Exploring the Therapeutic Potential of Tinospora cordifolia with Computational Insights into EGFR Inhibition

Exploring the Therapeutic Potential of Tinospora cordifolia with Computational Insights into EGFR Inhibition | 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 Research Article Exploring the Therapeutic Potential of Tinospora cordifolia with Computational Insights into EGFR Inhibition Asha G, S Lakshmi, Najmathunnisa N, T Rajva, V. Krishnakumar, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7389657/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 05 Dec, 2025 Read the published version in In Silico Pharmacology → Version 1 posted 13 You are reading this latest preprint version Abstract Tinospora cordifolia ( T. cardifolia ) is a renowned medicinal plant in Ayurveda, known for its immunomodulatory, antioxidant, and anticancer effects. However, integrated studies combining phytochemical, biological, and molecular docking analyses are limited. This study evaluated the antioxidant and cytotoxic activities of T. cordifolia methanolic extract and investigated its major phytoconstituents using density functional theory (DFT) and molecular docking against the EGFR tyrosine kinase domain. The methanolic extract underwent FTIR and UV–Vis spectroscopy to identify functional groups and electronic transitions. Antioxidant activity was tested via DPPH and hydroxyl radical scavenging assays, showing concentration-dependent effects with IC₅₀ values of 235 µg/mL in both assays. Cytotoxicity against EAC and DLA cell lines was assessed by Trypan Blue exclusion and MTT assays. The extract exhibited strong cytotoxicity, particularly on DLA cells (IC₅₀ = 14.3 µg/mL), compared to EAC cells (IC₅₀ = 58.6 µg/mL), with morphological changes indicating apoptosis or necrosis. FTIR revealed N–H, O–H, C–F, and C–Br groups, indicating amines, phenolics, and halogenated compounds. UV–Vis showed a prominent 290 nm absorption related to π→π* transitions of aromatic systems. DFT calculations (B3LYP/6-311 + + G(d,p)) on berberine, palmatine, magnoflorine, and tinocordiside identified Tinocordiside as the most reactive, with the lowest LUMO energy (–1.66 eV), highest electrophilicity index (3.49 eV), and lowest chemical potential (–4.23 eV). Molecular docking against EGFR (PDB ID: 1M17) showed Tinocordiside formed four hydrogen bonds with key residues and had a Glide score of − 6.72 kcal/mol, close to Erlotinib’s − 7.63 kcal/mol. In summary, T. cordifolia methanolic extract exhibits potent antioxidant and anticancer activity, with Tinocordiside as a promising EGFR-targeting lead compound for further therapeutic exploration. Tinospora cordifolia Antioxidant activity Cytotoxicity Density Functional Theory (DFT) Molecular Docking EGFR Inhibition Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 05 Dec, 2025 Read the published version in In Silico Pharmacology → Version 1 posted Editorial decision: Revision requested 07 Sep, 2025 Reviews received at journal 07 Sep, 2025 Reviews received at journal 26 Aug, 2025 Reviews received at journal 26 Aug, 2025 Reviewers agreed at journal 26 Aug, 2025 Reviews received at journal 25 Aug, 2025 Reviewers agreed at journal 25 Aug, 2025 Reviewers agreed at journal 23 Aug, 2025 Reviewers agreed at journal 23 Aug, 2025 Reviewers invited by journal 23 Aug, 2025 Editor assigned by journal 18 Aug, 2025 Submission checks completed at journal 18 Aug, 2025 First submitted to journal 16 Aug, 2025 You are reading this latest preprint version 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-7389657","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":505805891,"identity":"a2da71e4-95e2-4680-85ea-1c445656967e","order_by":0,"name":"Asha G","email":"","orcid":"","institution":"St. Gregorios College","correspondingAuthor":false,"prefix":"","firstName":"Asha","middleName":"","lastName":"G","suffix":""},{"id":505805892,"identity":"03a6df85-1701-4336-9d66-54bfb42ed866","order_by":1,"name":"S Lakshmi","email":"","orcid":"","institution":"Sanatana Dharma College","correspondingAuthor":false,"prefix":"","firstName":"S","middleName":"","lastName":"Lakshmi","suffix":""},{"id":505805894,"identity":"7acf0e4d-4bb7-4999-8f75-8b955313264a","order_by":2,"name":"Najmathunnisa N","email":"","orcid":"","institution":"Farook College","correspondingAuthor":false,"prefix":"","firstName":"Najmathunnisa","middleName":"","lastName":"N","suffix":""},{"id":505805895,"identity":"db78ad39-0204-4561-933a-53bdc306987b","order_by":3,"name":"T Rajva","email":"","orcid":"","institution":"Farook College","correspondingAuthor":false,"prefix":"","firstName":"T","middleName":"","lastName":"Rajva","suffix":""},{"id":505805898,"identity":"f7c886d8-086e-428a-b7ff-dd31d7c055d9","order_by":4,"name":"V. Krishnakumar","email":"","orcid":"","institution":"St. Gregorios College","correspondingAuthor":false,"prefix":"","firstName":"V.","middleName":"","lastName":"Krishnakumar","suffix":""},{"id":505805899,"identity":"581cbe3b-d3c1-41e3-9198-d7d4aea5514f","order_by":5,"name":"K. P. Safna Hussan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5ElEQVRIiWNgGAWjYDAC5gMMBpL/bOQYGNgYGBLAQgcIaGFLYCiwYEsz5iFJy4cKtkOJPSAtRAGDY9yJG27wHEjfz96W+OEBg508A+NZ/NYYHOPdbDhD4k5uD8+xwxIJDMmGDQznEvBrud+7zVjC4Fluj0R6A1ALM1D5GQNCtmz//SfhcDqPRHrzjwSGeqK0bDCQOHA4gUci7RjQlsOEtUiCtEg2pBn2nDmWZpFgcNywjZBf+CBabOTZ29uMb/6oqJbnlyAQYgqo0kAnsUmcwauDQb4BQ4i/B7+WUTAKRsEoGHEAAFbKSbE7m2PwAAAAAElFTkSuQmCC","orcid":"","institution":"Ayurgreen Scientifica Research Institute","correspondingAuthor":true,"prefix":"","firstName":"K.","middleName":"P. Safna","lastName":"Hussan","suffix":""}],"badges":[],"createdAt":"2025-08-16 22:53:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7389657/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7389657/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s40203-025-00468-5","type":"published","date":"2025-12-05T15:58:27+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":97723992,"identity":"fac8dfbf-240e-4088-9fcf-2c0e5761b8c8","added_by":"auto","created_at":"2025-12-08 16:10:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1953974,"visible":true,"origin":"","legend":"","description":"","filename":"20250812Tinosporousmanuscriptadded.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7389657/v1_covered_84de1b12-d115-42e0-a753-c0e1e4cbf28d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Exploring the Therapeutic Potential of Tinospora cordifolia with Computational Insights into EGFR Inhibition","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"in-silico-pharmacology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"insp","sideBox":"Learn more about [In Silico Pharmacology](https://link.springer.com/journal/40203)","snPcode":"40203","submissionUrl":"https://submission.nature.com/new-submission/40203/3","title":"In Silico Pharmacology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Tinospora cordifolia, Antioxidant activity, Cytotoxicity, Density Functional Theory (DFT), Molecular Docking, EGFR Inhibition","lastPublishedDoi":"10.21203/rs.3.rs-7389657/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7389657/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eTinospora cordifolia\u003c/em\u003e (\u003cem\u003eT. cardifolia\u003c/em\u003e) is a renowned medicinal plant in Ayurveda, known for its immunomodulatory, antioxidant, and anticancer effects. However, integrated studies combining phytochemical, biological, and molecular docking analyses are limited. This study evaluated the antioxidant and cytotoxic activities of T. cordifolia methanolic extract and investigated its major phytoconstituents using density functional theory (DFT) and molecular docking against the EGFR tyrosine kinase domain.\u003c/p\u003e\u003cp\u003eThe methanolic extract underwent FTIR and UV\u0026ndash;Vis spectroscopy to identify functional groups and electronic transitions. Antioxidant activity was tested via DPPH and hydroxyl radical scavenging assays, showing concentration-dependent effects with IC₅₀ values of 235 \u0026micro;g/mL in both assays. Cytotoxicity against EAC and DLA cell lines was assessed by Trypan Blue exclusion and MTT assays. The extract exhibited strong cytotoxicity, particularly on DLA cells (IC₅₀ = 14.3 \u0026micro;g/mL), compared to EAC cells (IC₅₀ = 58.6 \u0026micro;g/mL), with morphological changes indicating apoptosis or necrosis. FTIR revealed N\u0026ndash;H, O\u0026ndash;H, C\u0026ndash;F, and C\u0026ndash;Br groups, indicating amines, phenolics, and halogenated compounds. UV\u0026ndash;Vis showed a prominent 290 nm absorption related to π\u0026rarr;π* transitions of aromatic systems. DFT calculations (B3LYP/6-311\u0026thinsp;+\u0026thinsp;+\u0026thinsp;G(d,p)) on berberine, palmatine, magnoflorine, and tinocordiside identified Tinocordiside as the most reactive, with the lowest LUMO energy (\u0026ndash;1.66 eV), highest electrophilicity index (3.49 eV), and lowest chemical potential (\u0026ndash;4.23 eV). Molecular docking against EGFR (PDB ID: 1M17) showed Tinocordiside formed four hydrogen bonds with key residues and had a Glide score of \u0026minus;\u0026thinsp;6.72 kcal/mol, close to Erlotinib\u0026rsquo;s \u0026minus;\u0026thinsp;7.63 kcal/mol.\u003c/p\u003e\u003cp\u003eIn summary, T. cordifolia methanolic extract exhibits potent antioxidant and anticancer activity, with Tinocordiside as a promising EGFR-targeting lead compound for further therapeutic exploration.\u003c/p\u003e","manuscriptTitle":"Exploring the Therapeutic Potential of Tinospora cordifolia with Computational Insights into EGFR Inhibition","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-01 03:39:26","doi":"10.21203/rs.3.rs-7389657/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-07T23:30:56+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-07T16:26:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-26T11:39:04+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-26T07:32:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"74392508706321011829892566732380442148","date":"2025-08-26T07:25:52+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-25T18:33:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"112338043428346544513311132215706373148","date":"2025-08-25T15:56:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"233093217685569668009954862283605705062","date":"2025-08-23T15:14:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"210284237240852590900263582057870811047","date":"2025-08-23T15:13:35+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-08-23T15:06:48+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-08-18T09:32:11+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-18T09:30:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"In Silico Pharmacology","date":"2025-08-16T22:47:43+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"in-silico-pharmacology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"insp","sideBox":"Learn more about [In Silico Pharmacology](https://link.springer.com/journal/40203)","snPcode":"40203","submissionUrl":"https://submission.nature.com/new-submission/40203/3","title":"In Silico Pharmacology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"45d0d725-4304-4617-9049-47066fb1a403","owner":[],"postedDate":"September 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-12-08T16:04:48+00:00","versionOfRecord":{"articleIdentity":"rs-7389657","link":"https://doi.org/10.1007/s40203-025-00468-5","journal":{"identity":"in-silico-pharmacology","isVorOnly":false,"title":"In Silico Pharmacology"},"publishedOn":"2025-12-05 15:58:27","publishedOnDateReadable":"December 5th, 2025"},"versionCreatedAt":"2025-09-01 03:39:26","video":"","vorDoi":"10.1007/s40203-025-00468-5","vorDoiUrl":"https://doi.org/10.1007/s40203-025-00468-5","workflowStages":[]},"version":"v1","identity":"rs-7389657","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7389657","identity":"rs-7389657","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}