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Doctoral Thesis
DOI
https://doi.org/10.11606/T.17.2020.tde-13042020-135944
Document
Author
Full name
Aline Lavado Tolardo
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2019
Supervisor
Committee
Figueiredo, Luiz Tadeu Moraes (President)
Acrani, Gustavo Olszanski
Santana, Rodrigo de Carvalho
Sousa, Ricardo Luiz Moro de
Title in Portuguese
Pesquisa de compostos com potencial ação antiviral para os vírus Mayaro e Oropouche
Keywords in Portuguese
Antivirais
Arbovírus
Mayaro
Oropouche
Abstract in Portuguese
Os Alphavirus e Orthobunyavirus são arbovírus importantes que causam grande impacto econômico e social no Brasil. Não existem antivirais disponíveis para o tratamento desses vírus. Portanto, é fundamental ter medicamentos terapêuticos que combatam os sintomas e sinais manifestados pela doença em casos leves e moderados. Neste trabalho, desenvolvemos um ensaio de luminescência baseada em células para a triagem de potenciais pequenas moléculas para os vírus Mayaro e Oropouche que mede o efeito citopático (CPE) induzido pelo vírus em células Vero usando o sistema CellTiter Glo baseado em luminescência. O ensaio foi validado no formato de placa de 384 poços e mostrou valores de Z maiores que 0,7, background maior que 30 e sinal-ruído maior que 10, demonstrando alta capacidade. Quatro bibliotecas de compostos foram testadas na concentração de 10 uM para ambos os testes. Identificamos cinco compostos que inibiram o efeito citopático induzido pelos vírus em > 50%, com valores de EC50/CC50 comparáveis aos determinados por outros ensaios baseados em células, validando assim a precisão e a capacidade do ensaio de servir como uma ferramenta para a descoberta de novos antivirais para os vírus Mayaro e Oropouche.
Title in English
Research on compounds with potential antiviral action for Mayaro and Oropouche viruses
Keywords in English
Antivirals
Arbovirus
Mayaro
Oropouche
Abstract in English
Alphavirus and Orthobunyavirus are important arboviruses that have a major economic and social impact in Brazil. There are no antivirals available to treat these viruses. Therefore, it is essential to have therapeutic drugs that combat the symptoms and signs manifested by the disease in mild and moderate cases. In this thesis, we developed a cell-based luminescence assay for the screening of potential small molecules for Mayaro and Oropouche viruses that measures the virus-induced cytopathic effect (CPE) in Vero cells (ATCC CCL81) using the luminescence-based CellTiter Glo system. The assay was validated in a 384-well plate format and showed Z values greater than 0.7, background greater than 30 and signal-to-noise greater than 10, demonstrating high capacity. Four compound libraries were tested at a concentration of 10 µM for both tests. We identified five compounds that inhibited virus-induced cytopathic effect by> 50%, with EC50/CC50 values comparable to those determined by other cell-based assays, thus validating the assay's accuracy and ability to serve as a tool for discovery of new antivirals for Mayaro and Oropouche viruses.
 
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Publishing Date
2020-05-05
 
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