Rapid identification of EphA2 ligand-binding domain binders through an optimized NMR fragment-screening workflow

Rapid identification of EphA2 ligand-binding domain binders through an optimized NMR fragment-screening workflow | 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 Article Rapid identification of EphA2 ligand-binding domain binders through an optimized NMR fragment-screening workflow Harald Schwalbe, Sattar Moghaddam, Konstantin Mineev, Annika Wenzel, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8901466/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 Eph receptors are involved in the regulation of cell adhesion and migration and are implicated in cancer progression, making them important drug targets. To date, the design of drugs targeting the ligand-binding domain of Eph includes the development of peptide mimetics of the ephrin ligands and the optimization of repurposed drugs. In this work, we report the results of a fragment-based screening (FBS) campaign against the ligand-binding domain of the EphA2 receptor. We introduce the workflow for the selection, filtering and follow-up studies of FBS hits, including an NMR/X-ray hybrid approach for the structure determination of protein/ligand complexes. The proposed workflow allowed us to identify several compounds with the receptor binding affinities of 50-100 µM and IC 50 of 1 µM , better than the activities of the known repurposed drugs. Due to the low molecular weight, the newly developed hits that exhibit an initial biological effect have a high potential for further optimization. Biological sciences/Structural biology/NMR spectroscopy/Solution-state NMR Biological sciences/Chemical biology/Screening Full Text Additional Declarations There is NO Competing Interest. Supplementary Files EphA2LBDpaperfinalsupplementary.pdf Supplmentary data EphA2LBDpaperfinalsupplementarynewtitle.pdf Supplementary Info File #1 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-8901466","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":593342336,"identity":"73c27e61-9241-4a8f-9a3b-b32bfa379839","order_by":0,"name":"Harald 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