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More than 20% of the US population suffers from LPR and there is no effective medical therapy. Pepsin is a predominant source of damage during LPR which disrupts laryngeal barrier function potentially via E\u2010cadherin cleavage proteolysis and downstream matrix metalloproteinase (MMP) dysregulation. Fosamprenavir (FDA\u2010approved HIV therapeutic and prodrug of amprenavir) is a pepsin\u2010inhibiting LPR therapeutic candidate shown to rescue damage in an LPR mouse model. This study aimed to examine amprenavir protection against laryngeal monolayer disruption and related E\u2010cadherin proteolysis and MMP dysregulation in vitro.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Laryngeal (TVC HPV) cells were exposed to buffered saline, pH\u20097.4 or pH\u20094\u2009\u00b1\u20091\u2009mg\/mL pepsin\u2009\u00b1\u2009amprenavir (10\u201360\u2009min). Analysis was performed by microscopy, Western blot, and real time polymerase chain reaction (qPCR).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Amprenavir (1\u2009\u03bcM) rescued pepsin acid\u2010mediated cell dissociation (<jats:italic>p<\/jats:italic>\u2009&lt;\u2009.05). Pepsin acid caused E\u2010cadherin cleavage indicative of regulated intramembrane proteolysis (RIP) and increased <jats:italic>MMP\u20101,3,7,9,14<\/jats:italic> 24\u2010h postexposure (<jats:italic>p<\/jats:italic>\u2009&lt;\u2009.05). Acid alone did not cause cell dissociation or E\u2010cadherin cleavage. Amprenavir (10\u2009\u03bcM) protected against E\u2010cadherin cleavage and <jats:italic>MMP\u20101,9,14<\/jats:italic> induction (<jats:italic>p<\/jats:italic>\u2009&lt;\u2009.05).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Amprenavir, at serum concentrations achievable provided the manufacturer's recommended dose of fosamprenavir for HIV, protects against pepsin\u2010mediated cell dissociation, E\u2010cadherin cleavage, and MMP dysregulation thought to contribute to barrier dysfunction and related symptoms during LPR. Fosamprenavir to amprenavir conversion by laryngeal epithelia, serum and saliva, and relative drug efficacies in an LPR mouse model are under investigation to inform development of inhaled formulations for LPR.<\/jats:p><\/jats:sec>","DOI":"10.1002\/lio2.1102","type":"journal-article","created":{"date-parts":[[2023,6,24]],"date-time":"2023-06-24T18:04:24Z","timestamp":1687629864000},"page":"953-962","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Amprenavir inhibits pepsin\u2010mediated laryngeal epithelial disruption and E\u2010cadherin cleavage in vitro"],"prefix":"10.1002","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7045-357X","authenticated-orcid":false,"given":"Tina L.","family":"Samuels","sequence":"first","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin  Milwaukee Wisconsin 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