作者: Thompson, Connor A H ; Schulz, Annika Lea ; Lemieux, Gabriel ; Gao, Guang ; Gordon, Danielle G ; Désilets, Antoine ; Leduc, Richard ; Jean, François ; Nabi, Ivan Robert ; Pérez-Vargas, Jimena ; Boudreault, Pierre-Luc ; Krajden, Mel ; Ennis, Siobhan ; Niikura, Masahiro

SARS-CoV-2 Omicron subvariants with increased transmissibility and immune evasion are spreading globally with alarming persistence. Whether the mutations and evolution of spike (S) Omicron subvariants alter the viral hijacking of human TMPRSS2 for viral entry remains to be elucidated. This is particularly important to investigate because of the large number and diversity of mutations of S Omicron subvariants reported since the emergence of BA.1. Here we report that human TMPRSS2 is a molecular determinant of viral entry for all the Omicron clinical isolates tested in human lung cells, including ancestral Omicron subvariants (BA.1, BA.2, BA.5), contemporary Omicron subvariants (BQ.1.1, XBB.1.5, EG.5.1) and currently circulating Omicron BA.2.86. First, we used a co-transfection assay to demonstrate the endoproteolytic cleavage by TMPRSS2 of spike Omicron subvariants. Second, we found that N-0385, a highly potent TMPRSS2 inhibitor, is a robust entry inhibitor of virus-like particles harbouring the S protein of Omicron subvariants. Third, we show that N-0385 exhibits nanomolar broad-spectrum antiviral activity against live Omicron subvariants in human Calu-3 lung cells and primary patient-derived bronchial epithelial cells. Interestingly, we found that N-0385 is 10-20 times more potent than the repositioned TMPRSS2 inhibitor, camostat, against BA.5, EG.5.1, and BA.2.86. We further found that N-0385 shows broad synergistic activity with clinically approved direct-acting antivirals (DAAs), i.e., remdesivir and nirmatrelvir, against Omicron subvariants, demonstrating the potential therapeutic benefits of a multi-targeted treatment based on N-0385 and DAAs.

2023-12-08·Emerging Microbes & Infections

A novel class of broad-spectrum active-site-directed 3C-like protease inhibitors with nanomolar antiviral activity against highly immune-evasive SARS-CoV-2 Omicron subvariants

Article

作者: Smith, Jason R. ; Ton, Anh-Tien ; Worrall, Liam J. ; Boudreault, Pierre-Luc ; Pérez-Vargas, Jimena ; Hong, Christopher Yee ; Young, Robert N. ; Leduc, Richard ; Kovacic, Suzana ; Gordon, Danielle G. ; Thompson, Connor A. H. ; Ennis, Siobhan ; Ban, Fuqiang ; Shapira, Tirosh ; Villanueva, Ivan ; Kenward, Calem ; Bielecki, Michael ; Jean, François ; Olmstead, Andrea D. ; Dudek, Svenja ; Krajden, Mel ; Levett, Paul N. ; Paetzel, Mark ; Vuckovic, Marija ; De Guzman, Joshua ; Cherkasov, Artem ; Lee, Jaeyong ; Niikura, Masahiro ; Gang, Shutong ; Strynadka, Natalie C. J.

Antivirals with broad coronavirus activity are important for treating high-risk individuals exposed to the constantly evolving SARS-CoV-2 variants of concern (VOCs) as well as emerging drug-resistant variants. We developed and characterized a novel class of active-site-directed 3-chymotrypsin-like protease (3CLpro) inhibitors (C2-C5a). Our lead direct-acting antiviral (DAA), C5a, is a non-covalent, non-peptide with a dissociation constant of 170 nM against recombinant SARS-CoV-2 3CLpro. The compounds C2-C5a exhibit broad-spectrum activity against Omicron subvariants (BA.5, BQ.1.1, and XBB.1.5) and seasonal human coronavirus-229E infection in human cells. Notably, C5a has median effective concentrations of 30-50 nM against BQ.1.1 and XBB.1.5 in two different human cell lines. X-ray crystallography has confirmed the unique binding modes of C2-C5a to the 3CLpro, which can limit virus cross-resistance to emerging Paxlovid-resistant variants. We tested the effect of C5a with two of our newly discovered host-directed antivirals (HDAs): N-0385, a TMPRSS2 inhibitor, and bafilomycin D (BafD), a human vacuolar H⁺-ATPase [V-ATPase] inhibitor. We demonstrated a synergistic action of C5a in combination with N-0385 and BafD against Omicron BA.5 infection in human Calu-3 lung cells. Our findings underscore that a SARS-CoV-2 multi-targeted treatment for circulating Omicron subvariants based on DAAs (C5a) and HDAs (N-0385 or BafD) can lead to therapeutic benefits by enhancing treatment efficacy. Furthermore, the high-resolution structures of SARS-CoV-2 3CLpro in complex with C2-C5a will facilitate future rational optimization of our novel broad-spectrum active-site-directed 3C-like protease inhibitors.

2023-01-01·Antiviral Research

Discovery of lead natural products for developing pan-SARS-CoV-2 therapeutics

Article

作者: Jaidee, Wuttichai ; Boudreault, Pierre-Luc ; Laphookhieo, Surat ; Kanokmedhakul, Kwanjai ; Olmstead, Andrea D ; Niikura, Masahiro ; Freire, Vitor F ; Leduc, Richard ; Suthiphasilp, Virayu ; Williams, David E ; Berlinck, Roberto G S ; Chin, Aaleigha ; Pérez-Vargas, Jimena ; Bernardi, Darlon I ; Yenjai, Chavi ; Villanueva, Ivan ; Jean, François ; De Guzman, Joshua ; Nuzzo, Genoveffa ; Fontana, Angelo ; Wang, Meng ; Chaiyosang, Boonyanoot ; Levett, Paul ; Tietjen, Ian ; Nabi, Ivan Robert ; Polbuppha, Isaraporn ; Manzo, Emiliano ; Teles, Helder Lopes ; Gubiani, Juliana R ; Cheenpracha, Sarot ; Shi, Pei-Yong ; Xie, Xuping ; Cherkasov, Artem ; Meesakul, Pornphimol ; Raksat, Achara ; Shapira, Tirosh ; Gao, Guang ; Thompson, Connor A H ; Bautista-Sánchez, Diana ; Ennis, Siobhan ; Quintana-Bulla, Jairo I ; Kanokmedhakul, Somdej ; Andersen, Raymond J ; Krajden, Mel ; Agafitei, Olga

The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a global public health crisis. The reduced efficacy of therapeutic monoclonal antibodies against emerging SARS-CoV-2 variants of concern (VOCs), such as omicron BA.5 subvariants, has underlined the need to explore a novel spectrum of antivirals that are effective against existing and evolving SARS-CoV-2 VOCs. To address the need for novel therapeutic options, we applied cell-based high-content screening to a library of natural products (NPs) obtained from plants, fungi, bacteria, and marine sponges, which represent a considerable diversity of chemical scaffolds. The antiviral effect of 373 NPs was evaluated using the mNeonGreen (mNG) reporter SARS-CoV-2 virus in a lung epithelial cell line (Calu-3). The screening identified 26 NPs with half-maximal effective concentrations (EC₅₀) below 50 μM against mNG-SARS-CoV-2; 16 of these had EC₅₀ values below 10 μM and three NPs (holyrine A, alotaketal C, and bafilomycin D) had EC₅₀ values in the nanomolar range. We demonstrated the pan-SARS-CoV-2 activity of these three lead antivirals against SARS-CoV-2 highly transmissible Omicron subvariants (BA.5, BA.2 and BA.1) and highly pathogenic Delta VOCs in human Calu-3 lung cells. Notably, holyrine A, alotaketal C, and bafilomycin D, are potent nanomolar inhibitors of SARS-CoV-2 Omicron subvariants BA.5 and BA.2. The pan-SARS-CoV-2 activity of alotaketal C [protein kinase C (PKC) activator] and bafilomycin D (V-ATPase inhibitor) suggest that these two NPs are acting as host-directed antivirals (HDAs). Future research should explore whether PKC regulation impacts human susceptibility to and the severity of SARS-CoV-2 infection, and it should confirm the important role of human V-ATPase in the VOC lifecycle. Interestingly, we observed a synergistic action of bafilomycin D and N-0385 (a highly potent inhibitor of human TMPRSS2 protease) against Omicron subvariant BA.2 in human Calu-3 lung cells, which suggests that these two highly potent HDAs are targeting two different mechanisms of SARS-CoV-2 entry. Overall, our study provides insight into the potential of NPs with highly diverse chemical structures as valuable inspirational starting points for developing pan-SARS-CoV-2 therapeutics and for unravelling potential host factors and pathways regulating SARS-CoV-2 VOC infection including emerging omicron BA.5 subvariants.

100 项与 N-0385 相关的药物交易

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