Isolation, bioassay and 3D-QSAR analysis of 8-isopentenyl flavonoids from Epimedium sagittatum Maxim. as PDE5A inhibitors

Isolation, bioassay and 3D-QSAR analysis of 8-isopentenyl flavonoids from Epimedium sagittatum Maxim. as PDE5A inhibitors | 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 Isolation, bioassay and 3D-QSAR analysis of 8-isopentenyl flavonoids from Epimedium sagittatum Maxim. as PDE5A inhibitors Juntao Li, Yue Wu, Xinxin Yu, Xinyu Zheng, Jiechen Xian, Senjie Li, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2300138/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 31 Dec, 2022 Read the published version in Chinese Medicine → Version 1 posted 5 You are reading this latest preprint version Abstract Background: As known, inhibition of phosphodiesterase 5 (PDE5) has the therapeutic effect on male erectile dysfunction (ED), and the processed folium of Epimedium sagittatum Maxim. (PFES) characterized by 8-isopentenyl flavonoids is a famous herb for treating ED. However, the main flavonoids inhibitory activities, structure-activity relationship (SAR) and signaling pathway have been not systematically studied so that its pharmacodynamic mechanism is unclear. Methods: We aimed to initially reveal the PFES efficacy mechanism for treating ED. For the first time, 6 main 8-isopentenyl flavonoids (1-6) from PFES were isolated and identified. Then based on HPLC detection, we proposed a novel method with superior applicability compared to traditional radioisotope assay to screen inhibitors among them. We further established three-dimensional quantitative structure-activity relationship (3D-QSAR) models through CoMFA and CoMSIA to analyze the SAR for those inhibitors. Results: The results were verified by cellular effects of the screened flavonoids. Among 6 compounds, Icariin (1), 2-O"-rhamnosylicaridide II (2) and Baohuoside I (3) were identified with significant activities (IC50 = 8.275, 3.233, 5.473 mM). Then 3D-QSAR studies showed that the replacement of C8 with bulky steric groups as isopentenyl, C3 with positive charge groups and C4' with a hydrogen bond acceptor substituent could increase inhibitory effects. In contrast, the substitution of C7 with bulky steric groups or hydrophilic groups tended to decrease the efficacies. And compounds1, 2, 3 could increase cGMP level and decrease cytoplasmic Ca2+ of rat corpus cavernosum smooth muscle cells (CCSMCs)by activating PKG. Conclusion: 8-isopentenyl flavonoids could be the main pharmacodynamic substances of PFES in the treatment for ED, and some had significant PDE5A1 inhibitory activities so as to activate cGMP/PKG/Ca2+ signaling pathway in CCSMCs, that was related to the substituents at the key sites such as C8, C3, C4' and C7 in the characteristic compounds. 8-isopentenyl flavonoid the processed folium of Epimedium sagittatum Maxim. phosphodiesterase-5 inhibitor 3D-QSAR cGMP-PKG-Ca2+ signaling pathway Full Text Cite Share Download PDF Status: Published Journal Publication published 31 Dec, 2022 Read the published version in Chinese Medicine → Version 1 posted Editorial decision: Major revision 04 Dec, 2022 Reviewers agreed at journal 26 Nov, 2022 Reviewers invited by journal 26 Nov, 2022 Editor assigned by journal 24 Nov, 2022 First submitted to journal 23 Nov, 2022 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. 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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-2300138","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":155201639,"identity":"b4521889-db11-4de0-88e7-90cd989c3be2","order_by":0,"name":"Juntao 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08:18:42","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2300138/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2300138/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s13020-022-00705-5","type":"published","date":"2022-12-31T18:07:38+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":44715224,"identity":"8413a75d-1597-4885-bb8a-14c858fea63b","added_by":"auto","created_at":"2023-10-16 18:13:55","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":902631,"visible":true,"origin":"","legend":"","description":"","filename":"manuscriptsCM.24.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2300138/v1_covered_7d1c3514-109c-4e07-8422-71730b8915d4.pdf"}],"financialInterests":"","formattedTitle":"Isolation, bioassay and 3D-QSAR analysis of 8-isopentenyl flavonoids from Epimedium sagittatum Maxim. as PDE5A inhibitors","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":true,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"chinese-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmed","sideBox":"Learn more about [Chinese Medicine](http://cmjournal.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/cmed/default.aspx","title":"Chinese Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"8-isopentenyl flavonoid, the processed folium of Epimedium sagittatum Maxim., phosphodiesterase-5 inhibitor, 3D-QSAR, cGMP-PKG-Ca2+ signaling pathway","lastPublishedDoi":"10.21203/rs.3.rs-2300138/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2300138/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e\u0026nbsp;As known, inhibition of\u0026nbsp;phosphodiesterase 5\u0026nbsp;(PDE5) has the therapeutic effect on male erectile dysfunction\u0026nbsp;(ED), and the processed folium of Epimedium sagittatum\u0026nbsp;Maxim.\u0026nbsp;(PFES) characterized by 8-isopentenyl flavonoids is\u0026nbsp;a famous\u0026nbsp;herb for treating ED. However, the main flavonoids inhibitory activities, structure-activity relationship\u0026nbsp;(SAR) and signaling pathway\u0026nbsp;have been not systematically studied so that its pharmacodynamic mechanism\u0026nbsp;is unclear.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e We aimed to initially reveal the PFES efficacy mechanism\u0026nbsp;for treating ED. For the first time, 6 main 8-isopentenyl flavonoids (1-6) from PFES were isolated and identified.\u0026nbsp;Then based on HPLC detection, we proposed a novel method with superior applicability compared to traditional radioisotope assay to screen inhibitors among them. We further established three-dimensional quantitative structure-activity relationship\u0026nbsp;(3D-QSAR)\u0026nbsp;models through\u0026nbsp;CoMFA and CoMSIA\u0026nbsp;to analyze the SAR for those\u0026nbsp;inhibitors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u0026nbsp;The\u0026nbsp;results\u0026nbsp;were\u0026nbsp;verified by cellular effects of the screened flavonoids. Among 6 compounds, Icariin (1), 2-O\"-rhamnosylicaridide II (2) and Baohuoside I (3) were identified with significant activities\u0026nbsp;(IC50\u0026nbsp;= 8.275, 3.233, 5.473 mM). Then 3D-QSAR studies showed\u0026nbsp;that\u0026nbsp;the replacement of C8 with bulky steric groups as isopentenyl, C3 with positive charge groups and C4' with a hydrogen bond acceptor substituent could increase inhibitory effects. In contrast, the substitution of C7 with bulky steric groups or hydrophilic groups tended to decrease the efficacies. And compounds1, 2, 3 could\u0026nbsp;increase\u0026nbsp;cGMP level and decrease cytoplasmic Ca2+\u0026nbsp;of rat corpus cavernosum smooth muscle cells (CCSMCs)by activating PKG.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e 8-isopentenyl flavonoids\u0026nbsp;could be the main pharmacodynamic substances of PFES in the treatment for ED, and some had\u0026nbsp;significant PDE5A1 inhibitory activities\u0026nbsp;so as to activate cGMP/PKG/Ca2+ signaling pathway in CCSMCs, that was related to the substituents at the key sites such as C8, C3, C4' and C7 in the characteristic\u0026nbsp;compounds.\u003c/p\u003e","manuscriptTitle":"Isolation, bioassay and 3D-QSAR analysis of 8-isopentenyl flavonoids from Epimedium sagittatum Maxim. as PDE5A inhibitors","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-11-29 04:26:17","doi":"10.21203/rs.3.rs-2300138/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2022-12-04T19:51:34+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2022-11-26T06:33:28+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2022-11-26T06:07:48+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2022-11-24T12:06:23+00:00","index":"","fulltext":""},{"type":"submitted","content":"Chinese Medicine","date":"2022-11-23T19:56:51+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"chinese-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"cmed","sideBox":"Learn more about [Chinese Medicine](http://cmjournal.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/cmed/default.aspx","title":"Chinese Medicine","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2804e370-a859-410e-bb6c-9dc8cc0f61a4","owner":[],"postedDate":"November 29th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-10-16T18:11:52+00:00","versionOfRecord":{"articleIdentity":"rs-2300138","link":"https://doi.org/10.1186/s13020-022-00705-5","journal":{"identity":"chinese-medicine","isVorOnly":false,"title":"Chinese Medicine"},"publishedOn":"2022-12-31 18:07:38","publishedOnDateReadable":"December 31st, 2022"},"versionCreatedAt":"2022-11-29 04:26:17","video":"","vorDoi":"10.1186/s13020-022-00705-5","vorDoiUrl":"https://doi.org/10.1186/s13020-022-00705-5","workflowStages":[]},"version":"v1","identity":"rs-2300138","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2300138","identity":"rs-2300138","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}