Inhibition of SARS-CoV-2 3CLpro by chemically modified tyrosinase from <i>Agaricus bisporus</i>

Aguilera-Rodriguez, David ; Ortega-Alarcon, David ; Vazquez-Calvo, Angela ; Ricci, Veronica ; Abian, Olga (Universidad de Zaragoza) ; Velazquez-Campoy, Adrian (Universidad de Zaragoza) ; Alcami, Antonio ; Palomo, Jose M.

Resumen: Antiviral compounds are crucial to controlling the SARS-CoV-2 pandemic. Approved drugs have been tested for their efficacy against COVID-19, and new pharmaceuticals are being developed as a complementary tool to vaccines. In this work, a cheap and fast purification method for natural tyrosinase from Agaricus bisporus (AbTyr) fresh mushrooms was developed to evaluate the potential of this enzyme as a therapeutic protein via the inhibition of SARS-CoV-2 3CLpro protease activity in vitro. AbTyr showed a mild inhibition of 3CLpro. Thus, different variants of this protein were synthesized through chemical modifications, covalently binding different tailor-made glycans and peptides to the amino terminal groups of the protein. These new tyrosinase conjugates were purified and characterized through circular dichroism and fluorescence spectroscopy analyses, and their stability was evaluated under different conditions. Subsequently, all these tyrosinase conjugates were tested for 3CLpro protease inhibition. From them, the conjugate between tyrosinase and a dextran-aspartic acid (6 kDa) polymer showed the highest inhibition, with an IC50 of 2.5 μg ml−1 and IC90 of 5 μg ml−1, with no cytotoxicity activity by polymer insertion. Finally, SARS-CoV-2 virus infection was studied. It was found that this new AbTyr-Dext6000 protein showed an 80% decrease in viral load. These results show the capacity of these tyrosinase bioconjugates as potential therapeutic proteins, opening the possibility of extension and applicability against other different viruses.

Idioma: Inglés

DOI: 10.1039/d4md00289j

Año: 2024

Publicado en: RSC medicinal chemistry 15, 12 (2024), 4159-4167
ISSN: 2632-8682

Factor impacto JCR: 3.6 (2024)

Categ. JCR: CHEMISTRY, MEDICINAL rank: 32 / 72 = 0.444 (2024) - Q2 - T2
Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 129 / 319 = 0.404 (2024) - Q2 - T2

Factor impacto SCIMAGO: 0.841 - Pharmaceutical Science (Q1) - Organic Chemistry (Q1) - Pharmacology (Q2) - Drug Discovery (Q2) - Biochemistry (Q2) - Molecular Medicine (Q2)

Financiación: info:eu-repo/grantAgreement/ES/CSIC/PIE-202480E088
Financiación: info:eu-repo/grantAgreement/ES/CSIC/PTI-Global Health SGL2103036

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)