Angelic Acid Prevents RANKL-Induced Osteoclastogenesis Through Pathway-Biased Inhibition of MAPK–NFATc1 Signaling

Angelic Acid Prevents RANKL-Induced Osteoclastogenesis Through Pathway-Biased Inhibition of MAPK–NFATc1 Signaling | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Angelic Acid Prevents RANKL-Induced Osteoclastogenesis Through Pathway-Biased Inhibition of MAPK–NFATc1 Signaling Lifang Zhang, Mojtaba Tabandeh, Vishwa Deepak This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8630319/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Excessive osteoclast activity drives inflammatory bone loss in osteoporosis, rheumatoid arthritis, and periodontitis. Natural compounds represent promising therapeutic candidates with favorable safety profiles; however, few exhibit pathway-biased mechanisms of action. Here, we report that angelic acid, a naturally occurring unsaturated monocarboxylic acid, potently inhibits RANKL-induced osteoclastogenesis. This effect occurs with an IC₅₀ of 1.9 µM without cytotoxicity. Mechanistically, angelic acid preferentially suppressed MAPK signaling, including ERK, p38, and JNK phosphorylation. In contrast, NF-κB transcriptional activity remained unaffected, indicating pathway-biased inhibition. Consistent with impaired MAPK activation, angelic acid blocked NFATc1 nuclear translocation and significantly reduced expression of osteoclast-specific genes, including TRAP , cathepsin K , and Atp6v0d2 . These findings identify angelic acid as a novel inhibitor of osteoclast differentiation acting through MAPK-dependent regulation of NFATc1, providing a mechanistic foundation for therapeutic development in bone-resorptive diseases. Drug Discovery, Design, & Development General Cell Biology & Physiology Immunology Angelic acid Osteoclast RANKL MAPK Natural product Bone resorption Full Text Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8630319","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":577304215,"identity":"b46a7454-3b5f-4137-8d4c-25926999f888","order_by":0,"name":"Lifang Zhang","email":"","orcid":"","institution":"Wenzhou-Kean University","correspondingAuthor":false,"prefix":"","firstName":"Lifang","middleName":"","lastName":"Zhang","suffix":""},{"id":577304216,"identity":"23edfc7f-c5ac-416c-b8f3-f2dc1e2b7a3f","order_by":1,"name":"Mojtaba Tabandeh","email":"","orcid":"","institution":"Wenzhou-Kean University","correspondingAuthor":false,"prefix":"","firstName":"Mojtaba","middleName":"","lastName":"Tabandeh","suffix":""},{"id":577304218,"identity":"5f582451-28d4-4da4-989e-469fdc9f342c","order_by":2,"name":"Vishwa Deepak","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAvklEQVRIiWNgGAWjYDCCA4wNzEBKTgLCZSZeizEpWiDKEmcQrYXvRnLz58I2u/SZ7c3PHjBUWCc2ENIieSOxTXpmW3LubJ5j5gYMZ9IJazE4c7CNmXcbc+48iQQzCca2w0Rpaf7Mu60+XU7++TcJxn/EaDne2CDNu+1wgrQED9CWBiK0SB5vbJPm/XfccGZPTplEwrF0Y4Ja+A6zP/7Mc6ZaXuL48W0SH2qsZQlqQQUJpCkfBaNgFIyCUYALAADOHT7xYZ29FgAAAABJRU5ErkJggg==","orcid":"","institution":"Wenzhou-Kean University","correspondingAuthor":true,"prefix":"","firstName":"Vishwa","middleName":"","lastName":"Deepak","suffix":""}],"badges":[],"createdAt":"2026-01-18 09:25:47","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-8630319/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-8630319/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101752531,"identity":"a3c8604a-810c-4006-b9ec-5a0cc6c8e6fa","added_by":"auto","created_at":"2026-02-03 10:28:00","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":476441,"visible":true,"origin":"","legend":"","description":"","filename":"AngelicAcidManuscriptv2.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8630319/v2_covered_4d042434-9f4b-428b-bc9c-13f975088fa3.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"Angelic Acid Prevents RANKL-Induced Osteoclastogenesis Through Pathway-Biased Inhibition of MAPK–NFATc1 Signaling","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Wenzhou-Kean University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Angelic acid, Osteoclast, RANKL, MAPK, Natural product, Bone resorption","lastPublishedDoi":"10.21203/rs.3.rs-8630319/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8630319/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eExcessive osteoclast activity drives inflammatory bone loss in osteoporosis, rheumatoid arthritis, and periodontitis. Natural compounds represent promising therapeutic candidates with favorable safety profiles; however, few exhibit pathway-biased mechanisms of action. Here, we report that angelic acid, a naturally occurring unsaturated monocarboxylic acid, potently inhibits RANKL-induced osteoclastogenesis. This effect occurs with an IC₅₀ of 1.9 \u0026micro;M without cytotoxicity. Mechanistically, angelic acid preferentially suppressed MAPK signaling, including ERK, p38, and JNK phosphorylation. In contrast, NF-κB transcriptional activity remained unaffected, indicating pathway-biased inhibition. Consistent with impaired MAPK activation, angelic acid blocked NFATc1 nuclear translocation and significantly reduced expression of osteoclast-specific genes, including \u003cem\u003eTRAP\u003c/em\u003e, \u003cem\u003ecathepsin K\u003c/em\u003e, and \u003cem\u003eAtp6v0d2\u003c/em\u003e. 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