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Sci Rep ; 6: 33477, 2016 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-27633629

RESUMO

HIV-1 integrase (IN) plays an essential role in viral replication and thus serves as an important target for chemotherapeutic intervention against HIV-1 infection. However, the current three clinical IN inhibitors, raltegravir, elvitegravir and dolutegravir share the same inhibitory mechanism, resulting in a common clinical resistance profile which have emerged in infected patients receiving treatment. Therefore, it is important to develop small molecule inhibitors that impair IN function with distinct mechanisms of action. In this work, a magnetic-beads based biochemical assay targeting the protein-protein interaction (PPI) between HIV IN and the cellular cofactor LEDGF/p75 was developed for identification of HIV-1 IN inhibitors. Furthermore, a library containing 1000 US. Food and Drug Administration (FDA)-approved drugs currently used for human medication was screened to identify inhibitors targeting the PPI. The assay was proved to be quite robust and with the novel assay we successfully identified dexlansoprazole (IC50 of 4.8 µM), a FDA-approved proton pump inhibitor, as a potential inhibitor for the PPI between IN and LEDGF/p75, which bound to the LEDGF/p75 partner with a kinetic dissociation (Kd) constant of 330 nM ± 2.6 nM.


Assuntos
Bioensaio/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Integrase de HIV/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Magnetismo , Microesferas , Níquel/química , Sefarose/química , Dexlansoprazol/farmacologia , Ensaios de Triagem em Larga Escala , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/química , Interleucina-23/farmacologia , Peptídeos/farmacologia , Ligação Proteica/efeitos dos fármacos , Domínios Proteicos , Mapeamento de Interação de Proteínas , Inibidores da Bomba de Prótons/química , Inibidores da Bomba de Prótons/farmacologia , Reprodutibilidade dos Testes
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