{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T20:59:54Z","timestamp":1790888394721,"version":"4.1.0"},"reference-count":63,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T00:00:00Z","timestamp":1722470400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University of Arizona R Ken Coit College of Pharmacy faculty startup fund","award":["AI175435"],"award-info":[{"award-number":["AI175435"]}]},{"name":"University of Arizona R Ken Coit College of Pharmacy faculty startup fund","award":["AI161845"],"award-info":[{"award-number":["AI161845"]}]},{"name":"University of Arizona R Ken Coit College of Pharmacy faculty startup fund","award":["AI131669"],"award-info":[{"award-number":["AI131669"]}]},{"name":"University of Arizona R Ken Coit College of Pharmacy faculty startup fund","award":["AI140726"],"award-info":[{"award-number":["AI140726"]}]},{"name":"R. Ken and Donna Coit Endowed Chair fund","award":["AI175435"],"award-info":[{"award-number":["AI175435"]}]},{"name":"R. Ken and Donna Coit Endowed Chair fund","award":["AI161845"],"award-info":[{"award-number":["AI161845"]}]},{"name":"R. Ken and Donna Coit Endowed Chair fund","award":["AI131669"],"award-info":[{"award-number":["AI131669"]}]},{"name":"R. Ken and Donna Coit Endowed Chair fund","award":["AI140726"],"award-info":[{"award-number":["AI140726"]}]},{"name":"NIH","award":["AI175435"],"award-info":[{"award-number":["AI175435"]}]},{"name":"NIH","award":["AI161845"],"award-info":[{"award-number":["AI161845"]}]},{"name":"NIH","award":["AI131669"],"award-info":[{"award-number":["AI131669"]}]},{"name":"NIH","award":["AI140726"],"award-info":[{"award-number":["AI140726"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Emergence of newer variants of SARS-CoV-2 underscores the need for effective antivirals to complement the vaccination program in managing COVID-19. The multi-functional papain-like protease (PLpro) of SARS-CoV-2 is an essential viral protein that not only regulates the viral replication but also modulates the host immune system, making it a promising therapeutic target. To this end, we developed an in vitro interferon stimulating gene 15 (ISG15)-based F\u00f6rster resonance energy transfer (FRET) assay and screened the National Cancer Institute (NCI) Diversity Set VI compound library, which comprises 1584 small molecules. Subsequently, we assessed the PLpro enzymatic activity in the presence of screened molecules. We identified three potential PLpro inhibitors, namely, NSC338106, 651084, and 679525, with IC50 values in the range from 3.3 to 6.0 \u00b5M. These molecules demonstrated in vitro inhibition of the enzyme activity and exhibited antiviral activity against SARS-CoV-2, with EC50 values ranging from 0.4 to 4.6 \u00b5M. The molecular docking of all three small molecules to PLpro suggested their specificity towards the enzyme\u2019s active site. Overall, our study contributes promising prospects for further developing potential antivirals to combat SARS-CoV-2 infection.<\/jats:p>","DOI":"10.3390\/v16081239","type":"journal-article","created":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T15:26:53Z","timestamp":1722526013000},"page":"1239","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["An ISG15-Based High-Throughput Screening Assay for Identification and Characterization of SARS-CoV-2 Inhibitors Targeting Papain-like Protease"],"prefix":"10.3390","volume":"16","author":[{"given":"Subodh Kumar","family":"Samrat","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prashant","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchen","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2570-8120","authenticated-orcid":false,"given":"Hyun","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy and Biophysics Core, Research Resources Center, University of Illinois at Chicago, Chicago, IL 60607, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yin","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongmin","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ 85721, USA"},{"name":"Department of Chemistry and Biochemistry, College of Science & College of Medicine, The University of Arizona, Tucson, AZ 85721, USA"},{"name":"The BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/s41577-021-00662-4","article-title":"SARS-CoV-2 breakthrough infections in vaccinated individuals: Measurement, causes and impact","volume":"22","author":"Lipsitch","year":"2022","journal-title":"Nat. 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