Characterization and Anticonvulsant Effects of a Colombian Cannabis sativa Extract: A Preclinical Study in Murine Models of Epilepsy

Characterization and Anticonvulsant Effects of a Colombian Cannabis sativa Extract: A Preclinical Study in Murine Models of Epilepsy | 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 Characterization and Anticonvulsant Effects of a Colombian Cannabis sativa Extract: A Preclinical Study in Murine Models of Epilepsy andres david turizo smith This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8935396/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Epilepsy represents one of the most prevalent chronic neurological disorders worldwide, affecting approximately 50 million people globally. Despite the availability of numerous antiseizure medications (ASMs), approximately 30% of patients develop drug-resistant epilepsy, necessitating the exploration of novel therapeutic alternatives. Cannabis sativa has emerged as a promising source of anticonvulsant compounds, particularly cannabidiol (CBD) and Delta-9-tetrahydrocannabinol (THC), which have demonstrated antiepileptic properties in preclinical models. Methods: This study characterized a chloroform extract of Cannabis sativa cultivated in Colombia and evaluated its anticonvulsant potential in three established murine seizure models: the 6 Hz psychomotor seizure model, the pentylenetetrazol (PTZ)-induced seizure model, and the maximal electroshock seizure (MES) test. Phytochemical characterization was performed using thin-layer chromatography (TLC), colorimetric assays (Duquenois-Levine and Fast Blue B Salt), and gas chromatography coupled with mass spectrometry (GCMS/ EI) and flame ionization detection (GC-FID). Results: Chemical analysis revealed a nearly 1:1 ratio of CBD to THC (51% CBD, 49% THC) in the extract, representing an atypical chemotype. In the 6 Hz model, the extract at 100 mg/kg demonstrated complete protection against seizures, comparable to levetiracetam (40 mg/kg). In the MES test, both 50 and 100 mg/kg doses significantly reduced seizure-related mortality. However, in the PTZ model, the extract showed limited efficacy and appeared to exacerbate seizure severity at higher doses, suggesting potential proconvulsant interactions. Conclusions: This study provides evidence for the anticonvulsant potential of a Colombian Cannabis sativa extract in specific seizure models, particularly those involving generalized tonic-clonic and psychomotor seizures. The unique 1:1 CBD:THC ratio suggests possible synergistic interactions between these major cannabinoids. These findings support further investigation of cannabis-based therapies for drug-resistant epilepsy, while highlighting the importance of careful dose optimization and model-specific efficacy considerations. Cellular & Molecular Neuroscience Cannabis sativa cannabidiol delta-9-tetrahydrocannabinol epilepsy anticonvulsant drugresistant epilepsy maximal electroshock pentylenetetrazol 6 Hz seizure model Full Text Additional Declarations The authors declare no competing interests. All animal experimental procedures were reviewed and approved by the Institutional Ethics Committee for Animal Experimentation at the Universidad Nacional de Colombia, in accordance with the guidelines established by the Colombian Ministry of Health (Resolution 008430/1993). The study followed the principles of the 3Rs (Replacement, Reduction, Refinement) for the ethical use of animals in research. Cite Share Download PDF Status: Posted Version 1 posted 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-8935396","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":594967950,"identity":"364b81a7-d636-462b-91e0-baa0756c66b0","order_by":0,"name":"andres david turizo smith","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA70lEQVRIiWNgGAWjYHACAyCWADEYHwAJGagIXmDYANXCDFLKQ6wWMGCTIEoL/+zm7Y9591jY8/OfTqvm3WHHw8DevE2CocYGpxaJO8cKm3meSTBLzsjddpv3TDIPA8+xMgmGY2m4rbmRY9jMc0CCzeAGL1BLGzMPg0SOmQRjw2GcOuShWngMzp/dVszbVs/DIP8GvxYDqBYJgwO525h52w4DbeHBr8XwRlrhzDkHJAyAftksObftOA8bT1qxRQIev8jdSN7w4c2BOmCInd344W1btRw/++GNNz7gCTFMwAYiEkjQMApGwSgYBaMAEwAAEkJKRsBaKuEAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0003-0358-4008","institution":"Universidad Nacional de Colombia","correspondingAuthor":true,"prefix":"","firstName":"andres","middleName":"david turizo","lastName":"smith","suffix":""}],"badges":[],"createdAt":"2026-02-21 19:35:23","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-8935396/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8935396/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103507058,"identity":"7a16f3c6-3a1f-40fb-b839-196b57c20fba","added_by":"auto","created_at":"2026-02-26 13:40:19","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1019737,"visible":true,"origin":"","legend":"","description":"","filename":"preprintCharacterizationandAnticonvulsantEffectsofaColombianCannabissativaExtractAPreclinicalStudyinMurineModelsofEpilepsy.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8935396/v1_covered_a111a6b5-351a-484c-9c74-365ddbdaf136.pdf"}],"financialInterests":"\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003eAll animal experimental procedures were reviewed and approved by the Institutional Ethics Committee for Animal Experimentation at the Universidad Nacional de Colombia, in accordance with the guidelines established by the Colombian Ministry of Health (Resolution 008430/1993). 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Despite the availability of numerous antiseizure medications (ASMs), approximately 30% of patients develop drug-resistant epilepsy, necessitating the exploration of novel therapeutic alternatives. Cannabis sativa has emerged as a promising source of anticonvulsant compounds, particularly cannabidiol (CBD) and Delta-9-tetrahydrocannabinol (THC), which have demonstrated antiepileptic properties in preclinical models.\u003c/p\u003e\n\u003cp\u003eMethods: This study characterized a chloroform extract of Cannabis sativa cultivated in Colombia and evaluated its anticonvulsant potential in three established murine seizure models: the 6 Hz psychomotor seizure model, the pentylenetetrazol (PTZ)-induced seizure model, and the maximal electroshock seizure (MES) test. Phytochemical characterization was performed using thin-layer chromatography (TLC), colorimetric assays (Duquenois-Levine and Fast Blue B Salt), and gas chromatography coupled with mass spectrometry (GCMS/ EI) and flame ionization detection (GC-FID).\u003c/p\u003e\n\u003cp\u003eResults: Chemical analysis revealed a nearly 1:1 ratio of CBD to THC (51% CBD, 49% THC) in the extract, representing an atypical chemotype. In the 6 Hz model, the extract at 100 mg/kg demonstrated complete protection against seizures, comparable to levetiracetam (40 mg/kg). In the MES test, both 50 and 100 mg/kg doses significantly reduced seizure-related mortality. However, in the PTZ model, the extract showed limited efficacy and appeared to exacerbate seizure severity at higher doses, suggesting potential proconvulsant interactions.\u003c/p\u003e\n\u003cp\u003eConclusions: This study provides evidence for the anticonvulsant potential of a Colombian Cannabis sativa extract in specific seizure models, particularly those involving generalized tonic-clonic and psychomotor seizures. The unique 1:1 CBD:THC ratio suggests possible synergistic interactions between these major cannabinoids. These findings support further investigation of cannabis-based therapies for drug-resistant epilepsy, while highlighting the importance of careful dose optimization and model-specific efficacy considerations.\u003c/p\u003e","manuscriptTitle":"Characterization and Anticonvulsant Effects of a Colombian Cannabis sativa Extract: A Preclinical Study in Murine Models of Epilepsy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-25 05:58:09","doi":"10.21203/rs.3.rs-8935396/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":"0ab306bf-53f6-4598-be9e-d4051f160307","owner":[],"postedDate":"February 25th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":63316397,"name":"Cellular \u0026 Molecular Neuroscience"}],"tags":[],"updatedAt":"2026-02-25T05:58:09+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-25 05:58:09","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8935396","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8935396","identity":"rs-8935396","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}