DHFR Inhibitors Display a Pleiotropic Anti-Viral Activity against SARS-CoV-2: Insights into the Mechanisms of Action

Daniela Iaconis; Francesca Caccuri; Candida Manelfi; Carmine Talarico; Antonella Bugatti; Federica Filippini; Alberto Zani; Rubina Novelli; Maria Kuzikov; Bernhard Ellinger; Philip Gribbon; Kristoffer Riecken; Francesca Esposito; Angela Corona; Enzo Tramontano; Andrea Rosario Beccari; Arnaldo Caruso; Marcello Allegretti

doi:10.3390/v15051128

DHFR Inhibitors Display a Pleiotropic Anti-Viral Activity against SARS-CoV-2: Insights into the Mechanisms of Action

Viruses. 2023 May 9;15(5):1128. doi: 10.3390/v15051128.

Authors

Daniela Iaconis¹, Francesca Caccuri², Candida Manelfi¹, Carmine Talarico¹, Antonella Bugatti², Federica Filippini², Alberto Zani², Rubina Novelli³, Maria Kuzikov^{4

5}, Bernhard Ellinger^{4

5}, Philip Gribbon^{4

5}, Kristoffer Riecken⁶, Francesca Esposito⁷, Angela Corona⁷, Enzo Tramontano⁷, Andrea Rosario Beccari¹, Arnaldo Caruso², Marcello Allegretti³

Affiliations

¹ EXSCALATE, Dompé Farmaceutici SpA, Via Tommaso De Amicis, 95, 80131 Napoli, Italy.
² Section of Microbiology Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy.
³ Dompè Famaceutici SpA, Via Campo di Pile snc, 67100 L'Aquila, Italy.
⁴ Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Schnackenburgallee 114, 22525 Hamburg, Germany.
⁵ Fraunhofer Cluster of Excellence for Immune-Mediated Diseases CIMD, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany.
⁶ Research Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
⁷ Department of Life and Environmental Sciences, University of Cagliari, Cittadella Universitaria SS554, 09042 Monserrato (CA), Italy.

Abstract

During the COVID-19 pandemic, drug repurposing represented an effective strategy to obtain quick answers to medical emergencies. Based on previous data on methotrexate (MTX), we evaluated the anti-viral activity of several DHFR inhibitors in two cell lines. We observed that this class of compounds showed a significant influence on the virus-induced cytopathic effect (CPE) partly attributed to the intrinsic anti-metabolic activity of these drugs, but also to a specific anti-viral function. To elucidate the molecular mechanisms, we took advantage of our EXSCALATE platform for in-silico molecular modelling and further validated the influence of these inhibitors on nsp13 and viral entry. Interestingly, pralatrexate and trimetrexate showed superior effects in counteracting the viral infection compared to other DHFR inhibitors. Our results indicate that their higher activity is due to their polypharmacological and pleiotropic profile. These compounds can thus potentially give a clinical advantage in the management of SARS-CoV-2 infection in patients already treated with this class of drugs.

Keywords: COVID-19; EXSCALATE; SARS-CoV-2; anti-viral activity; drug repurposing; methotrexate; molecular docking; nsp13; viral entry; virtual screening.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antiviral Agents / metabolism
Antiviral Agents / pharmacology
COVID-19*
Drug Repositioning / methods
Humans
Molecular Docking Simulation
Pandemics
SARS-CoV-2* / metabolism

Substances

Antiviral Agents

Grants and funding

This research was conducted under the project “EXaSCale smArt pLatform Against paThogEns for CoronaVirus—Exscalate4CoV” founded by the EU’s H2020-SC1-PHE-CORONAVIRUS-2020 call, grant N. 101003551.