RESUMO
Hsp90 encodes a ubiquitous molecular chaperone protein conserved among species which acts on multiple substrates, many of which are important cell-signaling proteins. Inhibition of Hsp90 function has been promoted as a mechanism to degrade client proteins involved in tumorigenesis and disease progression. Several assays to monitor inhibition of Hsp90 function currently exist but are limited in their use for a drug discovery campaign. Using data from the crystal structure of an initial hit compound, we have developed a fluorescence polarization assay to monitor binding of compounds to the ATP-binding site of Hsp90. This assay is very robust (Z' > 0.9) and can detect affinity of compounds with IC50s to 40 nM. We have used this assay in conjunction with cocrystal structures of small molecules to drive a structure-based design program aimed at the discovery and optimization of a novel class of potent Hsp90 inhibitors.
Assuntos
Cumarínicos/química , Cumarínicos/farmacologia , Polarização de Fluorescência/métodos , Proteínas de Choque Térmico HSP90/antagonistas & inibidores , Pirazóis/química , Pirazóis/farmacologia , Adenosina Trifosfatases/antagonistas & inibidores , Sítios de Ligação , Cristalografia por Raios X , Concentração Inibidora 50 , Resorcinóis/química , Saccharomyces cerevisiae/enzimologia , Relação Estrutura-AtividadeRESUMO
A range of N-substituted isoindolin-1-ones was prepared and their potential as novel antimicrobial agents was investigated. MIC values for active compounds were determined and reported.