{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T17:09:23Z","timestamp":1790356163867,"version":"4.1.0"},"reference-count":91,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2021,2,11]],"date-time":"2021-02-11T00:00:00Z","timestamp":1613001600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000050","name":"National Heart, Lung, and Blood Institute","doi-asserted-by":"publisher","award":["F31HL146178"],"award-info":[{"award-number":["F31HL146178"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Niclosamide (NIC) has demonstrated promising <jats:italic>in vitro<\/jats:italic> antiviral efficacy against SARS-CoV-2, the causative agent of the COVID-19 pandemic. Though NIC is already FDA-approved, administration of the currently available oral formulation results in systemic drug levels that are too low for the inhibition of SARS-CoV-2. We hypothesized that the co-formulation of NIC with an endogenous protein, human lysozyme (hLYS), could enable the direct aerosol delivery of the drug to the respiratory tract as an alternative to oral delivery, thereby effectively treating COVID-19 by targeting the primary site of SARS-CoV-2 acquisition and spread. To test this hypothesis, we engineered and optimized composite particles containing NIC and hLYS suitable for delivery to the upper and lower airways via dry powder inhaler, nebulizer, and nasal spray. The novel formulation demonstrates potent <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> activity against two coronavirus strains, MERS-CoV and SARS-CoV-2, and may offer protection against methicillin-resistance staphylococcus aureus pneumonia and inflammatory lung damage occurring secondary to SARS-CoV-2 infections. The suitability of the formulation for all stages of the disease and low-cost development approach will ensure rapid clinical development and wide-spread utilization.<\/jats:p>","DOI":"10.1371\/journal.pone.0246803","type":"journal-article","created":{"date-parts":[[2021,2,12]],"date-time":"2021-02-12T00:07:21Z","timestamp":1613088441000},"page":"e0246803","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":62,"title":["Development and evaluation of inhalable composite niclosamide-lysozyme particles: A broad-spectrum, patient-adaptable treatment for coronavirus infections and sequalae"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5456-5427","authenticated-orcid":true,"given":"Ashlee D.","family":"Brunaugh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyojong","family":"Seo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary","family":"Warnken","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sang Heui","family":"Seo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6582-5869","authenticated-orcid":true,"given":"Hugh D. C.","family":"Smyth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,2,11]]},"reference":[{"key":"pone.0246803.ref001","unstructured":"WHO Director-General\u2019s opening remarks at the Mission briefing on COVID-19\u201312 March 2020 [Internet].; 2020"},{"issue":"5","key":"pone.0246803.ref002","doi-asserted-by":"crossref","first-page":"e208297","DOI":"10.1001\/jamanetworkopen.2020.8297","article-title":"Estimated Demand for US Hospital Inpatient and Intensive Care Unit Beds for Patients With COVID-19 Based on Comparisons With Wuhan and Guangzhou, China.","volume":"3","author":"R Li","year":"2020","journal-title":"JAMA Netw Open."},{"issue":"8","key":"pone.0246803.ref003","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1038\/s41591-020-0952-y","article-title":"Disease and healthcare burden of COVID-19 in the United States","volume":"26","author":"IF Miller","year":"2020","journal-title":"Nat Med"},{"issue":"6493","key":"pone.0246803.ref004","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1126\/science.abb5793","article-title":"Projecting the transmission dynamics of SARS-CoV-2 through the postpandemic period","volume":"368","author":"SM Kissler","year":"2020","journal-title":"Science"},{"issue":"7","key":"pone.0246803.ref005","doi-asserted-by":"crossref","first-page":"e00819","DOI":"10.1128\/AAC.00819-20","article-title":"Identification of Antiviral Drug Candidates against SARS-CoV-2 from FDA-Approved Drugs","volume":"64","author":"S Jeon","year":"2020","journal-title":"Antimicrobial Agents and Chemotherapy"},{"issue":"1","key":"pone.0246803.ref006","doi-asserted-by":"crossref","first-page":"5770","DOI":"10.1038\/s41467-019-13659-4","article-title":"SKP2 attenuates autophagy through Beclin1-ubiquitination and its inhibition reduces MERS-Coronavirus infection.","volume":"10","author":"NC Gassen","year":"2019","journal-title":"Nat Commun."},{"issue":"10","key":"pone.0246803.ref007","doi-asserted-by":"crossref","first-page":"e1002976","DOI":"10.1371\/journal.ppat.1002976","article-title":"Niclosamide is a proton carrier and targets acidic endosomes with broad antiviral effects.","volume":"8","author":"A Jurgeit","year":"2012","journal-title":"PLoS Pathog."},{"key":"pone.0246803.ref008","article-title":"Broad Spectrum Antiviral Agent Niclosamide and Its Therapeutic Potential.","author":"J Xu","year":"2020","journal-title":"ACS Infect Dis."},{"issue":"1","key":"pone.0246803.ref009","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.canlet.2014.04.003","article-title":"Multi-targeted therapy of cancer by niclosamide: A new application for an old drug","volume":"349","author":"Y Li","year":"2014","journal-title":"Cancer Lett"},{"key":"pone.0246803.ref010","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.cellsig.2017.04.001","article-title":"Niclosamide: Beyond an antihelminthic drug","volume":"41","author":"W Chen","year":"2018","journal-title":"Cell Signal"},{"issue":"1","key":"pone.0246803.ref011","doi-asserted-by":"crossref","first-page":"5233","DOI":"10.1038\/s41467-018-07705-w","article-title":"Host-targeted niclosamide inhibits C. difficile virulence and prevents disease in mice without disrupting the gut microbiota","volume":"9","author":"J Tam","year":"2018","journal-title":"Nat Commun"},{"issue":"21","key":"pone.0246803.ref012","doi-asserted-by":"crossref","first-page":"17530","DOI":"10.1074\/jbc.M112.359638","article-title":"Structure-activity analysis of niclosamide reveals potential role for cytoplasmic pH in control of mammalian target of rapamycin complex 1 (mTORC1) signaling.","volume":"287","author":"BD Fonseca","year":"2012","journal-title":"J Biol Chem"},{"issue":"1","key":"pone.0246803.ref013","doi-asserted-by":"crossref","first-page":"e0146931","DOI":"10.1371\/journal.pone.0146931","article-title":"A Novel High Content Imaging-Based Screen Identifies the Anti-Helminthic Niclosamide as an Inhibitor of Lysosome Anterograde Trafficking and Prostate Cancer Cell Invasion.","volume":"11","author":"ML Circu","year":"2016","journal-title":"PLoS One"},{"issue":"7","key":"pone.0246803.ref014","doi-asserted-by":"crossref","first-page":"e0159675","DOI":"10.1371\/journal.pone.0159675","article-title":"Extracellular pH Modulates Neuroendocrine Prostate Cancer Cell Metabolism and Susceptibility to the Mitochondrial Inhibitor Niclosamide.","volume":"11","author":"JE Ippolito","year":"2016","journal-title":"PLoS One"},{"issue":"17","key":"pone.0246803.ref015","doi-asserted-by":"crossref","first-page":"5829","DOI":"10.1016\/j.bmc.2015.07.001","article-title":"Structure-activity studies of Wnt\/beta-catenin inhibition in the Niclosamide chemotype: Identification of derivatives with improved drug exposure","volume":"23","author":"RA Mook","year":"2015","journal-title":"Bioorg Med Chem"},{"issue":"1","key":"pone.0246803.ref016","doi-asserted-by":"crossref","first-page":"3701","DOI":"10.1038\/s41598-018-22037-x","article-title":"Repurposing the anthelmintic drug niclosamide to combat Helicobacter pylori","volume":"8","author":"N Tharmalingam","year":"2018","journal-title":"Sci Rep"},{"issue":"4","key":"pone.0246803.ref017","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0124595","article-title":"Repurposing salicylanilide anthelmintic drugs to combat drug resistant Staphylococcus aureus.","volume":"10","author":"R Rajamuthiah","year":"2015","journal-title":"PloS one."},{"issue":"4","key":"pone.0246803.ref018","doi-asserted-by":"crossref","first-page":"045010","DOI":"10.1088\/1748-605X\/aa7105","article-title":"Repurposing niclosamide as a versatile antimicrobial surface coating against device-associated, hospital-acquired bacterial infections.","volume":"12","author":"T Gwisai","year":"2017","journal-title":"Biomedical Materials"},{"issue":"10","key":"pone.0246803.ref019","doi-asserted-by":"crossref","first-page":"5663","DOI":"10.1128\/AAC.00377-16","article-title":"Screening a commercial library of pharmacologically active small molecules against Staphylococcus aureus biofilms","volume":"60","author":"NS Torres","year":"2016","journal-title":"Antimicrobial agents and chemotherapy"},{"issue":"4","key":"pone.0246803.ref020","doi-asserted-by":"crossref","first-page":"e02574","DOI":"10.1128\/AAC.02574-18","article-title":"The Anthelmintic Drug Niclosamide Synergizes with Colistin and Reverses Colistin Resistance in Gram-Negative Bacilli","volume":"63","author":"R Domalaon","year":"2019","journal-title":"Antimicrobial Agents and Chemotherapy"},{"issue":"8","key":"pone.0246803.ref021","doi-asserted-by":"crossref","first-page":"2604","DOI":"10.1021\/acs.molpharmaceut.5b00098","article-title":"Toward repositioning niclosamide for antivirulence therapy of Pseudomonas aeruginosa lung infections: development of inhalable formulations through nanosuspension technology","volume":"12","author":"G Costabile","year":"2015","journal-title":"Molecular pharmaceutics"},{"issue":"2","key":"pone.0246803.ref022","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1128\/AAC.01952-12","article-title":"New life for an old drug: the anthelmintic drug niclosamide inhibits Pseudomonas aeruginosa quorum sensing","volume":"57","author":"F Imperi","year":"2013","journal-title":"Antimicrobial agents and chemotherapy"},{"issue":"348","key":"pone.0246803.ref023","article-title":"Synergistic Activity of Niclosamide in Combination With Colistin Against Colistin-Susceptible and Colistin-Resistant Acinetobacter baumannii and Klebsiella pneumoniae","volume":"8","author":"R Ayerbe-Algaba","year":"2018","journal-title":"Frontiers in Cellular and Infection Microbiology"},{"issue":"5","key":"pone.0246803.ref024","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1054\/tuld.1999.0212","article-title":"Antituberculosis activity of certain antifungal and antihelmintic drugs","volume":"79","author":"Z Sun","year":"1999","journal-title":"Tubercle and Lung disease"},{"issue":"3","key":"pone.0246803.ref025","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.1128\/AAC.02154-12","article-title":"Activities of Drug Combinations against <span class = \"named-content genus-species\" id = \"named-content-1\">Mycobacterium tuberculosis Grown in Aerobic and Hypoxic Acidic Conditions.","volume":"57","author":"G Piccaro","year":"2013","journal-title":"Antimicrobial agents and chemotherapy"},{"issue":"4","key":"pone.0246803.ref026","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1134\/S0026261717040154","article-title":"Niclosamide as a promising antibiofilm agent","volume":"86","author":"MV Zhurina","year":"2017","journal-title":"Microbiology"},{"issue":"15","key":"pone.0246803.ref027","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.128414","article-title":"Niclosamide repurposed for the treatment of inflammatory airway disease.","volume":"4","author":"I Cabrita","year":"2019","journal-title":"JCI Insight."},{"key":"pone.0246803.ref028","unstructured":"PubChem Database. Niclosamide, CID = 4477, [Internet]. [cited Apr. 28, 2020]. Available from: https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/Niclosamide."},{"key":"pone.0246803.ref029","first-page":"2020","article-title":"Identification of antiviral drug candidates against SARS-CoV-2 from FDA-approved drugs.","author":"S Jeon","year":"2020","journal-title":"bioRxiv"},{"issue":"6","key":"pone.0246803.ref030","doi-asserted-by":"crossref","first-page":"e0198389","DOI":"10.1371\/journal.pone.0198389","article-title":"A phase I study of niclosamide in combination with enzalutamide in men with castration-resistant prostate cancer","volume":"13","author":"MT Schweizer","year":"2018","journal-title":"PloS one"},{"issue":"8","key":"pone.0246803.ref031","doi-asserted-by":"crossref","first-page":"2604","DOI":"10.1021\/acs.molpharmaceut.5b00098","article-title":"Toward Repositioning Niclosamide for Antivirulence Therapy of Pseudomonas aeruginosa Lung Infections: Development of Inhalable Formulations through Nanosuspension Technology","volume":"12","author":"G Costabile","year":"2015","journal-title":"Mol Pharm"},{"key":"pone.0246803.ref032","unstructured":"Inactive Ingredients Database [Internet]. 2019. Available from: https:\/\/www.accessdata.fda.gov\/scripts\/cder\/iig\/index.cfm."},{"issue":"11","key":"pone.0246803.ref033","doi-asserted-by":"crossref","first-page":"1407","DOI":"10.4187\/respcare.06153","article-title":"Nebulized Mannitol, Particle Distribution, and Cough in Idiopathic Pulmonary Fibrosis.","volume":"63","author":"PM Kanth","year":"2018","journal-title":"Respir Care"},{"issue":"4","key":"pone.0246803.ref034","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1111\/j.1440-1843.2005.00724.x","article-title":"Dissociation in the effect of nedocromil on mannitol-induced cough or bronchoconstriction in asthmatic subjects*.","volume":"10","author":"HO KOSKELA","year":"2005","journal-title":"Respirology"},{"key":"pone.0246803.ref035","article-title":"Post-inhalation cough with therapeutic aerosols: Formulation considerations","author":"S Sahakijpijarn","year":"2020","journal-title":"Adv Drug Deliv Rev"},{"issue":"3","key":"pone.0246803.ref036","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.jmii.2020.02.012","article-title":"Asymptomatic carrier state, acute respiratory disease, and pneumonia due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2): Facts and myths.","volume":"53","author":"CC Lai","year":"2020","journal-title":"J Microbiol Immunol Infect"},{"issue":"5","key":"pone.0246803.ref037","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.1007\/s10753-015-0171-8","article-title":"Anti-Inflammatory Effects of Lysozyme Against HMGB1 in Human Endothelial Cells and in Mice","volume":"38","author":"W Lee","year":"2015","journal-title":"Inflammation"},{"issue":"4660","key":"pone.0246803.ref038","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1038\/183548a0","article-title":"Antiviral Activity of Lysozyme.","volume":"183","author":"R Ferrari","year":"1959","journal-title":"Nature"},{"issue":"1","key":"pone.0246803.ref039","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1186\/s12917-019-2067-6","article-title":"Antiviral effects of nisin, lysozyme, lactoferrin and their mixtures against bovine viral diarrhoea virus","volume":"15","author":"J Malaczewska","year":"2019","journal-title":"BMC Vet Res"},{"issue":"1","key":"pone.0246803.ref040","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/S1671-2927(08)60029-2","article-title":"Study on Antimicrobial and Antiviral Activities of Lysozyme From Marine Strain S-12-86 In Vitro.","volume":"7","author":"X Zhang","year":"2008","journal-title":"Agricultural Sciences in China"},{"issue":"12","key":"pone.0246803.ref041","doi-asserted-by":"crossref","first-page":"4648","DOI":"10.1021\/bi0477081","article-title":"Structural and Functional Modeling of Human Lysozyme Reveals a Unique Nonapeptide, HL9, with Anti-HIV Activity","volume":"44","author":"S Lee-Huang","year":"2005","journal-title":"Biochemistry"},{"issue":"18","key":"pone.0246803.ref042","doi-asserted-by":"crossref","first-page":"10336","DOI":"10.1021\/jf2020396","article-title":"Human Lysozyme Possesses Novel Antimicrobial Peptides within Its N-terminal Domain that Target Bacterial Respiration","volume":"59","author":"HR Ibrahim","year":"2011","journal-title":"Journal of Agricultural and Food Chemistry"},{"issue":"5","key":"pone.0246803.ref043","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1165\/ajrcmb.20.5.3572","article-title":"Activity of abundant antimicrobials of the human airway","volume":"20","author":"SM Travis","year":"1999","journal-title":"American journal of respiratory cell and molecular biology"},{"issue":"9","key":"pone.0246803.ref044","doi-asserted-by":"crossref","first-page":"e1006512","DOI":"10.1371\/journal.ppat.1006512","article-title":"From bacterial killing to immune modulation: Recent insights into the functions of lysozyme.","volume":"13","author":"SA Ragland","year":"2017","journal-title":"PLoS Pathog."},{"key":"pone.0246803.ref045","article-title":"Effect of Particle Formation Process on Characteristics and Aerosol Performance of Respirable Protein Powders","author":"AD Brunaugh","year":"2019","journal-title":"Molecular Pharmaceutics"},{"issue":"7","key":"pone.0246803.ref046","doi-asserted-by":"crossref","first-page":"2755","DOI":"10.1208\/s12249-018-0980-9","article-title":"Influence of Formulation Factors on the Aerosol Performance and Stability of Lysozyme Powders: a Systematic Approach.","volume":"19","author":"S Ferrati","year":"2018","journal-title":"AAPS PharmSciTech."},{"key":"pone.0246803.ref047","volume-title":"Aerosol technology: properties, behavior, and measurement of airborne particles","author":"WC Hinds","year":"1999"},{"key":"pone.0246803.ref048","unstructured":"Lenth RV, Lenth MRV, Vdgraph S. Package \u2018rsm\u2019. 2018."},{"key":"pone.0246803.ref049","volume-title":"R: A language and environment for statistical computing","author":"R Development Core Team","year":"2019"},{"issue":"5","key":"pone.0246803.ref050","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1007\/s11095-010-0070-5","article-title":"In vitro aqueous fluid-capacity-limited dissolution testing of respirable aerosol drug particles generated from inhaler products","volume":"27","author":"D Arora","year":"2010","journal-title":"Pharm Res"},{"issue":"8","key":"pone.0246803.ref051","doi-asserted-by":"crossref","first-page":"2618","DOI":"10.1021\/acs.molpharmaceut.5b00221","article-title":"Evaluation of the Transwell System for Characterization of Dissolution Behavior of Inhalation Drugs: Effects of Membrane and Surfactant","volume":"12","author":"M Rohrschneider","year":"2015","journal-title":"Mol Pharm"},{"key":"pone.0246803.ref052","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/j.jddst.2018.08.006","article-title":"Design and development of a biorelevant simulated human lung fluid","volume":"47","author":"M Hassoun","year":"2018","journal-title":"J Drug Deliv Sci Technol"},{"issue":"11","key":"pone.0246803.ref053","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1111\/j.2042-7158.2012.01523.x","article-title":"Overcoming sink limitations in dissolution testing: a review of traditional methods and the potential utility of biphasic systems","volume":"64","author":"DJ Phillips","year":"2012","journal-title":"Journal of Pharmacy and Pharmacology"},{"issue":"3","key":"pone.0246803.ref054","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1089\/089426803769017668","article-title":"Next generation pharmaceutical impactor (a new impactor for pharmaceutical inhaler testing). Part II: Archival calibration.","volume":"16","author":"VA Marple","year":"2003","journal-title":"Journal of Aerosol medicine"},{"key":"pone.0246803.ref055","article-title":"Personalized medicine in nasal delivery: the use of patient-specific administration parameters to improve nasal drug targeting using 3D printed nasal replica casts","author":"ZN Warnken","year":"2018","journal-title":"Molecular Pharmaceutics"},{"issue":"3","key":"pone.0246803.ref056","doi-asserted-by":"crossref","first-page":"359","DOI":"10.3109\/03639045.2010.520321","article-title":"Automated actuation of nasal spray products: determination and comparison of adult and pediatric settings","volume":"37","author":"DV Doughty","year":"2011","journal-title":"Drug Development and Industrial Pharmacy"},{"issue":"7","key":"pone.0246803.ref057","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image analysis","volume":"9","author":"J Schindelin","year":"2012","journal-title":"Nature methods"},{"issue":"3","key":"pone.0246803.ref058","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1039\/c0fo00113a","article-title":"Transcription profiles of LPS-stimulated THP-1 monocytes and macrophages: a tool to study inflammation modulating effects of food-derived compounds","volume":"1","author":"W Chanput","year":"2010","journal-title":"Food Funct"},{"issue":"6","key":"pone.0246803.ref059","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1007\/s10529-016-2079-2","article-title":"Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants","volume":"38","author":"M Elshikh","year":"2016","journal-title":"Biotechnol Lett"},{"issue":"2","key":"pone.0246803.ref060","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.cell.2020.05.042","article-title":"SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract","volume":"182","author":"YJ Hou","year":"2020","journal-title":"Cell"},{"issue":"1\u20132","key":"pone.0246803.ref061","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ijpharm.2012.04.040","article-title":"Immunogenicity of protein aggregates\u2014concerns and realities","volume":"431","author":"W Wang","year":"2012","journal-title":"Int J Pharm"},{"issue":"2","key":"pone.0246803.ref062","doi-asserted-by":"crossref","first-page":"99","DOI":"10.3109\/1547691X.2013.821564","article-title":"Immunogenicity of therapeutic proteins: influence of aggregation","volume":"11","author":"KD Ratanji","year":"2014","journal-title":"J Immunotoxicol"},{"issue":"2","key":"pone.0246803.ref063","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.xphs.2015.11.002","article-title":"Immunogenicity of Therapeutic Protein Aggregates","volume":"105","author":"EM Moussa","year":"2016","journal-title":"J Pharm Sci"},{"issue":"1\u20132","key":"pone.0246803.ref064","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/S0378-5173(02)00379-4","article-title":"Integrity of crystalline lysozyme exceeds that of a spray-dried form","volume":"247","author":"AA Elkordy","year":"2002","journal-title":"International journal of pharmaceutics"},{"issue":"1","key":"pone.0246803.ref065","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s12038-010-0015-5","article-title":"Lysozymes in the animal kingdom","volume":"35","author":"L Callewaert","year":"2010","journal-title":"J Biosci"},{"issue":"3","key":"pone.0246803.ref066","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1046\/j.1365-2796.2003.01228.x","article-title":"Antibacterial peptides: basic facts and emerging concepts","volume":"254","author":"HG Boman","year":"2003","journal-title":"Journal of Internal Medicine"},{"issue":"5","key":"pone.0246803.ref067","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/j.molcel.2018.07.009","article-title":"An Antiviral Branch of the IL-1 Signaling Pathway Restricts Immune-Evasive Virus Replication","volume":"71","author":"MH Orzalli","year":"2018","journal-title":"Mol Cell"},{"issue":"1","key":"pone.0246803.ref068","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/1742-4933-11-4","article-title":"Severe acute respiratory syndrome-coronavirus infection in aged nonhuman primates is associated with modulated pulmonary and systemic immune responses.","volume":"11","author":"CC Clay","year":"2014","journal-title":"Immunity & Ageing."},{"issue":"6","key":"pone.0246803.ref069","doi-asserted-by":"crossref","first-page":"2815","DOI":"10.4049\/jimmunol.1103051","article-title":"Impaired NLRP3 inflammasome function in elderly mice during influenza infection is rescued by treatment with nigericin","volume":"188","author":"HW Stout-Delgado","year":"2012","journal-title":"J Immunol"},{"issue":"7","key":"pone.0246803.ref070","doi-asserted-by":"crossref","DOI":"10.1039\/c2sc20045g","article-title":"Rethinking the term \u201cpi-stacking\u201d.","volume":"3","author":"CR Martinez","year":"2012","journal-title":"Chemical Science"},{"issue":"7","key":"pone.0246803.ref071","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1007\/s13361-019-02166-x","article-title":"An In Vitro Study of Aromatic Stacking of Drug Molecules","volume":"30","author":"SN Jackson","year":"2019","journal-title":"J Am Soc Mass Spectrom"},{"issue":"9","key":"pone.0246803.ref072","doi-asserted-by":"crossref","first-page":"4588","DOI":"10.1021\/cg300784v","article-title":"Pharmaceutical Cocrystals of Niclosamide.","volume":"12","author":"P Sanphui","year":"2012","journal-title":"Crystal Growth & Design."},{"issue":"45","key":"pone.0246803.ref073","doi-asserted-by":"crossref","first-page":"14382","DOI":"10.1021\/acs.jpcb.5b07277","article-title":"Complexation of Lysozyme with Sodium Poly(styrenesulfonate) via the Two-State and Non-Two-State Unfoldings of Lysozyme.","volume":"119","author":"FG Wu","year":"2015","journal-title":"J Phys Chem B"},{"key":"pone.0246803.ref074","doi-asserted-by":"crossref","first-page":"117623","DOI":"10.1016\/j.saa.2019.117623","article-title":"Investigation of complexation of amlodipine with lysozyme and its effect on lysozyme crystal growth","volume":"227","author":"S Pawar","year":"2020","journal-title":"Spectrochim Acta A Mol Biomol Spectrosc"},{"key":"pone.0246803.ref075","unstructured":"COVID-19 Update: FDA Broadens Emergency Use Authorization for Veklury (remdesivir) to Include All Hospitalized Patients for Treatment of COVID-19 [Internet]. 2020. Available from: https:\/\/www.fda.gov\/news-events\/press-announcements\/covid-19-update-fda-broadens-emergency-use-authorization-veklury-remdesivir-include-all-hospitalized"},{"key":"pone.0246803.ref076","article-title":"Remdesivir for the Treatment of Covid-19\u2014Preliminary Report","author":"JH Beigel","year":"2020","journal-title":"N Engl J Med"},{"key":"pone.0246803.ref077","unstructured":"Safety, Tolerability and Pharmacokinetics of Inhaled Nanoparticle Formulation of Remdesivir (GS-5734) and NA-831. https:\/\/ClinicalTrials.gov\/show\/NCT04480333."},{"key":"pone.0246803.ref078","unstructured":"Study to Evaluate the Safety and Antiviral Activity of Remdesivir (GS-5734\u2122) in Participants With Moderate Coronavirus Disease (COVID-19) Compared to Standard of Care Treatment. https:\/\/ClinicalTrials.gov\/show\/NCT04292730."},{"issue":"11","key":"pone.0246803.ref079","doi-asserted-by":"crossref","DOI":"10.3390\/pharmaceutics12111002","article-title":"Development of Remdesivir as a Dry Powder for Inhalation by Thin Film Freezing.","volume":"12","author":"S Sahakijpijarn","year":"2020","journal-title":"Pharmaceutics"},{"issue":"1","key":"pone.0246803.ref080","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/s41467-019-13940-6","article-title":"Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.","volume":"11","author":"TP Sheahan","year":"2020","journal-title":"Nat Commun."},{"key":"pone.0246803.ref081","article-title":"An inflammatory cytokine signature predicts COVID-19 severity and survival","author":"DM Del Valle","year":"2020","journal-title":"Nat Med"},{"issue":"10229","key":"pone.0246803.ref082","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1016\/S0140-6736(20)30566-3","article-title":"Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study","volume":"395","author":"F Zhou","year":"2020","journal-title":"The Lancet"},{"issue":"2","key":"pone.0246803.ref083","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/0163-7258(82)90064-X","article-title":"The biology and toxicology of molluscicides, bayluscide.","volume":"19","author":"P Andrews","year":"1982","journal-title":"Pharmacology & Therapeutics."},{"issue":"3","key":"pone.0246803.ref084","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1007\/s11095-016-2050-x","article-title":"The Impact of Inspiratory Flow Rate on Drug Delivery to the Lungs with Dry Powder Inhalers","volume":"34","author":"J Weers","year":"2017","journal-title":"Pharm Res"},{"key":"pone.0246803.ref085","article-title":"Dysregulation of immune response in patients with COVID-19 in Wuhan, China","author":"C Qin","year":"2020","journal-title":"Clin Infect Dis"},{"key":"pone.0246803.ref086","doi-asserted-by":"crossref","first-page":"181","DOI":"10.3389\/fphar.2016.00181","article-title":"Measurements of Deposition, Lung Surface Area and Lung Fluid for Simulation of Inhaled Compounds.","volume":"7","author":"E Frohlich","year":"2016","journal-title":"Front Pharmacol"},{"issue":"1\u20132","key":"pone.0246803.ref087","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/S0378-5173(03)00039-5","article-title":"Brain delivery of vasoactive intestinal peptide (VIP) following nasal administration to rats.","volume":"255","author":"C Dufes","year":"2003","journal-title":"International journal of pharmaceutics"},{"key":"pone.0246803.ref088","article-title":"Ciclesonide, a hypotonic intranasal corticosteroid","author":"H Neffen","year":"2010","journal-title":"Allergy & Asthma Proceedings"},{"key":"pone.0246803.ref089","article-title":"Neurological manifestations of hospitalized patients with COVID-19 in Wuhan, China: a retrospective case series study.","author":"L Mao","year":"2020"},{"key":"pone.0246803.ref090","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.bbi.2020.03.031","article-title":"Nervous system involvement after infection with COVID-19 and other coronaviruses","volume":"87","author":"Y Wu","year":"2020","journal-title":"Brain Behav Immun."},{"issue":"1","key":"pone.0246803.ref091","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41467-019-13940-6","article-title":"Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV","volume":"11","author":"TP Sheahan","year":"2020","journal-title":"Nature communications"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0246803","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,2,12]],"date-time":"2021-02-12T00:08:42Z","timestamp":1613088522000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0246803"}},"subtitle":[],"editor":[{"given":"Nitesh Kumar","family":"Kunda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2021,2,11]]},"references-count":91,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,2,11]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0246803","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,11]]}}}