Integrated Stability and Metabolomic Investigation of New Rifampicin and Isoniazid co-loaded Liposome against Tuberculosis

Integrated Stability and Metabolomic Investigation of New Rifampicin and Isoniazid co-loaded Liposome against Tuberculosis | 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 Integrated Stability and Metabolomic Investigation of New Rifampicin and Isoniazid co-loaded Liposome against Tuberculosis PRAGATI SINHA, Vinod L Gaikwad, Debabrata Mandal, E Bhargav, Rahul Laxman Gajbhiye, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7913588/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 This study aimed to develop and evaluate rifampicin (RIF) and isoniazid (INH) co-loaded liposome for sustained drug delivery to enhance therapeutic efficacy against tuberculosis (TB) and overcome challenges associated with prolonged treatment and drug resistance. A novel biocompatible liposomal system that enables sustained co-delivery of RIF and INH, thereby enhancing therapeutic efficacy and reducing drug resistance in TB treatment. However, the formulation offers a stable and scalable delivery platform with sustained release, improved antimicrobial efficacy, and significant potential for advancing tuberculosis therapy. Liposome were prepared using soybean lecithin and cholesterol (L-CH) via rotary evaporator-assisted thin film hydration, optimized by Box–Behnken design, and characterized for size, PDI, entrapment efficiency, and physicochemical properties (FT-IR, DSC, HR-TEM). In vitro release, accelerated stability, antimicrobial efficacy against M. smegmatis and M. tuberculosis H 37 Rv, and LC-MS/MS-based metabolomic profiling were systematically evaluated. The optimised liposome exhibited a mean size of 129.5 ± 2.20 nm, PDI of 0.369 ± 0.06, and entrapment efficiencies of 63.84 ± 1.62% (RIF) and 56.92 ± 1.69% (INH). Following Higuchi diffusion-controlled kinetics, sustained release was observed, with INH (~ 92%) and RIF (~ 85%) over 45 hours. Accelerated stability studies showed minimal drug degradation. Antimicrobial assays revealed a twofold reduction in minimum inhibitory concentration compared to free drugs. Metabolomic analysis demonstrated modulation of glutathione, citric acid, and tyrosine pathways, suggesting improved redox balance and antimicrobial effect. The co-loaded RIF–INH liposomal system offers a promising, clinically translatable approach for sustained drug release and improved tuberculosis therapy. Tuberculosis ⸱ Rifampicin ⸱ Isoniazid ⸱ Liposome ⸱ Metabolomics ⸱ Stability Full Text Additional Declarations No competing interests reported. Supplementary Files SUPPLEMENTARY.docx 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-7913588","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":538919412,"identity":"f2f94273-f14c-4f7d-a528-8495ab10dc26","order_by":0,"name":"PRAGATI SINHA","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"PRAGATI","middleName":"","lastName":"SINHA","suffix":""},{"id":538919413,"identity":"7f2d6992-0d0b-44ac-87d0-2b1ff4ed28ab","order_by":1,"name":"Vinod L 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Tuberculosis","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":"Tuberculosis ⸱ Rifampicin ⸱ Isoniazid ⸱ Liposome ⸱ Metabolomics ⸱ Stability","lastPublishedDoi":"10.21203/rs.3.rs-7913588/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7913588/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study aimed to develop and evaluate rifampicin (RIF) and isoniazid (INH) co-loaded liposome for sustained drug delivery to enhance therapeutic efficacy against tuberculosis (TB) and overcome challenges associated with prolonged treatment and drug resistance. A novel biocompatible liposomal system that enables sustained co-delivery of RIF and INH, thereby enhancing therapeutic efficacy and reducing drug resistance in TB treatment. However, the formulation offers a stable and scalable delivery platform with sustained release, improved antimicrobial efficacy, and significant potential for advancing tuberculosis therapy. Liposome were prepared using soybean lecithin and cholesterol (L-CH) via rotary evaporator-assisted thin film hydration, optimized by Box\u0026ndash;Behnken design, and characterized for size, PDI, entrapment efficiency, and physicochemical properties (FT-IR, DSC, HR-TEM). \u003cem\u003eIn vitro\u003c/em\u003e release, accelerated stability, antimicrobial efficacy against \u003cem\u003eM. smegmatis\u003c/em\u003e and \u003cem\u003eM. tuberculosis\u003c/em\u003e H\u003csub\u003e37\u003c/sub\u003eRv, and LC-MS/MS-based metabolomic profiling were systematically evaluated. The optimised liposome exhibited a mean size of 129.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.20 nm, PDI of 0.369\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06, and entrapment efficiencies of 63.84\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62% (RIF) and 56.92\u0026thinsp;\u0026plusmn;\u0026thinsp;1.69% (INH). Following Higuchi diffusion-controlled kinetics, sustained release was observed, with INH (~\u0026thinsp;92%) and RIF (~\u0026thinsp;85%) over 45 hours. Accelerated stability studies showed minimal drug degradation. Antimicrobial assays revealed a twofold reduction in minimum inhibitory concentration compared to free drugs. Metabolomic analysis demonstrated modulation of glutathione, citric acid, and tyrosine pathways, suggesting improved redox balance and antimicrobial effect. The co-loaded RIF\u0026ndash;INH liposomal system offers a promising, clinically translatable approach for sustained drug release and improved tuberculosis therapy.\u003c/p\u003e","manuscriptTitle":"Integrated Stability and Metabolomic Investigation of New Rifampicin and Isoniazid co-loaded Liposome against Tuberculosis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-05 19:35:49","doi":"10.21203/rs.3.rs-7913588/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":"e54af5e6-8b89-4a1d-8c8f-6f1075d8108c","owner":[],"postedDate":"November 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-14T10:08:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-05 19:35:49","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7913588","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7913588","identity":"rs-7913588","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}