{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T09:09:35Z","timestamp":1764061775588,"version":"3.45.0"},"reference-count":48,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,11,23]],"date-time":"2025-11-23T00:00:00Z","timestamp":1763856000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Regional Innovation System & Education"},{"name":"Ministry of Education (MOE) and the Jeju Special Self-Governing Province","award":["2025-RISE-17-001"],"award-info":[{"award-number":["2025-RISE-17-001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Coumarin-based compounds exhibit diverse pharmacological properties, and 4-methylcoumarin (4MC) has emerged as a promising scaffold for drug development. However, its anti-obesity mechanisms remain insufficiently understood. This study aimed to evaluate the anti-adipogenic potential of 4MC derivatives in 3T3-L1 preadipocytes and to elucidate their underlying molecular mechanisms. Methods: 3T3-L1 preadipocytes were treated with structurally diverse 4MC derivatives. Lipid accumulation was analyzed using Oil Red O staining, cell viability by MTT assay, and the expression of adipogenic proteins by Western blotting. Molecular docking and molecular dynamics simulations were performed to predict the interactions between lead compounds and key adipogenic regulators. Results: Among the tested derivatives, 6,7-dihydroxy-4-methylcoumarin (6,7DH-4MC) markedly inhibited lipid accumulation in a dose-dependent manner without cytotoxicity. It suppressed the expression of major adipogenic transcription factors (PPAR-\u03b3, C\/EBP\u03b1, SREBP-1c) and FABP4. Additionally, 6,7DH-4MC inhibited ERK1\/2 and p38 MAPK phosphorylation while activating AMPK. It also reduced CREB phosphorylation, indicating suppression of early adipogenesis. Computational analyses revealed stable binding of 6,7DH-4MC within the active sites of multiple adipogenic regulators, supporting its pleiotropic mode of action. Conclusions: 6,7DH-4MC exerts potent anti-adipogenic effects by modulating key adipogenic signaling pathways and transcriptional networks. These findings highlight 6,7DH-4MC as a promising lead compound for anti-obesity drug development, warranting further in vivo studies.<\/jats:p>","DOI":"10.3390\/ph18121780","type":"journal-article","created":{"date-parts":[[2025,11,24]],"date-time":"2025-11-24T10:24:17Z","timestamp":1763979857000},"page":"1780","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["6,7-Dihydroxy-4-methylcoumarin Suppresses Adipogenesis via AMPK and MAPK Signaling with In Silico Analysis of Adipogenic Proteins"],"prefix":"10.3390","volume":"18","author":[{"given":"Ye-Jin","family":"Lee","sequence":"first","affiliation":[{"name":"Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Xu","sequence":"additional","affiliation":[{"name":"Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3000-7989","authenticated-orcid":false,"given":"Chang-Gu","family":"Hyun","sequence":"additional","affiliation":[{"name":"Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1002\/oby.23444","article-title":"Metabolic dysfunction and obesity-related cancer: Beyond obesity and metabolic syndrome","volume":"30","author":"Karra","year":"2022","journal-title":"Obesity"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"332","DOI":"10.3389\/fendo.2020.00332","article-title":"Health Consequences of Sarcopenic Obesity: A Narrative Review","volume":"11","author":"Roh","year":"2020","journal-title":"Front. Endocrinol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.ecl.2021.05.013","article-title":"An Update on the Epidemiology of Type 2 Diabetes: A Global Perspective","volume":"50","author":"Tinajero","year":"2021","journal-title":"Endocrinol. Metab. Clin. North. Am."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e13535","DOI":"10.1111\/obr.13535","article-title":"Economic burden of childhood overweight and obesity: A systematic review and meta-analysis","volume":"24","author":"Ling","year":"2023","journal-title":"Obes. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1186\/s12955-022-01974-2","article-title":"Mental health and quality of life in different obesity phenotypes: A systematic review","volume":"20","author":"Abiri","year":"2022","journal-title":"Health Qual. Life Outcomes"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1186\/s12889-021-12449-2","article-title":"Burden of disease study of overweight and obesity; the societal impact in terms of cost-of-illness and health-related quality of life","volume":"22","author":"Hecker","year":"2022","journal-title":"BMC Public Health"},{"key":"ref_7","first-page":"e45518","article-title":"The Effectiveness of Nutritional Strategies in the Treatment and Management of Obesity: A Systematic Review","volume":"15","author":"Adeola","year":"2023","journal-title":"Cureus"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"S288","DOI":"10.37765\/ajmc.2022.89292","article-title":"A review of current guidelines for the treatment of obesity","volume":"28","author":"Cornier","year":"2022","journal-title":"Am. J. Manag. Care"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e34837","DOI":"10.1016\/j.heliyon.2024.e34837","article-title":"Pharmacovigilance analysis of orlistat adverse events based on the FDA adverse event reporting system (FAERS) database","volume":"10","author":"Zhu","year":"2024","journal-title":"Heliyon"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1611","DOI":"10.1007\/s00228-021-03160-7","article-title":"Cardiovascular safety of long-term anti-obesity drugs in subjects with overweight or obesity: A systematic review and meta-analysis","volume":"77","author":"Zhang","year":"2021","journal-title":"Eur. J. Clin. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Gjermeni, E., Kirstein, A.S., Kolbig, F., Kirchhof, M., Bundalian, L., Katzmann, J.L., Laufs, U., Bl\u00fcher, M., Garten, A., and Le Duc, D. (2021). Obesity\u2014An Update on the Basic Pathophysiology and Review of Recent Therapeutic Advances. Biomolecules, 11.","DOI":"10.3390\/biom11101426"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"100024","DOI":"10.1016\/j.obpill.2022.100024","article-title":"Obesity pillars roundtable: Phentermine\u2014Past, present, and future","volume":"3","author":"Bays","year":"2022","journal-title":"Obes. Pillars"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Nevola, R., Epifani, R., Imbriani, S., Tortorella, G., Aprea, C., Galiero, R., Rinaldi, L., Marfella, R., and Sasso, F.C. (2023). GLP-1 Receptor Agonists in Non-Alcoholic Fatty Liver Disease: Current Evidence and Future Perspectives. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021703"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"114405","DOI":"10.1016\/j.ejmech.2022.114405","article-title":"Recent advances in natural anti-obesity compounds and derivatives based on in vivo evidence: A mini-review","volume":"237","author":"Chang","year":"2022","journal-title":"Eur. J. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1080\/13543776.2021.1954161","article-title":"Mechanistically acting anti-obesity compositions\/formulations of natural origin: A patent review (2010\u20132021)","volume":"32","author":"Sengupta","year":"2022","journal-title":"Expert. Opin. Ther. Pat."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1007\/s11033-020-06036-8","article-title":"Molecular mechanism of down-regulating adipogenic transcription factors in 3T3-L1 adipocyte cells by bioactive anti-adipogenic compounds","volume":"48","author":"Guru","year":"2021","journal-title":"Mol. Biol. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Ruiz-Ojeda, F.J., Rup\u00e9rez, A.I., Gomez-Llorente, C., Gil, A., and Aguilera, C.M. (2016). Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17071040"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"120422","DOI":"10.1016\/j.lfs.2022.120422","article-title":"Comparing the effects of tumor necrosis factor alpha, lipopolysaccharide and palmitic acid on lipid metabolism and inflammation in murine 3T3-L1 adipocytes","volume":"297","author":"Jack","year":"2022","journal-title":"Life Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1016\/j.biochi.2010.04.017","article-title":"Antioxidant properties of 4-methylcoumarins in in vitro cell-free systems","volume":"92","author":"Morabito","year":"2010","journal-title":"Biochimie"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.biochi.2017.07.010","article-title":"Protective effects of 4-methylcoumarins and related compounds as radical scavengers and chain-breaking antioxidants","volume":"140","author":"Kancheva","year":"2017","journal-title":"Biochimie"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Raunio, H., Pentik\u00e4inen, O., and Juvonen, R.O. (2020). Coumarin-Based Profluorescent and Fluorescent Substrates for Determining Xenobiotic-Metabolizing Enzyme Activities In Vitro. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21134708"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Lee, Y.J., and Hyun, C.G. (2024). Mechanistic Insights into the Stimulatory Effect of Melanogenesis of 4-Methylcoumarin Derivatives in B16F10 Melanoma Cells. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252212421"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1248\/bpb.b13-00568","article-title":"4-Methylcoumarin derivatives with anti-inflammatory effects in activated microglial cells","volume":"37","author":"Togna","year":"2014","journal-title":"Biol. Pharm. Bull."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2543","DOI":"10.1093\/jac\/dkr355","article-title":"Characterization of 7-amino-4-methylcoumarin as an effective antitubercular agent: Structure-activity relationships","volume":"66","author":"Tandon","year":"2011","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s00253-007-1305-1","article-title":"Antimicrobial activity of an endophytic Xylaria sp. YX-28 and identifi-cation of its antimicrobial compound 7-amino-4-methylcoumarin","volume":"78","author":"Liu","year":"2008","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1007\/s12640-014-9507-7","article-title":"7,8-Dihydroxy-4-methylcoumarin provides neuroprotection by increasing hippocalcin expression","volume":"27","author":"Jin","year":"2015","journal-title":"Neurotox. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2681","DOI":"10.1016\/j.biochi.2012.09.006","article-title":"In vitro platelet antiaggregatory properties of 4-methylcoumarins","volume":"94","author":"Prasad","year":"2012","journal-title":"Biochimie"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"155563","DOI":"10.1016\/j.phymed.2024.155563","article-title":"Epigallocatechin gallate suppresses mitotic clonal expansion and adipogenic differentiation of preadipocytes through impeding JAK2\/STAT3-mediated transcriptional cascades","volume":"12","author":"Peng","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"12272","DOI":"10.1021\/acs.jafc.5c00973","article-title":"Green Tea Epigallocatechin-3-gallate Ameliorates Lipid Accumulation and Obesity-Associated Metabolic Syndrome via Regulating Autophagy and Lipolysis in Preadipocytes and Adipose Tissue","volume":"73","author":"Peng","year":"2025","journal-title":"J. Agric. Food Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"117073","DOI":"10.1016\/j.biopha.2024.117073","article-title":"Novel insights into regulators and functional modulators of adipogenesis","volume":"177","author":"Kim","year":"2024","journal-title":"Biomed. Pharmacother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.7150\/ijbs.27173","article-title":"The PI3K\/AKT pathway in obesity and type 2 diabetes","volume":"14","author":"Huang","year":"2018","journal-title":"Int. J. Biol. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1007\/s13197-022-05436-x","article-title":"Resveratrol butyrate esters inhibit lipid biosynthesis in 3T3-L1 cells by AMP-activated protein kinase phosphorylation","volume":"60","author":"Shih","year":"2023","journal-title":"J. Food Sci. Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.2217\/pgs.11.44","article-title":"Transporter-mediated drug\u2013drug interactions","volume":"12","author":"Fromm","year":"2011","journal-title":"Pharmacogenomics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"279","DOI":"10.2165\/00003088-200544030-00005","article-title":"Mechanism-based inhibition of cytochrome P450 3A4 by therapeutic drugs","volume":"44","author":"Zhou","year":"2005","journal-title":"Clin. Pharmacokinet."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"343","DOI":"10.18388\/pb.2016_35","article-title":"Interactions of CCAAT\/enhancer-binding protein \u03b2 with transcriptional coregulators","volume":"62","author":"Miller","year":"2016","journal-title":"Post\u0119py Biochemii."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1111\/j.1742-4658.2008.06807.x","article-title":"SREBPs: Protein interaction and SREBPs","volume":"276","author":"Sato","year":"2009","journal-title":"FEBS J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1231006","DOI":"10.3389\/fphar.2023.1231006","article-title":"Bioactivity and toxicity of coumarins from African medicinal plants","volume":"14","author":"Anywar","year":"2024","journal-title":"Front. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"El-Sawy, E.R., Abdel-Aziz, M.S., Abdelmegeed, H., and Kirsch, G. (2024). Coumarins: Quorum Sensing and Biofilm Formation Inhibition. Molecules, 29.","DOI":"10.3390\/molecules29194534"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"100202","DOI":"10.1016\/j.crphar.2024.100202","article-title":"Coumarin: A natural solution for alleviating inflammatory disorders","volume":"7","author":"Saadati","year":"2024","journal-title":"Curr. Res. Pharmacol. Drug Discov."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"22791","DOI":"10.1038\/s41598-023-50170-9","article-title":"Synthesis and in-vitro anti-proliferative with antimicrobial activity of new coumarin containing heterocycles hybrids","volume":"13","author":"Abdelaziz","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1362992","DOI":"10.3389\/fchem.2024.1362992","article-title":"Syntheses, reactivity, and biological applications of coumarins","volume":"12","author":"Citarella","year":"2024","journal-title":"Front. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Rohman, N., Ardiansah, B., Wukirsari, T., and Judeh, Z. (2024). Recent Trends in the Synthesis and Bioactivity of Coumarin, Coumarin\u2013Chalcone, and Coumarin\u2013Triazole Molecular Hybrids. Molecules, 29.","DOI":"10.3390\/molecules29051026"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/nm.3159","article-title":"PPAR\u03b3 signaling and metabolism: The good, the bad and the future","volume":"19","author":"Ahmadian","year":"2013","journal-title":"Nat. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1038\/s42003-021-02735-5","article-title":"Activation of the adipocyte CREB\/CRTC pathway in obesity","volume":"4","author":"Yoon","year":"2021","journal-title":"Commun. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Lim, S.H., Lee, H.S., Han, H.K., and Cho, C.I. (2021). Saikosaponin A and D Inhibit Adipogenesis via the AMPK and MAPK Signaling Pathways in 3T3-L1 Adipocytes. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222111409"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"120131","DOI":"10.1016\/j.lfs.2021.120131","article-title":"Quinizarin suppresses the differentiation of adipocytes and lipogenesis in vitro and in vivo via downregulation of C\/EBP-beta\/SREBP pathway","volume":"287","author":"Ree","year":"2021","journal-title":"Life Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"970292","DOI":"10.3389\/fphys.2022.970292","article-title":"The AMPK pathway in fatty liver disease","volume":"13","author":"Fang","year":"2022","journal-title":"Front. Physiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1016\/j.molcel.2017.05.032","article-title":"AMPK: Mechanisms of Cellular Energy Sensing and Restoration of Metabolic Balance","volume":"66","author":"Garcia","year":"2017","journal-title":"Mol. 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