{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T12:51:20Z","timestamp":1791895880298,"version":"4.3.4"},"reference-count":62,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Glucagon is a well\u2010established therapeutic peptide, widely used to treat hypoglycemia. Like many peptide drugs, it offers advantages such as specificity, biocompatibility, and high affinity for receptor targets, but suffers from limited physical and chemical stability. In particular, improper conditions can promote the formation of amyloid fibrils, leading to loss of biological activity and, in some cases, cytotoxicity. Understanding the conditions that modulate the structural behavior of glucagon is therefore crucial. Currently, two injectable formulations are available on the market: a lyophilized vial of glucagon with lactose at acidic pH, and a more recent auto\u2010injector ready\u2010to\u2010use (RTU) formulation containing glucagon in dimethyl sulfoxide (DMSO) with trehalose. In this study, we investigated the conformational properties of glucagon in these two marketed formulations and compared them with glucagon dissolved in aqueous solution at pH\u20093.5, a metastable condition prone to aggregation. Preliminary circular dichroism and fluorescence spectroscopy were used to confirm glucagon stability over time; subsequently, NMR analysis showed that structural destabilization consistently begins at the C\u2010terminal region, while the\n                    <jats:sup>11<\/jats:sup>\n                    Ser\u2010Leu\n                    <jats:sup>14<\/jats:sup>\n                    segment remains structured across all environments. These findings highlight two key determinants of glucagon stability and aggregation. Strategies that preserve the integrity of the C\u2010terminal region while stabilizing the\n                    <jats:sup>11<\/jats:sup>\n                    Ser\u2010Leu\n                    <jats:sup>14<\/jats:sup>\n                    motif may improve peptide solubility and extend shelf life, providing a rational basis for the design of next\u2010generation glucagon formulations for emergency use.\n                  <\/jats:p>","DOI":"10.1002\/pro.70750","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T05:10:48Z","timestamp":1785993048000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Conformational determinants of glucagon stability and aggregation kinetics in aqueous and commercial formulations"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9690-907X","authenticated-orcid":false,"given":"Angelo","family":"Santoro","sequence":"first","affiliation":[{"name":"Department of Pharmacy University of Salerno  Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Macis","sequence":"additional","affiliation":[{"name":"Fresenius Kabi iPSUM  Rovigo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1189-1729","authenticated-orcid":false,"given":"Michela","family":"Buonocore","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences University of Naples \u201cFederico II,\u201d Complesso Universitario Monte Sant'Angelo  Naples Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1086-5852","authenticated-orcid":false,"given":"Antonio","family":"Ricci","sequence":"additional","affiliation":[{"name":"Fresenius Kabi iPSUM  Rovigo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6814-8472","authenticated-orcid":false,"given":"Anna Maria","family":"D'Ursi","sequence":"additional","affiliation":[{"name":"Department of Pharmacy University of Salerno  Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/pharmaceutics14112533"},{"key":"e_1_2_9_3_1","article-title":"Can mini\u2010dose glucagon prevent fasting\u2010induced hypoglycemia and preserve fasting?","volume":"17","author":"Algeffari M","year":"2025","journal-title":"Cureus"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1515\/jbcpp-2025-0005"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi6025374"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2364(85)90018-6"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1969.tb00735.x"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/dom.12663"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(83)80143-0"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2022.e11568"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1751-1097.1973.tb06422.x"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2022.10.001"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00682a037"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.3390\/antiox13121585"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcdhc.2022.889248"},{"key":"e_1_2_9_16_1","volume-title":"Advanced inorganic chemistry","author":"Cotton FA","year":"1999"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-012-0820-7"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41594-019-0238-6"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-012-9615-8"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1968.tb19527.x"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(04)83012-X"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.3390\/pharmaceutics14030681"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-809-9:353"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S2213-8587(14)70226-8"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/dom.15883"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.16312"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1063\/1.438208"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00028.2023"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep09005"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpharm.2024.123785"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-0-387-46312-4"},{"key":"e_1_2_9_32_1","first-page":"682","volume-title":"Maestro","author":"Maestro S","year":"2020"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2023.1148603"},{"key":"e_1_2_9_34_1","unstructured":"MengesF.Spectragryph\u2010optical spectroscopy software. Version 2020 1; 2016\u20132017."},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bioeng.4.092801.094202"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1023\/B:PHAM.0000033016.36825.2c"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.oprd.4c00515"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-458X(199804)36:4<245::AID-OMR264>3.0.CO;2-J"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/0010-4655(95)00041-D"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1177\/193229681000400609"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.molpharmaceut.4c00038"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1314474"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-811114-7.00006-2"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2025.184420"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24020971"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2022.114400"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.3390\/antiox14010053"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature02264"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.2337\/ds19-0076"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom10081192"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-0278(96)00046-6"},{"key":"e_1_2_9_52_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.26994"},{"key":"e_1_2_9_53_1","volume-title":"Physiology, glucagon","author":"Venugopal SK","year":"2023"},{"key":"e_1_2_9_54_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(01)76183-8"},{"key":"e_1_2_9_55_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkh371"},{"key":"e_1_2_9_56_1","doi-asserted-by":"publisher","DOI":"10.1002\/bip.20853"},{"key":"e_1_2_9_57_1","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2025.1541794"},{"key":"e_1_2_9_58_1","doi-asserted-by":"publisher","DOI":"10.1051\/epn\/19861701011"},{"key":"e_1_2_9_59_1","doi-asserted-by":"publisher","DOI":"10.1039\/D1NJ02230J"},{"key":"e_1_2_9_60_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi026629r"},{"key":"e_1_2_9_61_1","doi-asserted-by":"publisher","DOI":"10.1098\/rsfs.2017.0030"},{"key":"e_1_2_9_62_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature25153"},{"key":"e_1_2_9_63_1","doi-asserted-by":"publisher","DOI":"10.1210\/endocr\/bqaf130"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70750","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/pro.70750","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70750","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T12:24:53Z","timestamp":1791894293000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.70750"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":62,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/pro.70750"],"URL":"https:\/\/doi.org\/10.1002\/pro.70750","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]},"assertion":[{"value":"2026-03-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-27","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-05","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70750"}}