Structural and mechanistic study of a novel inhibitor of M. tuberculosis cytochrome bc1:aa3

Structural and mechanistic study of a novel inhibitor of M. tuberculosis cytochrome bc1:aa3 | 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 Structural and mechanistic study of a novel inhibitor of M. tuberculosis cytochrome bc1:aa3 Meindert Lamers, Amit Verma, Robbert Kim, Dirk Lamprecht, Clara Aguilar Pérez, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5630670/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Drug-resistant tuberculosis (TB) continues to challenge treatment options, necessitating the exploration of new compounds of novel targets. The mycobacterial respiratory complex cytochrome bc1:aa3 has emerged as a promising target, exemplified by the success of first-in-class inhibitor Q203 in phase 2 clinical trial. However, to fully exploit the potential of this target and to identify the best-in-class inhibitor more compounds need evaluation. Here, we introduce JNJ-2901, a novel inhibitor of the M. tuberculosis cytochrome bc1:aa3, demonstrating activity against multidrug-resistant M. tuberculosis clinical strains at sub-nanomolar concentration and 4-log reduction in bacterial burden in mouse model of TB infection. Inhibitory studies on purified enzyme validate the nanomolar inhibitions observed in mycobacterial cells. Additionally, cryo-EM structure analysis of cytochrome bc1:aa3 bound to JNJ-2901 reveals the binding pocket at the menaquinol oxidation site (Qp), akin to other substate analogue inhibitors like Q203 and TB47. Validation of binding site is further achieved by generating and isolating the JNJ-2901 resistant mutations in M. tuberculosis, followed by purification and resistance analysis of the resistant cytochrome bc1:aa3 complex. Our comprehensive work lay the foundation for further clinical validations of JNJ-2901. Biological sciences/Drug discovery Biological sciences/Drug discovery/Target validation Full Text Additional Declarations Competing interest reported. AKV, RQK, MHL, SW, NV, AB have no competing interests. DL, CAP, JMB, RJC and JW are full-time employees of Janssen, a Johnson & Johnson company, and/or potential stockholders of Johnson & Johnson. JG, CV and DAL have been named inventors in a patent application for JNJ-2901. Supplementary Files Vermacytbcjnj2901SupInf.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 07 Jan, 2025 Reviews received at journal 07 Jan, 2025 Reviewers agreed at journal 05 Jan, 2025 Reviews received at journal 05 Jan, 2025 Reviewers agreed at journal 02 Jan, 2025 Reviewers invited by journal 02 Jan, 2025 Editor assigned by journal 31 Dec, 2024 Submission checks completed at journal 24 Dec, 2024 First submitted to journal 12 Dec, 2024 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. 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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-5630670","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":399164803,"identity":"447d726e-5c3e-447b-ae0e-d8c090f8b1ec","order_by":0,"name":"Meindert 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