Interference of small compounds and Mg2+ with dsRNA-binding fluorophores compromises the identification of SARS-CoV-2 RdRp inhibitors DOI Creative Commons
Susana Llanos, Bruno Di Geronimo,

Ester Casajús

и другие.

Scientific Reports, Год журнала: 2024, Номер 14(1)

Опубликована: Ноя. 15, 2024

The COVID-19 pandemic highlighted the need for rapid development of antiviral therapies. Viral RNA-dependent RNA polymerases (RdRp) are promising targets, and numerous virtual screenings potential inhibitors were conducted without validation identified hits. Here we have tested a set presumed RdRp in biochemical assays based on fluorometric detection activity or electrophoretic separation products. We find that is unreliable as screening method because many small compounds interfere with fluorophore binding to dsRNA, this effect enhanced by Mg2+ metal ions used nucleic acid polymerases. fact fluorimetric leads false-positive hits underscores requirement independent methods. also show suramin, one proposed could be validated biochemically, multi-polymerase inhibitor. While does not hinder its an agent, it cannot considered specific inhibitor SARS-CoV-2 RdRp.

Язык: Английский

Transformative Approaches in SARS-CoV-2 Management: Vaccines, Therapeutics and Future Direction DOI
Ankita Saha,

Shweta Choudhary,

Priyanshu Walia

и другие.

Virology, Год журнала: 2025, Номер 604, С. 110394 - 110394

Опубликована: Янв. 11, 2025

Язык: Английский

Процитировано

1
Structural basis of SARS-CoV-2 polymerase inhibition by nonnucleoside inhibitor HeE1-2Tyr DOI Creative Commons
Florian Kabinger,

Valerie Doze,

Jana Schmitzová

и другие.

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(10)

Опубликована: Март 4, 2025

Targeting the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with small molecules is a promising therapeutic strategy against COVID-19, but potent and safe inhibitors are lacking. HeE1-2Tyr, nonnucleoside inhibitor Dengue virus RdRp, was also shown to inhibit RdRp in vitro have antiviral activity cells, underlying mechanism remains unclear. Here, we elucidate molecular HeE1-2Tyr-mediated inhibition. Biochemical assays confirm that HeE1-2Tyr inhibits an IC 50 5 µM show it competes binding vitro. Structural analysis using cryo-EM reveals stack three binds site RdRp. The identification conserved its intriguing inhibition stacked outcompete may facilitate further development pan-corona inhibitors.

Язык: Английский

Процитировано

0
Structural insights into the RNA binding inhibitors of the C-terminal domain of the SARS-CoV-2 nucleocapsid DOI

Preeti Dhaka,

Jai Krishna Mahto,

Ankur Singh

и другие.

Journal of Structural Biology, Год журнала: 2025, Номер unknown, С. 108197 - 108197

Опубликована: Март 1, 2025

Язык: Английский

Процитировано

0
Structural insights into the RNA binding inhibitors of the C-terminal domain of the SARS-CoV-2 nucleocapsid DOI Creative Commons

Preeti Dhaka,

Jai Krishna Mahto,

Ankur Singh

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 11, 2024

Abstract The SARS-CoV-2 nucleocapsid (N) protein is an essential structural element of the virion, playing a crucial role in enclosing viral genome into ribonucleoprotein (RNP) assembly, as well replication and transmission. C-terminal domain N-protein (N-CTD) for encapsidation, contributing to stabilization RNP complex. In previous study, three inhibitors (ceftriaxone, cefuroxime, ampicillin) were screened their potential disrupt RNA packaging process by targeting N-protein. However, binding efficacy, mechanism inhibition, molecular insights with N-CTD remain unclear. this we evaluated efficacy these using isothermal titration calorimetry (ITC), revealing affinity ceftriaxone (18 ± 1.3 μM), cefuroxime (55 4.2 ampicillin (28 1.2 μM) N-CTD. Further inhibition assay fluorescence polarisation demonstrated IC 50 ranging from 10.4 12.4 μM K D values between 24 32 inhibitors. Additionally, also determined inhibitor-bound complex crystal structures N-CTD-Ceftriaxone (2.0 Å) N-CTD-Ampicillin (2.2 Å), along structure apo (1.4 Å). These revealed previously unobserved interaction sites involving residues K261, K266, R293, Q294, W301 at oligomerization interface predicted RNA-binding region findings provide valuable N-CTD, highlighting its underexplored but promising target development novel antiviral agents against coronaviruses. Highlights ceftriaxone, ampicillin-demonstrated high-affinity protein, effectively disrupting formation N-CTD-RNA Complex distinct sites. Structures reveal how selected hinder virus.

Язык: Английский

Процитировано

2
Plasmonic Imaging of Multivalent NTD–Nucleic Acid Interactions for Broad-Spectrum Antiviral Drug Analysis DOI

Yancao Chen,

Shijie Sun,

Xixuan Liu

и другие.

Analytical Chemistry, Год журнала: 2024, Номер 96(23), С. 9551 - 9560

Опубликована: Май 24, 2024

The discovery and identification of broad-spectrum antiviral drugs are great significance for blocking the spread pathogenic viruses corresponding variants concern. Herein, we proposed a plasmonic imaging-based strategy assessing efficacy potential targeting N-terminal domain nucleocapsid protein (NTD) nucleic acid (NA) interactions. With NTD NA conjugated gold nanoparticles as core satellite nanoprobes, respectively, found that multivalent binding interactions could drive formation core–satellite nanostructures with enhanced scattering brightness due to coupling effect. assembly can be suppressed in presence NTD–NA interactions, allowing drug analysis by detecting dose-dependent changes imaging. By quantifying uncovered constructed weak displayed 500-fold enhancement affinity compared monovalent We demonstrated evaluating drug, PJ34, target IC50 24.35 14.64 μM SARS-CoV-2 SARS-CoV, respectively. Moreover, discovered ceftazidime holds candidate inhibit an 22.08 from molecular docking analysis. Finally, validated 5-benzyloxygramine, which induce abnormal dimerization proteins, is effective SARS-CoV-2, but not against SARS-CoV. All these demonstrations indicated robust used powerful evolutionarily conserved viral proteins.

Язык: Английский

Процитировано

1
COVID-19 Genomic Surveillance in Bangui (Central African Republic) Reveals a Landscape of Circulating Variants Linked to Validated Antiviral Targets of SARS-CoV-2 Proteome DOI Creative Commons
Ulrich Vickos, Marianna Camasta, Nicole Grandi

и другие.

Viruses, Год журнала: 2023, Номер 15(12), С. 2309 - 2309

Опубликована: Ноя. 24, 2023

Since its outbreak, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) spread rapidly, causing the Disease 19 (COVID-19) pandemic. Even with vaccines’ administration, virus continued to circulate due inequal access prevention and therapeutic measures in African countries. Information about COVID-19 Africa has been limited contradictory, thus regional studies are important. On this premise, we conducted a genomic surveillance study lineages circulating Bangui, Central Republic (CAR). We collected 2687 nasopharyngeal samples at four checkpoints Bangui from 22 July 2021. Fifty-three tested positive for SARS-CoV-2, viral genomes were sequenced look presence of different strains. performed phylogenetic analysis described lineage landscape SARS-CoV-2 CAR along 15 months pandemics during period, finding Delta variant as predominant Variant Concern (VoC). The deduced aminoacidic sequences structural non-structural genes determined compared reference reported isolates Africa. Despite number obtained, provides valuable information evolution level allows better understanding circulation CAR.

Язык: Английский

Процитировано

3
Development of Fusion-Based Assay as a Drug Screening Platform for Nipah Virus Utilizing Baculovirus Expression Vector System DOI Open Access

Indah Permata Sari,

Christopher Llynard Ortiz, Lee‐Wei Yang

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(16), С. 9102 - 9102

Опубликована: Авг. 22, 2024

Nipah virus (NiV) is known to be a highly pathogenic zoonotic virus, which included in the World Health Organization Research & Development Blueprint list of priority diseases with up 70% mortality rate. Due its high pathogenicity and outbreak potency, therapeutic countermeasure against NiV urgently needed. As needs handled within Biological Safety Level (BSL) 4 facility, we had developed safe drug screening platform utilizing baculovirus expression vector system (BEVS) based on NiV-induced syncytium formation that could BSL-1 facility. To reconstruct BEVS, two baculoviruses were generated express recombinant proteins are responsible for inducing formation, including one exhibiting co-expressed fusion protein (NiV-F) attachment glycoprotein (NiV-G) another human EphrinB2 protein. Interestingly, was observed infected insect cells when medium modified have lower pH level supplemented cholesterol. Fusion inhibitory properties several compounds, such as phytochemicals polysulfonated naphthylamine compound, evaluated using this platform. Among these suramin showed highest activity system. Moreover, our silico results provide molecular-level glimpse suramin’s interaction NiV-G’s central hole EphrinB2’s G-H loop, possible reason activity.

Язык: Английский

Процитировано

0
The ALS drug riluzole binds to the C-terminal domain of SARS-CoV-2 nucleocapsid protein and has antiviral activity DOI
M.A. Marquez-Monino, Clara M. Santiveri, Patricia de León

и другие.

Structure, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 1, 2024

Язык: Английский

Процитировано

0
Interference of small compounds and Mg2+ with dsRNA-binding fluorophores compromises the identification of SARS-CoV-2 RdRp inhibitors DOI Creative Commons
Susana Llanos, Bruno Di Geronimo,

Ester Casajús

и другие.

Scientific Reports, Год журнала: 2024, Номер 14(1)

Опубликована: Ноя. 15, 2024

The COVID-19 pandemic highlighted the need for rapid development of antiviral therapies. Viral RNA-dependent RNA polymerases (RdRp) are promising targets, and numerous virtual screenings potential inhibitors were conducted without validation identified hits. Here we have tested a set presumed RdRp in biochemical assays based on fluorometric detection activity or electrophoretic separation products. We find that is unreliable as screening method because many small compounds interfere with fluorophore binding to dsRNA, this effect enhanced by Mg2+ metal ions used nucleic acid polymerases. fact fluorimetric leads false-positive hits underscores requirement independent methods. also show suramin, one proposed could be validated biochemically, multi-polymerase inhibitor. While does not hinder its an agent, it cannot considered specific inhibitor SARS-CoV-2 RdRp.

Язык: Английский

Процитировано

0