{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T19:10:53Z","timestamp":1785870653273,"version":"3.56.0"},"reference-count":44,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,2,13]],"date-time":"2025-02-13T00:00:00Z","timestamp":1739404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Education","award":["2023RIS-009"],"award-info":[{"award-number":["2023RIS-009"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The investigation of melanogenesis and tyrosinase inhibitors is essential for developing safe and effective natural compounds to treat pigmentation disorders. This study aimed to evaluate the effects of maculosin, a cyclic dipeptide composed of tyrosine and proline, on melanin production and tyrosinase activity using the B16F10 melanoma cell model, while elucidating its mechanism of action through molecular docking and molecular dynamics (MD) simulations. Experimental results demonstrated that maculosin inhibited intracellular melanin content and tyrosinase activity in a concentration-dependent manner in B16F10 melanoma cells. Molecular docking analyses revealed that maculosin exhibited high binding affinities with mushroom tyrosinase (mTYR), tyrosinase-related protein 1 (TYRP1), and Bacillus megaterium tyrosinase (BmTYR) with binding energies of \u22127.7, \u22126.8, and \u22127.5 kcal\/mol, respectively. Furthermore, MD simulations confirmed the structural stability and dynamic flexibility of maculosin\u2013protein complexes, as indicated by RMSD, RMSF, Rg, SASA, hydrogen bond interactions, PCA, and DCCM analyses. Binding free energy calculations using the MM\/PBSA method showed that maculosin exhibited binding energies of \u221228.76 kcal\/mol with mTYR and \u221222.23 kcal\/mol with TYRP1, outperforming standard co-crystal inhibitors such as tropolone (\u221212.47 kcal\/mol) and kojic acid (\u221212.73 kcal\/mol). Critical residues, including VAL-283 and HIS-263 in mTYR and HIS-381, GLY-389, and THR-391 in TYRP1, were identified as key contributors to maculosin binding, corroborating molecular docking findings and displaying strong correlations in DCCM analyses. Collectively, these results suggest that maculosin is a highly promising candidate for the treatment of pigmentation disorders, offering significant inhibitory effects on melanogenesis and tyrosinase activity.<\/jats:p>","DOI":"10.3390\/molecules30040860","type":"journal-article","created":{"date-parts":[[2025,2,13]],"date-time":"2025-02-13T08:01:19Z","timestamp":1739433679000},"page":"860","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["In Vitro and In Silico Studies of Maculosin as a Melanogenesis and Tyrosinase Inhibitor"],"prefix":"10.3390","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-5218-7662","authenticated-orcid":false,"given":"Yang","family":"Xu","sequence":"first","affiliation":[{"name":"Department of Chemistry and Cosmetics, Jeju Inside Agency and Cosmetic Science Center, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8163-1973","authenticated-orcid":false,"given":"Xuhui","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Cosmetics, Jeju Inside Agency and Cosmetic Science Center, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1743-6000","authenticated-orcid":false,"given":"Hyeon-Mi","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Cosmetics, Jeju Inside Agency and Cosmetic Science Center, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3000-7989","authenticated-orcid":false,"given":"Chang-Gu","family":"Hyun","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Cosmetics, Jeju Inside Agency and Cosmetic Science Center, Jeju National University, Jeju 63243, Republic of Korea"},{"name":"Department of Beauty and Cosmetology, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S27","DOI":"10.1016\/j.jaad.2021.12.024","article-title":"Impact of visible light on skin health: The role of antioxidants and free radical quenchers in skin protection","volume":"86","author":"Lim","year":"2022","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Guo, L., Li, W., and Gu, Z. (2023). Recent advances and progress on melanin: From source to application. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24054360"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/s13530-021-00111-8","article-title":"Interaction of various types of bisphenols with enzymes involved in melanin synthesis","volume":"14","author":"Venkatesan","year":"2021","journal-title":"Toxicol. Environ. Health Sci."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Hushcha, Y., Blo, I., and Oton-Gonzalez, L. (2021). microRNAs in the Regulation of Melanogenesis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22116104"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2757","DOI":"10.1111\/jdv.16404","article-title":"Emerging role of dermal compartment in skin pigmentation: Comprehensive review","volume":"34","author":"Kapoor","year":"2020","journal-title":"J. Eur. Acad. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zaidi, M.R., Fisher, D.E., and Rizos, H. (2020). Biology of melanocytes and primary melanoma. Cutaneous Melanoma, Taylor & Francis.","DOI":"10.1007\/978-3-030-05070-2_42"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lee, A.Y. (2021). Skin pigmentation abnormalities and their possible relationship with skin aging. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073727"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.jdermsci.2017.06.018","article-title":"Precise role of dermal fibroblasts on melanocyte pigmentation","volume":"88","author":"Wang","year":"2017","journal-title":"J. Dermatol. Sci."},{"key":"ref_9","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_10","doi-asserted-by":"crossref","first-page":"173","DOI":"10.3892\/etm.2020.8687","article-title":"Natural skin whitening compounds for the treatment of melanogenesis","volume":"20","author":"Qian","year":"2020","journal-title":"Exp. Ther. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2387","DOI":"10.1021\/acschembio.8b00364","article-title":"NRPS protein MarQ catalyzes flexible adenylation and specific S-methylation","volume":"13","author":"Huang","year":"2018","journal-title":"ACS Chem. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1039\/C5NP00035A","article-title":"Insights into the chemical logic and enzymatic machinery of NRPS assembly lines","volume":"33","author":"Walsh","year":"2016","journal-title":"Nat. Prod. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Gondry, M., Jacques, I.B., and Thai, R. (2018). A comprehensive overview of the cyclodipeptide synthase family enriched with the characterization of 32 new enzymes. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00046"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2305","DOI":"10.1039\/C8OB03063D","article-title":"The expanding spectrum of diketopiperazine natural product biosynthetic pathways containing cyclodipeptide synthases","volume":"17","author":"Borgman","year":"2019","journal-title":"Org. Biomol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1038\/s41589-022-01246-6","article-title":"Genome mining for unknown\u2013unknown natural products","volume":"19","author":"Yee","year":"2023","journal-title":"Nat. Chem. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1039\/c2np20010d","article-title":"The nonribosomal synthesis of diketopiperazines in tRNA-dependent cyclodipeptide synthase pathways","volume":"29","author":"Belin","year":"2012","journal-title":"Nat. Prod. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2277","DOI":"10.1007\/s00253-021-11178-1","article-title":"Modifications of diketopiperazines assembled by cyclodipeptide synthases with cytochrome P450 enzymes","volume":"105","author":"Harken","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3574","DOI":"10.1038\/s41467-024-48030-9","article-title":"Cyclodipeptide oxidase is an enzyme filament","volume":"15","author":"Andreas","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1355","DOI":"10.1016\/S1074-5521(02)00285-5","article-title":"The albonoursin gene cluster of S. noursei: Biosynthesis of diketopiperazine metabolites independent of nonribosomal peptide synthetases","volume":"9","author":"Lautru","year":"2002","journal-title":"Chem. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7426","DOI":"10.1073\/pnas.0812191106","article-title":"Identification and structural basis of the reaction catalyzed by CYP121, an essential cytochrome P450 in Mycobacterium tuberculosis","volume":"106","author":"Belin","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7282","DOI":"10.1021\/bi100910y","article-title":"Structural and biochemical characterization of the cytochrome P450 CypX (CYP134A1) from Bacillus subtilis: A cyclo-L-leucyl-L-leucyl dipeptide oxidase","volume":"49","author":"Cryle","year":"2010","journal-title":"Biochemistry"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e02828-17","DOI":"10.1128\/AEM.02828-17","article-title":"Discovery and biosynthesis of the antibiotic bicyclomycin in distantly related bacterial classes","volume":"84","author":"Vior","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1016\/j.chembiol.2013.04.017","article-title":"Insights into the generation of structural diversity in a tRNA-dependent pathway for highly modified bioactive cyclic dipeptides","volume":"20","author":"Giessen","year":"2013","journal-title":"Chem. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"152487","DOI":"10.1016\/j.scitotenv.2021.152487","article-title":"Towards eco-friendly marine antifouling biocides\u2013Nature inspired tetrasubstituted 2, 5-diketopiperazines","volume":"812","author":"Grant","year":"2022","journal-title":"Sci. Total Environ."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Bojarska, J., Mieczkowski, A., and Ziora, Z.M. (2021). Cyclic dipeptides: The biological and structural landscape with special focus on the anti-cancer proline-based scaffold. Biomolecules, 11.","DOI":"10.3390\/biom11101515"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Winyakul, C., Phutdhawong, W., and Tamdee, P. (2022). 2, 5-diketopiperazine derivatives as potential anti-influenza (H5N2) agents: Synthesis, biological evaluation, and molecular docking study. Molecules, 27.","DOI":"10.3390\/molecules27134200"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.2174\/0929867329666220801143650","article-title":"2, 5-Diketopiperazines: A review of source, synthesis, bioactivity, structure, and MS fragmentation","volume":"30","author":"Jia","year":"2023","journal-title":"Curr. Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"891","DOI":"10.2174\/1570193X20666230512162559","article-title":"Research Progress on Activity and Biosynthesis of Diketopiperazines","volume":"21","author":"Liu","year":"2024","journal-title":"Mini-Rev. Org. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1021\/np50116a007","article-title":"New and known diketopiperazines from the Caribbean sponge, Calyx cf. podatypa","volume":"58","author":"Adamczeski","year":"1995","journal-title":"J. Nat. Prod."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"de Moura, G.G.D., de Barros, A.V., and Machado, F. (2021). Endophytic bacteria from strawberry plants control gray mold in fruits via production of antifungal compounds against Botrytis cinerea L.. Microbiol. Res., 251.","DOI":"10.1016\/j.micres.2021.126793"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Xu, Y., Liang, X., and Hyun, C.G. (2024). Isolation, Characterization, Genome Annotation, and Evaluation of Tyrosinase Inhibitory Activity in Secondary Metabolites of Paenibacillus sp. JNUCC32: A Comprehensive Analysis through Molecular Docking and Molecular Dynamics Simulation. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25042213"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1080\/13880209.2021.1946091","article-title":"Maculosin, a non-toxic antioxidant compound isolated from Streptomyces sp. KTM18","volume":"59","author":"Paudel","year":"2021","journal-title":"Pharm. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1142\/S0192415X16500646","article-title":"Inhibitory effect of three diketopiperazines from marine-derived bacteria on HMGB1-induced septic responses in vitro and in vivo","volume":"44","author":"Lee","year":"2016","journal-title":"Am. J. Chin. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1007\/s10593-014-1475-6","article-title":"Cyclo (L-PRO-L-TYR), The fungicide isolated from Lysobacter capsici AZ78: A structure\u2013activity relationship study","volume":"50","author":"Cimmino","year":"2014","journal-title":"Chem. Heterocycl. Compd."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Sekino, Y., Yamamoto, I., and Watanabe, M. (2024). Cyclo (L-Pro-L-Tyr) isolated from the human skin commensal Corynebacterium tuberculostearicum inhibits tyrosinase. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25137365"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Goenka, S., and Simon, R.S. (2020). Inhibitory Effects of the Bioactive Thermorubin Isolated from the Fungus Thermoactinomyces antibioticus on Melanogenesis. Cosmetics, 7.","DOI":"10.3390\/cosmetics7030061"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1687","DOI":"10.1016\/j.bmc.2017.01.037","article-title":"New insights into highly potent tyrosinase inhibitors based on 3-heteroarylcoumarins: Anti-melanogenesis and antioxidant activities, and computational molecular modeling studies","volume":"25","author":"Pintus","year":"2017","journal-title":"Bioorgan. Med. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Nazir, Y., Rafique, H., and Roshan, S. (2022). Molecular docking, synthesis, and tyrosinase inhibition activity of acetophenone amide: Potential inhibitor of melanogenesis. BioMed Res. Int., 2022.","DOI":"10.1155\/2022\/1040693"},{"key":"ref_39","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_40","doi-asserted-by":"crossref","unstructured":"Irfan, A., Faisal, S., and Ahmad, S. (2023). Structure-based virtual screening of furan-1, 3, 4-oxadiazole tethered N-phenylacetamide derivatives as novel class of hTYR and hTYRP1 inhibitors. Pharmaceuticals, 16.","DOI":"10.3390\/ph16030344"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Xu, Y., Liang, X., and Hyun, C.G. (2024). Discovery of Indole\u2013Thiourea Derivatives as Tyrosinase Inhibitors: Synthesis, Biological Evaluation, Kinetic Studies, and In Silico Analysis. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25179636"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1007\/s00894-012-1563-4","article-title":"Principal component and clustering analysis on molecular dynamics data of the ribosomal L11\u00b7 23S subdomain","volume":"19","author":"Wolf","year":"2013","journal-title":"J. Mol. Model."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"204110","DOI":"10.1063\/1.5089636","article-title":"Principal component analysis of nonequilibrium molecular dynamics simulations","volume":"150","author":"Post","year":"2019","journal-title":"J. Chem. Phys."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Meeuwsen, S.M., Hodac, A.N., and Adams, L.M. (2017). Investigation of intrinsic dynamics of enzymes involved in metabolic pathways using coarse-grained normal mode analysis. Cogent Biol., 3.","DOI":"10.1080\/23312025.2017.1291877"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/4\/860\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:32:41Z","timestamp":1760027561000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/4\/860"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,13]]},"references-count":44,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["molecules30040860"],"URL":"https:\/\/doi.org\/10.3390\/molecules30040860","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,13]]}}}