhibitory p-coumaric acid from ginseng leaves","volume":"13","author":"Lim","year":"1999","journal-title":"Phytother. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1021\/np2007318","article-title":"2-Arylbenzofuran, flavonoid, and tyrosinase inhibitory constituents of Morus yunnanensis","volume":"75","author":"Hu","year":"2012","journal-title":"J. Nat. Prod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1002\/ptr.5344","article-title":"Inhibitory Effects of (2\u2019R)-2\u2019,3\u2019-dihydro-2\u2019-(1-hydroxy-1-methylethyl)-2,6\u2019-bibenzofuran-6,4\u2019-diol on Mushroom Tyrosinase and Melanogenesis in B16-F10 Melanoma Cells","volume":"29","author":"Zhu","year":"2015","journal-title":"Phytother. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1002\/ptr.2435","article-title":"Flavonoids, taxifolin and luteolin attenuate cellular melanogenesis despite increasing tyrosinase protein levels","volume":"22","author":"An","year":"2008","journal-title":"Phytother. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6178","DOI":"10.12659\/MSM.910201","article-title":"Calycosin, a Phytoestrogen Isoflavone, Induces Apoptosis of Estrogen Receptor-Positive MG-63 Osteosarcoma Cells via the Phosphatidylinositol 3-Kinase (PI3K)\/AKT\/Mammalian Target of Rapamycin (mTOR) Pathway","volume":"24","author":"Sun","year":"2018","journal-title":"Med. Sci. Monit."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"744","DOI":"10.3923\/ijp.2019.745.751","article-title":"Calycosin ameliorates inflammatory paw edema in mice via inhibiting NF-kappa B activation","volume":"15","author":"Cui","year":"2019","journal-title":"Int. J. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1002\/cbin.11102","article-title":"Calycosin stimulates the osteogenic differentiation of rat calvarial osteoblasts by activating the IGF1R\/PI3K\/Akt signaling pathway","volume":"43","author":"Fang","year":"2019","journal-title":"Cell Biol. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1002\/ptr.6221","article-title":"Calycosin attenuates MPTP-induced Parkinson\u2019s disease by suppressing the activation of TLR\/NF-\u03baB and MAPK pathways","volume":"33","author":"Yang","year":"2019","journal-title":"Phytother. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1139\/cjpp-2018-0474","article-title":"Inhibition effect of phytoestrogen calycosin on TGF-\u03b21-induced hepatic stellate cell activation, proliferation, and migration via estrogen receptor \u03b2","volume":"96","author":"Deng","year":"2018","journal-title":"Can. J. Physiol. Pharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1248\/bpb.32.264","article-title":"Inhibitory effects of calycosin isolated from the root of Astragalus membranaceus on melanin biosynthesis","volume":"32","author":"Kim","year":"2009","journal-title":"Biol. Pharm. Bull."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1055\/s-2008-1034374","article-title":"Fishing for drugs from nature: Zebrafish as a technology platform for natural product discovery","volume":"74","author":"Crawford","year":"2008","journal-title":"Planta Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1111\/j.1471-4159.2008.05463.x","article-title":"Zebrafish as a new animal model for movement disorders","volume":"106","author":"Flinn","year":"2008","journal-title":"J. Neurochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/2045-824X-4-2","article-title":"Polysaccharides from astragali radix restore chemical-induced blood vessel loss in zebrafish","volume":"4","author":"Hu","year":"2012","journal-title":"Vasc. Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"294","DOI":"10.3109\/00498254.2011.617015","article-title":"Metabolism of calycosin, an isoflavone from Astragali Radix, in zebrafish larvae","volume":"42","author":"Hu","year":"2012","journal-title":"Xenobiotica"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.saa.2016.06.008","article-title":"Inhibitory mechanisms of glabridin on tyrosinase","volume":"168","author":"Chen","year":"2016","journal-title":"Spectrochim. Acta A Mol. Biomol. Spectrosc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.cbi.2015.05.022","article-title":"Modulation effects of sweroside isolated from the Lonicera japonica on melanin synthesis","volume":"238","author":"Jeong","year":"2015","journal-title":"Chem. Biol. Interact."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"105660","DOI":"10.1016\/j.cmpb.2020.105660","article-title":"Molecular dynamics simulation, free energy landscape and binding free energy computations in exploration the anti-invasive activity of amygdalin against metastasis","volume":"195","year":"2020","journal-title":"Comput. Methods Programs Biomed."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/j.foodchem.2018.10.008","article-title":"Identifying an isoflavone from the root of Pueraria lobata as a potent tyrosinase inhibitor","volume":"276","author":"Wagle","year":"2019","journal-title":"Food Chem."},{"key":"ref_26","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_27","doi-asserted-by":"crossref","first-page":"3684","DOI":"10.1063\/1.448118","article-title":"Molecular dynamics with coupling to an external bath","volume":"81","author":"Berendsen","year":"1984","journal-title":"J. Chem. Phys."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1063\/1.445869","article-title":"Comparison of Simple Potential Functions for Simulating Liquid Water","volume":"79","author":"Jorgensen","year":"1983","journal-title":"J. Chem. Phys."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1021\/ar000033j","article-title":"Calculating structures and free energies of complex molecules: Combining molecular mechanics and continuum models","volume":"33","author":"Kollman","year":"2000","journal-title":"Acc. Chem. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1138","DOI":"10.1038\/s41401-019-0217-9","article-title":"Structural insight into the serotonin (5-HT) receptor family by molecular docking, molecular dynamics simulation and systems pharmacology analysis","volume":"40","author":"Wang","year":"2019","journal-title":"Acta Pharmacol. Sin."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/22\/6998\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:33:00Z","timestamp":1760167980000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/26\/22\/6998"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,19]]},"references-count":30,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["molecules26226998"],"URL":"https:\/\/doi.org\/10.3390\/molecules26226998","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,19]]}}}