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  1. Article ; Online: Amino Acid Polymorphisms on the Brazilian Strain of Yellow Fever Virus Methyltransferase Are Related to the Host's Immune Evasion Mediated by Type I Interferon.

    Furtado, Nathália Dias / de Mello, Iasmim Silva / de Godoy, Andre Schutzer / Noske, Gabriela Dias / Oliva, Glaucius / Canard, Bruno / Decroly, Etienne / Bonaldo, Myrna C

    Viruses

    2023  Volume 15, Issue 1

    Abstract: Since late 2016, a yellow fever virus (YFV) variant carrying a set of nine amino acid variations has circulated in South America. Three of them were mapped on the methyltransferase (MTase) domain of viral NS5 protein. To assess whether these changes ... ...

    Abstract Since late 2016, a yellow fever virus (YFV) variant carrying a set of nine amino acid variations has circulated in South America. Three of them were mapped on the methyltransferase (MTase) domain of viral NS5 protein. To assess whether these changes affected viral infectivity, we synthesized YFV carrying the MTase of circulating lineage as well as its isoform with the residues of the previous strains (NS5 K101R, NS5 V138I, and NS5 G173S). We observed a slight difference in viral growth properties and plaque phenotype between the two synthetic YFVs. However, the MTase polymorphisms associated with the Brazilian strain of YFV (2016-2019) confer more susceptibility to the IFN-I. In addition, in vitro MTase assay revealed that the interaction between the YFV MTase and the methyl donor molecule (SAM) is altered in the Brazilian MTase variant. Altogether, the results reported here describe that the MTase carrying the molecular signature of the Brazilian YFV circulating since 2016 might display a slight decrease in its catalytic activity but virtually no effect on viral fitness in the parameters comprised in this study. The most marked influence of these residues stands in the immune escape against the antiviral response mediated by IFN-I.
    MeSH term(s) Yellow fever virus/physiology ; Interferon Type I/genetics ; Amino Acids ; Immune Evasion ; Brazil ; Methyltransferases/metabolism ; Viral Nonstructural Proteins/genetics
    Chemical Substances Interferon Type I ; Amino Acids ; Methyltransferases (EC 2.1.1.-) ; Viral Nonstructural Proteins
    Language English
    Publishing date 2023-01-10
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2516098-9
    ISSN 1999-4915 ; 1999-4915
    ISSN (online) 1999-4915
    ISSN 1999-4915
    DOI 10.3390/v15010191
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways.

    Ruiz, Uriel Enrique Aquino / Santos, Igor Andrade / Grosche, Victória Riquena / Fernandes, Rafaela Sachetto / de Godoy, Andre Schutzer / Torres, Jhoan David Aguillón / Freire, Marjorie Caroline Liberato Cavalcanti / Mesquita, Nathalya Cristina de Moraes Roso / Guevara-Vega, Marco / Nicolau-Junior, Nilson / Sabino-Silva, Robinson / Mineo, Tiago Wilson Patriarca / Oliva, Glaucius / Jardim, Ana Carolina Gomes

    Virus research

    2022  Volume 324, Page(s) 199029

    Abstract: The Chikungunya virus (CHIKV) causes Chikungunya fever, a disease characterized by symptoms such as arthralgia/polyarthralgia. Currently, there are no antivirals approved against CHIKV, emphasizing the need to develop novel therapies. The ... ...

    Abstract The Chikungunya virus (CHIKV) causes Chikungunya fever, a disease characterized by symptoms such as arthralgia/polyarthralgia. Currently, there are no antivirals approved against CHIKV, emphasizing the need to develop novel therapies. The imidazonaphthyridine compound (RO8191), an interferon-α (IFN-α) agonist, was reported as a potent inhibitor of HCV. Here RO8191 was investigated for its potential to inhibit CHIKV replication in vitro. RO8191 inhibited CHIKV infection in BHK-21 and Vero-E6 cells with a selectivity index (SI) of 12.3 and 37.3, respectively. Additionally, RO8191 was capable to protect cells against CHIKV infection, inhibit entry by virucidal activity, and strongly impair post-entry steps of viral replication. An effect of RO8191 on CHIKV replication was demonstrated in BHK-21 through type-1 IFN production mechanism and in Vero-E6 cells which has a defective type-1 IFN production, also suggesting a type-1 IFN independent mode of action. Molecular docking calculations demonstrated interactions of RO8191 with the CHIKV E proteins, corroborated by the ATR-FTIR assay, and with non-structural proteins, supported by the CHIKV-subgenomic replicon cells assay.
    MeSH term(s) Animals ; Chlorocebus aethiops ; Humans ; Chikungunya virus ; Chikungunya Fever/drug therapy ; Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Molecular Docking Simulation ; Virus Replication ; Vero Cells ; Interferon Type I/pharmacology
    Chemical Substances Antiviral Agents ; Interferon Type I
    Language English
    Publishing date 2022-12-22
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 605780-9
    ISSN 1872-7492 ; 0168-1702
    ISSN (online) 1872-7492
    ISSN 0168-1702
    DOI 10.1016/j.virusres.2022.199029
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Discovery of an imidazonaphthyridine and a riminophenazine as potent anti-Zika virus agents through a replicon-based high-throughput screening.

    Fernandes, Rafaela Sachetto / de Godoy, Andre Schutzer / Santos, Igor Andrade / Noske, Gabriela Dias / de Oliveira, Ketllyn Irene Zagato / Gawriljuk, Victor Oliveira / Gomes Jardim, Ana Carolina / Oliva, Glaucius

    Virus research

    2021  Volume 299, Page(s) 198388

    Abstract: The 2015/16 Zika virus (ZIKV) epidemic led to almost 1 million confirmed cases in 84 countries and was associated to the development of congenital microcephaly and Guillain-Barré syndrome. More recently, a ZIKV African lineage was identified in Brazil ... ...

    Abstract The 2015/16 Zika virus (ZIKV) epidemic led to almost 1 million confirmed cases in 84 countries and was associated to the development of congenital microcephaly and Guillain-Barré syndrome. More recently, a ZIKV African lineage was identified in Brazil raising concerns about a future outbreak. The long-term consequences of viral infection emphasizes the need for the development of effective anti-ZIKV drugs. In this study, we developed and characterized a ZIKV replicon cell line for the screening of viral replication inhibitors. The replicon system was developed by engineering the IRES-Neo cassette into the 3' UTR terminus of the ZIKV Rluc DNA construct. After in vitro transcription, replicon RNA was used to transfect BHK-21 cells, that were selected with G418, thus generating the BHK-21-RepZIKV_IRES-Neo cell line. Through this replicon-based cell system, we identified two molecules with potent anti-ZIKV activities, an imidazonaphthyridine and a riminophenazine, both from the MMV/DNDi Pandemic Response Box library of 400 drug-like compounds. The imidazonaphthyridine, known as RO8191, showed remarkable selectivity against ZIKV, while the riminophenazine, the antibiotic Clofazimine, could act as a non-nucleoside analog inhibitor of viral RNA-dependent RNA polymerase (RdRp), as evidenced both in vitro and in silico. The data showed herein supports the use of replicon-based assays in high-throughput screening format as a biosafe and reliable tool for antiviral drug discovery.
    MeSH term(s) Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Clofazimine/pharmacology ; Clofazimine/therapeutic use ; High-Throughput Screening Assays ; Humans ; Replicon ; Virus Replication ; Zika Virus/physiology ; Zika Virus Infection
    Chemical Substances Antiviral Agents ; Clofazimine (D959AE5USF)
    Language English
    Publishing date 2021-04-20
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 605780-9
    ISSN 1872-7492 ; 0168-1702
    ISSN (online) 1872-7492
    ISSN 0168-1702
    DOI 10.1016/j.virusres.2021.198388
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Structural and mechanistic insight from antiviral and antiparasitic enzyme drug targets for tropical infectious diseases.

    de Godoy, Andre Schutzer / Sachetto Fernandes, Rafaela / Campos Aguiar, Anna Caroline / Vieira Bueno, Renata / de Moraes Roso Mesquita, Nathalya Cristina / Carvalho Guido, Rafael Victorio / Oliva, Glaucius

    Current opinion in structural biology

    2019  Volume 59, Page(s) 65–72

    Abstract: With almost half of the world population living at risk, tropical infectious diseases cause millions of deaths every year in developing countries. Considering the lack of economic prospects for investment in this field, approaches aiming the rational ... ...

    Abstract With almost half of the world population living at risk, tropical infectious diseases cause millions of deaths every year in developing countries. Considering the lack of economic prospects for investment in this field, approaches aiming the rational design of compounds, such as structure-based drug discovery (SBDD), fragment screening, target-based drug discovery, and drug repurposing are of special interest. Herein, we focused in the advances on the field of SBDD targeting arboviruses such as dengue, yellow fever, zika and chikungunya enzymes of the RNA replication complex (RC) and enzymes involved in a variety of pathways essential to ensure parasitic survival in the host, for malaria, Chagas e leishmaniasis diseases. We also highlighted successful examples such as promising new inhibitors and molecules already in preclinical/clinical phase tests, major gaps in the field and perspectives for the future of drug design for tropical diseases.
    MeSH term(s) Antiparasitic Agents/chemistry ; Antiparasitic Agents/pharmacology ; Antiviral Agents/chemistry ; Antiviral Agents/pharmacology ; Enzyme Inhibitors/chemistry ; Enzyme Inhibitors/pharmacology ; Enzymes/chemistry ; Folic Acid Antagonists/chemistry ; Folic Acid Antagonists/pharmacology ; Humans ; Models, Molecular ; Molecular Conformation ; Protein Binding ; Protozoan Proteins/chemistry ; Protozoan Proteins/metabolism ; Quantitative Structure-Activity Relationship ; Tetrahydrofolate Dehydrogenase/chemistry ; Viral Proteins/chemistry ; Viral Proteins/metabolism
    Chemical Substances Antiparasitic Agents ; Antiviral Agents ; Enzyme Inhibitors ; Enzymes ; Folic Acid Antagonists ; Protozoan Proteins ; Viral Proteins ; Tetrahydrofolate Dehydrogenase (EC 1.5.1.3)
    Language English
    Publishing date 2019-04-05
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1068353-7
    ISSN 1879-033X ; 0959-440X
    ISSN (online) 1879-033X
    ISSN 0959-440X
    DOI 10.1016/j.sbi.2019.02.014
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: A Crystallographic Snapshot of SARS-CoV-2 Main Protease Maturation Process

    Noske, Gabriela / Nakamura, Aline / Gawriljuk, Victor / Fernandes, Rafaela / Lima, Gustavo / Rosa, Higor / Pereira, Humberto / Zeri, Ana / Nascimento, Andrey / Freire, Marjorie / Oliva, Glaucius / de Godoy, Andre Schutzer

    bioRxiv

    Abstract: SARS-CoV-2 is the causative agent of COVID-19. The dimeric form of the viral main protease is responsible for the cleavage of the viral polyprotein in 11 sites, including its own N and C-terminus. Herein, we used X-ray crystallography to characterize an ... ...

    Abstract SARS-CoV-2 is the causative agent of COVID-19. The dimeric form of the viral main protease is responsible for the cleavage of the viral polyprotein in 11 sites, including its own N and C-terminus. Herein, we used X-ray crystallography to characterize an immature form of the main protease. We propose that this form preludes the cis-cleavage of N-terminal residues within the dimer, leading to the mature active site. Using fragment screening, we probe new cavities in this form which can be used to guide therapeutic development. Furthermore, we characterized a serine site-directed mutant of the main protease bound to its endogenous N and C-terminal residues during the formation of the tetramer. This quaternary form is also present in solution, suggesting a transitional state during the C-terminal trans-cleavage.
    Keywords covid19
    Language English
    Publishing date 2020-12-23
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2020.12.23.424149
    Database COVID19

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  6. Article ; Online: Pyronaridine Protects Against SARS-CoV-2 in Mouse

    Puhl, Ana C. / Gomes, Giovanni F. / Damasceno, Samara / de Godoy, Andre Schutzer / Noske, Gabriela D. / Nakamura, Aline M. / Gawrijuk, Victor O. / Fernandes, Rafaela S. / Monakhova, Natalia / Riabova, Olga / Lane, Thomas R / Makarov, Vadim / Veras, Flavio P. / Batah, Sabrina S. / Fabro, Alexandre T. / Oliva, Glaucius / Cunha, Fernando / Alves-Filho, Jose C. / Cunha, Thiago M. /
    Ekins, Sean

    bioRxiv

    Abstract: There are currently relatively few small-molecule antiviral drugs that are either approved or emergency approved for use against SARS-CoV-2. One of these is remdesivir, which was originally repurposed from its use against Ebola and functions by causing ... ...

    Abstract There are currently relatively few small-molecule antiviral drugs that are either approved or emergency approved for use against SARS-CoV-2. One of these is remdesivir, which was originally repurposed from its use against Ebola and functions by causing early RNA chain termination. We used this as justification to evaluate three molecules we had previously identified computationally with antiviral activity against Ebola and Marburg. Out of these we previously identified pyronaridine, which inhibited the SARS-CoV-2 replication in A549-ACE2 cells. Herein, the in vivo efficacy of pyronaridine has now been assessed in a K18-hACE transgenic mouse model of COVID-19. Pyronaridine treatment demonstrated a statistically significant reduction of viral load in the lungs of SARS CoV-2 infected mice. Furthermore, the pyronaridine treated group reduced lung pathology, which was also associated with significant reduction in the levels of pro-inflammatory cytokines/chemokine and cell infiltration. Notably, pyronaridine inhibited the viral PLpro activity in vitro (IC50 of 1.8 uM) without any effect on Mpro, indicating a possible molecular mechanism involved in its ability to inhibit SARS-CoV-2 replication. Interestingly, pyronaridine also selectively inhibits the host kinase CAMK1 (IC50 of 2.4 uM). We have also generated several pyronaridine analogs to assist in understanding the structure activity relationship for PLpro inhibition. Our results indicate that pyronaridine is a potential therapeutic candidate for COVID-19.
    Keywords covid19
    Language English
    Publishing date 2021-09-30
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2021.09.30.462449
    Database COVID19

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