Design of allosteric modulators that change GPCR G protein subtype selectivity

Design of allosteric modulators that change GPCR G protein subtype selectivity | 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 Biological Sciences - Article Design of allosteric modulators that change GPCR G protein subtype selectivity Lauren Slosky, Madelyn Moore, Kelsey Person, Abigail Alwin, Campbell Krusemark, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5538058/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Oct, 2025 Read the published version in Nature → Version 1 posted You are reading this latest preprint version Abstract G protein-coupled receptors (GPCRs), the largest family of drug targets, can signal through 16 subtypes of Gα proteins. Biased compounds that selectively activate therapy-relevant pathways promise to be safer, more effective medications. The determinants of bias are poorly understood, however, and rationally-designed, G protein-subtype-selective compounds are lacking. Here, using the prototypical class A GPCR neurotensin receptor 1 (NTSR1), we find that small molecules binding the intracellular GPCR-transducer interface change G protein coupling by subtype-specific and predictable mechanisms, enabling rational drug design. We demonstrate that the compound SBI-553 switches NTSR1 G protein preference by acting both as a molecular bumper and a molecular glue. Structurally, SBI-553 occludes G protein binding determinants on NTSR1, promoting association with select G protein subtypes for which an alternative, shallow-binding conformation is energetically favorable. Minor modifications to the SBI-553 scaffold produce allosteric modulators with distinct G protein subtype selectivity profiles. Selectivity profiles are probe-independent, conserved across species, and translate to differences in in vivo activity. These studies demonstrate that G protein selectivity can be tailored with small changes to a single chemical scaffold targeting the receptor-transducer interface and, as this pocket is broadly conserved, present a strategy for pathway-selective drug discovery applicable to the diverse GPCR superfamily. Biological sciences/Chemical biology/Pharmacology/Receptor pharmacology Biological sciences/Drug discovery/Pharmacology/Receptor pharmacology G protein-coupled receptor biased signaling β-arrestin G protein selectivity allosteric modulation neurotensin receptor 1 Full Text Additional Declarations Yes there is potential Competing Interest. US Patents 9,868,707 and 20,150,329,497 relating to the chemistry of SBI-553 and its derivatives have been issued to the Sanford Burnham Prebys Medical Research Institute (SBP) and Duke University and US Patent 10,118,902 has been issued to SBP. Patent application 63/689,904 related to the use of SBI-553 and its derivatives has been filed by Duke University. Supplementary Files SupplementaryMaterial1124Final.pdf Supplementary Information Cite Share Download PDF Status: Published Journal Publication published 22 Oct, 2025 Read the published version in Nature → 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. 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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-5538058","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":388539952,"identity":"f5ac5d31-7878-4a8e-963b-943a367c4bdb","order_by":0,"name":"Lauren 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Biased compounds that selectively activate therapy-relevant pathways promise to be safer, more effective medications. The determinants of bias are poorly understood, however, and rationally-designed, G protein-subtype-selective compounds are lacking. Here, using the prototypical class A GPCR neurotensin receptor 1 (NTSR1), we find that small molecules binding the intracellular GPCR-transducer interface change G protein coupling by subtype-specific and predictable mechanisms, enabling rational drug design. We demonstrate that the compound SBI-553 switches NTSR1 G protein preference by acting both as a molecular bumper and a molecular glue. Structurally, SBI-553 occludes G protein binding determinants on NTSR1, promoting association with select G protein subtypes for which an alternative, shallow-binding conformation is energetically favorable. Minor modifications to the SBI-553 scaffold produce allosteric modulators with distinct G protein subtype selectivity profiles. Selectivity profiles are probe-independent, conserved across species, and translate to differences in in vivo activity. These studies demonstrate that G protein selectivity can be tailored with small changes to a single chemical scaffold targeting the receptor-transducer interface and, as this pocket is broadly conserved, present a strategy for pathway-selective drug discovery applicable to the diverse GPCR superfamily.","manuscriptTitle":"Design of allosteric modulators that change GPCR G protein subtype selectivity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-11 06:59:30","doi":"10.21203/rs.3.rs-5538058/v1","editorialEvents":[],"status":"published","journal":{"display":false,"email":"[email protected]","identity":"nature","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"nature","sideBox":"Learn more about [Nature](http://www.nature.com/nature/)","snPcode":"","submissionUrl":"","title":"Nature","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"d11cd259-9073-4967-8aad-ecb208dfc327","owner":[],"postedDate":"December 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":41406816,"name":"Biological sciences/Chemical biology/Pharmacology/Receptor pharmacology"},{"id":41406817,"name":"Biological sciences/Drug discovery/Pharmacology/Receptor pharmacology"}],"tags":[],"updatedAt":"2025-10-23T07:10:53+00:00","versionOfRecord":{"articleIdentity":"rs-5538058","link":"https://doi.org/10.1038/s41586-025-09643-2","journal":{"identity":"nature","isVorOnly":false,"title":"Nature"},"publishedOn":"2025-10-22 04:00:00","publishedOnDateReadable":"October 22nd, 2025"},"versionCreatedAt":"2024-12-11 06:59:30","video":"","vorDoi":"10.1038/s41586-025-09643-2","vorDoiUrl":"https://doi.org/10.1038/s41586-025-09643-2","workflowStages":[]},"version":"v1","identity":"rs-5538058","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5538058","identity":"rs-5538058","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}