RESUMO
The identification of a novel hit compound inhibitor of the protein-protein interaction between the influenza RNA-polymerase PA and PB1 subunits has been accomplished by means of high-throughput screening. A small family of structurally related molecules has been synthesized and biologically evaluated with most of the compounds showing micromolar potency of inhibition against viral replication.
Assuntos
Antivirais/toxicidade , Benzoxazóis/química , RNA Polimerases Dirigidas por DNA/metabolismo , Inibidores Enzimáticos/síntese química , Vírus da Influenza A/efeitos dos fármacos , Animais , Antivirais/síntese química , Antivirais/química , Benzoxazóis/síntese química , Benzoxazóis/toxicidade , RNA Polimerases Dirigidas por DNA/química , Cães , Avaliação Pré-Clínica de Medicamentos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/toxicidade , Vírus da Influenza A/enzimologia , Células Madin Darby de Rim Canino , Domínios e Motivos de Interação entre Proteínas/efeitos dos fármacos , Subunidades Proteicas/química , Subunidades Proteicas/metabolismo , Relação Estrutura-AtividadeRESUMO
A fast and efficient protocol for the generation of substituted 5-arylidene rhodanines in a sequential one-pot two-step process combining the "Holmberg method" and the Knoevenagel condensation under microwave-assisted conditions has been developed. The final compounds 11a-k have been obtained in high yield and purity after a simple precipitation from methanol, making this procedure facile, practical, and rapid to execute.
Assuntos
Técnicas de Química Combinatória/métodos , Micro-Ondas , Rodanina/química , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Fatores de TempoRESUMO
The influenza RNA polymerase complex, which consists of the three subunits PA, PB1, and PB2, is a promising target for the development of new antiviral drugs. A large library of benzofurazan compounds was synthesized and assayed against influenza virus A/WSN/33 (H1N1). Most of the new derivatives were found to act by inhibiting the viral RNA polymerase complex through disruption of the complex formed between subunits PA and PB1. Docking studies were also performed to elucidate the binding mode of benzofurazans within the PB1 binding site in PA and to identify amino acids involved in their mechanism of action. The predicted binding pose is fully consistent with the biological data and lays the foundation for the rational development of more effective PA-PB1 inhibitors.
Assuntos
Antivirais/síntese química , Benzoxazóis/química , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , Inibidores Enzimáticos/síntese química , Vírus da Influenza A Subtipo H1N1/enzimologia , Proteínas Virais/antagonistas & inibidores , Antivirais/farmacologia , Benzoxazóis/farmacologia , Sítios de Ligação , RNA Polimerases Dirigidas por DNA/metabolismo , Inibidores Enzimáticos/farmacologia , Células HEK293 , Humanos , Simulação de Dinâmica Molecular , Ligação Proteica , Mapas de Interação de Proteínas/efeitos dos fármacos , Estrutura Terciária de Proteína , Subunidades Proteicas/antagonistas & inibidores , Subunidades Proteicas/metabolismo , Proteínas Virais/metabolismo , Replicação Viral/efeitos dos fármacosRESUMO
A hit optimization protocol applied to the first nonnucleoside inhibitor of the ATPase activity of human DEAD-box RNA helicase DDX3 led to the design and synthesis of second-generation rhodanine derivatives with better inhibitory activity toward cellular DDX3 and HIV-1 replication. Additional DDX3 inhibitors were identified among triazine compounds. Biological data were rationalized in terms of structure-activity relationships and docking simulations. Antiviral activity and cytotoxicity of selected DDX3 inhibitors are reported and discussed. A thorough analysis confirmed human DDX3 as a valid anti-HIV target. The compounds described herein represent a significant advance in the pursuit of novel drugs that target HIV-1 host cofactors.