{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T19:11:29Z","timestamp":1754161889837,"version":"3.41.2"},"reference-count":31,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T00:00:00Z","timestamp":1752710400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001552","name":"Cancer Research Trust New Zealand","doi-asserted-by":"publisher","award":["GOT\u20101550\u2010RPG"],"award-info":[{"award-number":["GOT\u20101550\u2010RPG"]}],"id":[{"id":"10.13039\/501100001552","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Aberrant human growth hormone (GH) expression has been increasingly linked to poor outcomes in certain cancers and reduced health span. GH receptor (GHR) signal transduction activates signaling pathways that promote cell proliferation and survival, contributing to tumor progression. Consequently, there has been growing interest in creating antagonists that target GH signaling for anticancer applications. Pegvisomant is the only clinically available GHR antagonist (GHA). It is a protein antagonist that consists of a mutated GH protein (B2036) conjugated to polyethylene glycol (PEG). While pegvisomant is a potent human GHR antagonist, it has lower efficacy in mice, necessitating the use of higher doses in preclinical models. Here, we describe two antagonists, GHA2 and GHA3, based on an earlier reported antagonist, B2024, that have improved efficacy in mice. Recombinant GHA2 and GHA3 were produced in <jats:italic>E. coli<\/jats:italic> and then conjugated to multiple 5\u2009kDa amine\u2010reactive mPEG (GHA2\u2010PEG5K) or site\u2010specifically PEGylated with 40\u2009kDa mPEG at amino acid site 144 (GHA3\u2010PEG40K). When compared with B2036, GHA2 and GHA3 had significantly improved in vitro activity against the mouse GHR in a Ba\/F3 cell viability assay, and GHA2 more effectively reduced GH\u2010dependent signal transduction in the mouse melanoma cell line, B16\u2010F10. A single dose of 30\u2009mg\/kg of amine\u2010PEGylated GHA2\u2010PEG5K or site\u2010specific conjugate, GHA3\u2010PEG40K, reduced serum IGF\u20101 in mice at 24\u2009h by 40.7% and 45.8%, respectively. In contrast, amine\u2010PEGylated B2036\u2010PEG5K did not significantly reduce serum IGF\u2010I levels. In conclusion, we demonstrate here that GHA2\u2010PEG5K and GHA3\u2010PEG40K are more potent inhibitors than B2036\u2010PEG5K in mouse models.<\/jats:p>","DOI":"10.1002\/pro.70211","type":"journal-article","created":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T07:51:27Z","timestamp":1752738687000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Growth hormone receptor antagonists, <scp>GHA2<\/scp> and <scp>GHA3<\/scp>, with dual activity against the human and mouse receptor"],"prefix":"10.1002","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9010-4177","authenticated-orcid":false,"given":"Yue","family":"Wang","sequence":"first","affiliation":[{"name":"Liggins Institute University of Auckland  Auckland New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ries J.","family":"Langley","sequence":"additional","affiliation":[{"name":"Maurice Wilkins Centre for Molecular Biodiscovery  Auckland New Zealand"},{"name":"Department of Molecular Medicine and Pathology University of Auckland  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia","family":"Harms","sequence":"additional","affiliation":[{"name":"Liggins Institute University of Auckland  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Theo","family":"Portlock","sequence":"additional","affiliation":[{"name":"Liggins Institute University of Auckland  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen M. F.","family":"Jamieson","sequence":"additional","affiliation":[{"name":"Maurice Wilkins Centre for Molecular Biodiscovery  Auckland New Zealand"},{"name":"Auckland Cancer Society Research Centre University of Auckland  Auckland New Zealand"},{"name":"Department of Pharmacology and Clinical Pharmacology University of Auckland.  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4418-947X","authenticated-orcid":false,"given":"Jo K.","family":"Perry","sequence":"additional","affiliation":[{"name":"Liggins Institute University of Auckland  Auckland New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery  Auckland New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,7,17]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1210\/endrev\/bnae030"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.15375"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1249783"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)40764-2"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2020.111075"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.36.21969"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-019-0071-6"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1210\/me.2008-0128"},{"key":"e_1_2_9_10_1","unstructured":"CunninghamBC BrunoS WellsJA ClarkRG OlsonK FuhGG.USOO6136563A 11 Patent Number: 6.2000."},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1948064"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1549776"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ygcen.2014.11.004"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1039\/C8CS00606G"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1210\/er.2001-0022"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1210\/me.2015-1319"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1993.1612"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/jps.21152"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2002-022049"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M303985200"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajmg.c.31357"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ghir.2006.09.006"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1097\/MED.0b013e328363183a"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tem.2017.05.003"},{"key":"e_1_2_9_25_1","unstructured":"Pharmacia & Upjohn.21\u2010106 Somavert Approval \u2010 FDA.2003.Https:\/\/Www.Accessdata.Fda.Gov\/Drugsatfda_docs\/Nda\/2003\/21\u2013106_Somavert_Pharmr_P2.Pdf"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.beem.2008.08.011"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1530\/ERC-23-0033"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1530\/JME-25-0007"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biomac.0c01105"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.bioconjchem.0c00208"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20141293"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1210\/me.2010-0255"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.70211","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T13:57:32Z","timestamp":1753797452000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.70211"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,17]]},"references-count":31,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2025,8]]}},"alternative-id":["10.1002\/pro.70211"],"URL":"https:\/\/doi.org\/10.1002\/pro.70211","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,17]]},"assertion":[{"value":"2024-12-17","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-06-16","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-17","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70211"}}