Chalcone Derivative B8HA as A Novel Histone Deacetylase and Tubulin Dual-targeting Inhibitors, Inhibits Triple-Negative Breast Cancer Cells Proliferation

Chalcone Derivative B8HA as A Novel Histone Deacetylase and Tubulin Dual-targeting Inhibitors, Inhibits Triple-Negative Breast Cancer Cells Proliferation | 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 Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Chalcone Derivative B8HA as A Novel Histone Deacetylase and Tubulin Dual-targeting Inhibitors, Inhibits Triple-Negative Breast Cancer Cells Proliferation Yi Su*, Ling Li*, kangkang li, qiuling zheng, shulei qi, yi xie, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1719917/v3 This work is licensed under a CC BY 4.0 License Status: Posted Version 3 posted You are reading this latest preprint version Show more versions Abstract Purpose Designing and synthesizing dual- and multi-target drugs have raised considerable interests due to their advantages in improving potencies as antitumor agents. In previous studies, our group designed and synthesized a series of novel chalcone based tubulin and histone deacetylase (HDAC) dual-targeting inhibitors. Among them, compound B8HA exhibited promising potency for the treatment of triple-negative breast cancer. In this work, we highlighted its biological evaluations in MDA-MB-231 and 4T1 cells. Methods The antitumor activities (including cell cycle arresting and apoptotic effects) and anti-metastasis activities of B8HA were evaluated in vitro . In addition, the potencies of B8HA as inhibitors of tubulin polymerase were also evaluated. Finally, in vivo antitumor and antiangiogenesis effects were explored in 4T1 mice model. Results Comparing to the classical HDAC inhibitor (HDACi) SAHA, B8HA has higher potency to induce apoptosis and inhibits the migratory and invasive abilities of tumor cells under the same dose in vitro and vivo . B8HA as tubulin inhibition is also able to inhibit the formation of capillary-like structures as well as to disrupt existing tubules. Conclusion These results indicated that compound B8HA is a potent inhibitor of both HDAC and tubulin, leading to excellent in vitro and in vivo antitumor activities. Histone deacetylase Tubulin Dual-target inhibitor Triple-negative breast cancer (TNBC) Antiangiogenesis Chalcone Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations The authors declare no competing interests. Supplementary Files graphicalabstract.jpg Graphical abstract B8HA displayed synergic anti-tumor effects due to anti-HDAC and microtubule-destabilizing activities. Scheme.jpg Scheme The design strategy of novel tubulin and HDAC dual-targeting inhibitors. Compounds A and B are two combretastatin-like chalcones displaying potent microtubule-destabilizing activities via acting on the colchicine binding site. The hydrophobic structures make them possible to act as the “cap” part of HDAC inhibitor. We used hydroxamic acid as ZBG due to its high affinity to Zn 2+ . Cite Share Download PDF Status: Posted Version 3 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. 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 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-1719917","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":672524196,"identity":"64a36b0e-542f-467b-92f2-43070da811b8","order_by":0,"name":"Yi Su*","email":"","orcid":"","institution":"Qingdao University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yi","middleName":"","lastName":"Su*","suffix":""},{"id":672524197,"identity":"62dd9d70-2931-47dd-8196-195502c5e1e9","order_by":1,"name":"Ling Li*","email":"","orcid":"","institution":"Qingdao Central Hospital, University of Health and Rehabilitation Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ling","middleName":"","lastName":"Li*","suffix":""},{"id":115606989,"identity":"3e1d50da-f85f-40ce-b249-285b668ef997","order_by":2,"name":"kangkang li","email":"","orcid":"","institution":"Qingdao University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"kangkang","middleName":"","lastName":"li","suffix":""},{"id":115606990,"identity":"3ce6c41a-a46d-4d5e-85c0-6b0afc6e3bed","order_by":3,"name":"qiuling zheng","email":"","orcid":"","institution":"Qingdao University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"qiuling","middleName":"","lastName":"zheng","suffix":""},{"id":115606991,"identity":"0d3bfbff-f9d9-4da4-b75a-971785ed9c06","order_by":4,"name":"shulei qi","email":"","orcid":"","institution":"Qingdao University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"shulei","middleName":"","lastName":"qi","suffix":""},{"id":115606992,"identity":"db16b157-3b05-4f48-b3e1-9631d526ec03","order_by":5,"name":"yi xie","email":"","orcid":"","institution":"Qingdao University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"yi","middleName":"","lastName":"xie","suffix":""},{"id":115606993,"identity":"18795d8c-4faf-45a2-acd6-b45b0ba32fea","order_by":6,"name":"Xuehong Chen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAz0lEQVRIiWNgGAWjYBAC9gYILcfP3tj48AMxWngOQGhjyZ7DzcYSpGhJ3HAjvU2Ahygt7GePSXzcUZs4c+bDNgYJBjs53QZCWnjy0iRnnjlu3C+d2PaggCHZ2OwAAS32DDlm0rxtx2Rnzk5sN5BgOJC4jZAWHv43YC2MG24ebJPgIUqLBNiWGsUNNxiJ1vLG2HJm2wFgICcCA9mACL/w8OcY3vjYVgeMyuMPH36osJMjqAUIWIAReBjKNiCsHASYgcmkjjilo2AUjIJRMDIBAG4iQ3E+in6rAAAAAElFTkSuQmCC","orcid":"","institution":"Qingdao University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Xuehong","middleName":"","lastName":"Chen","suffix":""},{"id":672524198,"identity":"857bead1-a9ac-431f-97ca-1fc2f3c1fa1d","order_by":7,"name":"Yong Fan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7ElEQVRIiWNgGAWjYDACCQiVAMSMDxjYYGwitTAbkKyFTYIoLfyzm4895m2zyeOXbr9W8absMAM/e44Bw88deCy5cyzdcGZbWrHknDNlN+ecO8wg2fPGgLH3DG4tBhI5ZhIf2w4nbriRk3abt+0wg8GNHANmxjZ8WvK/SSS2/QdrKQZpsSesJYcNaMsBoJb0Y8xgWyQIaJG4kWYmOeNccuLMGTnMknPOpfNInHlWcLAXjxb+GcnPpHnK7BL7JdIffnhTZi3H35688cFPPFqQAI8BAw8DCDEwHCBKAwMD+wOI+lEwCkbBKBgFaAAAaUtRuqQu8I4AAAAASUVORK5CYII=","orcid":"","institution":"Department of Psychiatry, Qingdao Mental Health Center","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Yong","middleName":"","lastName":"Fan","suffix":""}],"badges":[],"createdAt":"2022-06-02 14:59:08","currentVersionCode":3,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false,"coiExplicitlySet":true},"doi":"10.21203/rs.3.rs-1719917/v3","doiUrl":"https://doi.org/10.21203/rs.3.rs-1719917/v3","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":114647705,"identity":"d0eaba3d-4cf7-4e22-b88a-03909dff5edf","added_by":"auto","created_at":"2026-07-14 07:17:14","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":8324768,"visible":true,"origin":"","legend":"\u003cp\u003eB8HA inhibit MDA-MD-231 and 4T1 cells proliferation through inducing apoptosis and cell cycle arrest.\u0026nbsp;(A) The morphology of MDA-MB-231 and 4T1 cells treated with different concentrations of B8HA. Images were captured under a bright field at 20x magnification. (B) The investigation of γH2AX in MDA-MB-231 and 4T1 cells by WB. (C) Flow cytometry analysis of apoptosis of MDA-MB-231 and 4T1 cells after treatment with B8HA, SAHA and CA-4. (D) Detection of PARP and caspase-3 quantities in cells by WB. (E) Cell cycle analysis in 4T1 and MDA-MB-231 cells after incubation with different formulations for 24 hours. (F) Cytotoxicity of B8HA against MDA-MB-231 and 4T1 cells at various concentrations for 48 h determined by MTT assay. *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001 compared with control.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/8f1d269ad78c26c14f1c0783.jpg"},{"id":114402611,"identity":"a2988303-a80e-46c0-9330-7a0bd0369343","added_by":"auto","created_at":"2026-07-10 15:03:57","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":66834949,"visible":true,"origin":"","legend":"\u003cp\u003eThe effect of B8HA on migration and invasion of MDA-MB-231 and 4T1 cells was detected by wound healing assay, transwell assay and western blotting.\u0026nbsp;(A, B) MDA-MB-231 and 4T1 cells were treated with 1 and 2 µM of B8HA and 2µM of SAHA for 24 hours, then the migration ability of the cells was detected by wound healing assay and transwell assay. (C) The effect of B8HA on invasion of MDA-MB-231 and 4T1 cells was detected by transwell assay. (D, E) Migration was quantified by measuring the gap closure and the average number of migrating cells. (F) Invasion was quantified by measuring the gap closure and the average number of migrating cells. (G) Detection of MMP-2 and TIMP-2 quantities in cells by WB. *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001 compared with control.\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/ad125006a5a3ab31b16cfc8c.jpg"},{"id":114634802,"identity":"fe0c4c25-b4dd-43a2-9906-08833a77a548","added_by":"auto","created_at":"2026-07-14 05:04:09","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1503714,"visible":true,"origin":"","legend":"\u003cp\u003eThe antiangiogenic and vascular disrupting effects of B8HA were detected by immunofluorescence, tube formation and disruption assay and western blotting.\u003c/p\u003e\n\u003cp\u003e(A) Immunofluorescence assay to detect the effect of B8HA on HUVEC cytoskeleton. The actin cytoskeleton of HUVEC was stained and analyzed by fluorescence confocal imaging after a 16-hour incubation with B8HA and CA-4. Images were captured at 40x magnification. (Scale bar=20 μm) (B) The effect of B8HA on capillary tube formation was observed after a 6-hour incubation under an inverted light microscope. (Scale bar=200 μm) (D) Inhibition rates were evaluated through calculate relative number of tubes. (C) Detection of VEGF quantities in cells by WB. (E) Cell viability of HUVEC cells after B8HA treatment. (G)The effect of B8HA on the disruption of the capillary network was monitored at 0, 1, 2, 3, 6 and 9 hours. (Scale bar=100 μm) (F) Quantitative analysis of the effects of B8HA on the dimensional and topological parameters of the preformed capillary-like tubule networks after a 9 h treatment. *p \u0026lt; 0.05, **p \u0026lt; 0.01, ***P\u0026lt;0.001 compared with control.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/5135a248e37583d5eee4d8ef.png"},{"id":114634462,"identity":"8738843b-43a5-459a-9531-bc9514dd4d7d","added_by":"auto","created_at":"2026-07-14 04:58:19","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":53675446,"visible":true,"origin":"","legend":"\u003cp\u003eTherapeutic efficacy of B8HA in 4T1 mouse model. (A)Tumor volume and (B) tumor weight of the 4T1 tumor-bearing Balb/c mice (n=5) after systemic treatment by saline, 25mg/kg SAHA, 25mg/kg CA-4 and 5, 10, 25 mg/kg B8HAC. (C) Body weight of tumor-bearing Balb/c mice. (D) HE staining and TUNEL staining of tumor tissue. Images were captured at 20x magnification. (Scale bar=200 μm) *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001.\u003c/p\u003e","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/f3c1e9cb7a2673f2067b9464.jpg"},{"id":114402692,"identity":"56ce6660-ed5a-44e9-8be8-f7a263382362","added_by":"auto","created_at":"2026-07-10 15:04:10","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":5290777,"visible":true,"origin":"","legend":"\u003cp\u003e(A, B) Immunohistochemical staining of tumor tissues after treatment with B8HA. Images were captured under a bright field at 10x magnification. (Scale bar=200 μm) (C) H\u0026amp;E staining of mice organs (heart, liver, spleen, lung and kidney) at the end of experiments. (Scale bar=200 μm) *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001.\u003c/p\u003e","description":"","filename":"figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/5be7f34fbcfe9428ac6eaf2a.jpg"},{"id":114401689,"identity":"1b318834-987b-4ee4-add8-417d84049ae0","added_by":"auto","created_at":"2026-07-10 15:02:11","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":309690,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3_covered_e4f2e2f1-d7eb-4a1e-877d-17ea6a3b19c7.pdf"},{"id":114402629,"identity":"68645238-919e-4f0d-950c-63d076863c44","added_by":"auto","created_at":"2026-07-10 15:03:59","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1399669,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical abstract \u003c/strong\u003eB8HA displayed synergic anti-tumor effects due to anti-HDAC and microtubule-destabilizing activities.\u003c/p\u003e","description":"","filename":"graphicalabstract.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/8b80bc83e8886d30bd34651c.jpg"},{"id":114402623,"identity":"c351f5f1-8032-40ab-879c-638b7e7049b5","added_by":"auto","created_at":"2026-07-10 15:03:59","extension":"jpg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1318751,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme \u003c/strong\u003eThe design strategy of novel tubulin and HDAC dual-targeting inhibitors. Compounds A and B are two combretastatin-like chalcones displaying potent microtubule-destabilizing activities via acting on the colchicine binding site. The hydrophobic structures make them possible to act as the “cap” part of HDAC inhibitor. We used hydroxamic acid as ZBG due to its high affinity to Zn\u003csup\u003e2+\u003c/sup\u003e.\u003c/p\u003e","description":"","filename":"Scheme.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1719917/v3/14743d5ee811556a8338ad83.jpg"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"Chalcone Derivative B8HA as A Novel Histone Deacetylase and Tubulin Dual-targeting Inhibitors, Inhibits Triple-Negative Breast Cancer Cells Proliferation","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","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":"Histone deacetylase, Tubulin, Dual-target inhibitor, Triple-negative breast cancer (TNBC), Antiangiogenesis, Chalcone","lastPublishedDoi":"10.21203/rs.3.rs-1719917/v3","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1719917/v3","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose \u003c/strong\u003eDesigning and synthesizing dual- and multi-target drugs have raised considerable interests due to their advantages in improving potencies as antitumor agents. In previous studies, our group designed and synthesized a series of novel chalcone based tubulin and histone deacetylase (HDAC) dual-targeting inhibitors. Among them, compound B8HA exhibited promising potency for the treatment of triple-negative breast cancer. In this work, we highlighted its biological evaluations in MDA-MB-231 and 4T1 cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods \u003c/strong\u003eThe antitumor activities (including cell cycle arresting and apoptotic effects) and anti-metastasis activities of B8HA were evaluated \u003cem\u003ein vitro\u003c/em\u003e. In addition, the potencies of B8HA as inhibitors of tubulin polymerase were also evaluated. Finally, \u003cem\u003ein vivo\u003c/em\u003e antitumor and antiangiogenesis effects were explored in 4T1 mice model.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults \u003c/strong\u003eComparing to the classical HDAC inhibitor (HDACi) SAHA, B8HA has higher potency to induce apoptosis and inhibits the migratory and invasive abilities of tumor cells under the same dose\u003cem\u003e in vitro and vivo\u003c/em\u003e. B8HA as tubulin inhibition is also able to inhibit the formation of capillary-like structures as well as to disrupt existing tubules.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion \u003c/strong\u003eThese results indicated that compound B8HA is a potent inhibitor of both HDAC and tubulin, leading to excellent \u003cem\u003ein vitro and in vivo\u003c/em\u003e antitumor activities.\u003c/p\u003e","manuscriptTitle":"Chalcone Derivative B8HA as A Novel Histone Deacetylase and Tubulin Dual-targeting Inhibitors, Inhibits Triple-Negative Breast Cancer Cells Proliferation","msid":"","msnumber":"","nonDraftVersions":[{"code":3,"date":"2026-07-08 21:32:34","doi":"10.21203/rs.3.rs-1719917/v3","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","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}},{"code":2,"date":"2022-06-22 15:07:16","doi":"10.21203/rs.3.rs-1719917/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","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}},{"code":1,"date":"2022-06-07 16:11:21","doi":"10.21203/rs.3.rs-1719917/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","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}}],"origin":"","ownerIdentity":"98120417-a754-41ea-9e9b-e3e0f0f4703d","owner":[],"postedDate":"July 8th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-09-12T11:36:53+00:00","versionOfRecord":[],"versionCreatedAt":"2026-07-08 21:32:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v3","identity":"rs-1719917","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1719917","identity":"rs-1719917","version":["v3"]},"buildId":"PRX_uewTKcrkEM4liVh3c","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}