Functional Investigation of RNA Viral Helicases as Molecular Targets for the Discovery of Novel Antiviral Agents
type: CreativeWork
description: The recent SARS-CoV-2 pandemic highlighted the lack in pandemics preparedness for hCoVs potential future spillovers and for other (+)ssRNA viruses that still lack approved pharmacological treatments such as Flaviviruses. Small molecule development stands out as the most promising approach, as it enables possibly oral administration, stockpiling, combinatorial therapy. (+)ssRNA helicases, essential for viral replication, are among the most conserved proteins across several viral families, are a molecular class still underexplored, that lacks approved antiviral agents. In this respect, the identification of first-in-class inhibitor scaffolds is a promising achievement to pursue. The work is focused on the helicases of Coronaviruses, specifically the NSP13 of SARS-CoV-2, and of Flaviviruses, particularly the NS3 of WNV. Despite belonging to SF2 and SF1, respectively, the two proteins have strong similarities: both are non-ring forming helicases with core RecA-like domains; they share conserved motifs across the helicases and possess similar NTPase, helicase, and RTPase activities. SARS-CoV-2 NSP13 and WNV NS3helbiochemical assays and biophysical methods were established. The screening of chemically diverse compound identified first-in-class inhibitor scaffolds ranking among the strongest NSP13 and NS3 helicase inhibitors reported to date, based on IC50 and EC50 values and SI. Among a first set of 12 diketo acid derivatives, that inhibited both SARS-CoV-2 NSP13 enzymatic functions, three hit compounds, 5a,b,d, exhibited broad-spectrum activity against SARS-CoV-2, MERS-CoV, hCoV-OC43, and hCoV-229E. Mode of action studiesn suggested an ATP non-competitive inhibitory mechanism, likely targeting an allosteric pocket on the RecA2 domain. A rational design targeting the ATP-binding site region of SARS-CoV-2 NSP13 identified the 2-iminothiazolidin-4-one scaffold as promising core for antiviral molecules. Hit compound, 3i, potently inhibitNSP13 enzymatic functions, with a mixe…
abstract: The recent SARS-CoV-2 pandemic highlighted the lack in pandemics preparedness for hCoVs potential future spillovers and for other (+)ssRNA viruses that still lack approved pharmacological treatments such as Flaviviruses. Small molecule development stands out as the most promising approach, as it enables possibly oral administration, stockpiling, combinatorial therapy. (+)ssRNA helicases, essential for viral replication, are among the most conserved proteins across several viral families, are a molecular class still underexplored, that lacks approved antiviral agents. In this respect, the identification of first-in-class inhibitor scaffolds is a promising achievement to pursue. The work is focused on the helicases of Coronaviruses, specifically the NSP13 of SARS-CoV-2, and of Flaviviruses, particularly the NS3 of WNV. Despite belonging to SF2 and SF1, respectively, the two proteins have strong similarities: both are non-ring forming helicases with core RecA-like domains; they share conserved motifs across the helicases and possess similar NTPase, helicase, and RTPase activities. SARS-CoV-2 NSP13 and WNV NS3helbiochemical assays and biophysical methods were established. The screening of chemically diverse compound identified first-in-class inhibitor scaffolds ranking among the strongest NSP13 and NS3 helicase inhibitors reported to date, based on IC50 and EC50 values and SI. Among a first set of 12 diketo acid derivatives, that inhibited both SARS-CoV-2 NSP13 enzymatic functions, three hit compounds, 5a,b,d, exhibited broad-spectrum activity against SARS-CoV-2, MERS-CoV, hCoV-OC43, and hCoV-229E. Mode of action studiesn suggested an ATP non-competitive inhibitory mechanism, likely targeting an allosteric pocket on the RecA2 domain. A rational design targeting the ATP-binding site region of SARS-CoV-2 NSP13 identified the 2-iminothiazolidin-4-one scaffold as promising core for antiviral molecules. Hit compound, 3i, potently inhibitNSP13 enzymatic functions, with a mixe…
author: EMMOLO, ROBERTA
language: eng
keywords: Settore BIOS-15/A - Microbiologia
date published: 2026-07-13T00:00:00+02:00
date modified: 2026-07-24T01:29:55Z
provider: Università di Cagliari
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Università degli Studi di Cagliari

PropertyValue
property ID: Handle
value: https://hdl.handle.net/11584/489345