RESUMEN
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.
Asunto(s)
Cumarinas/química , Cumarinas/farmacología , Polarización de Fluorescencia/métodos , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Pirazoles/química , Pirazoles/farmacología , Adenosina Trifosfatasas/antagonistas & inhibidores , Sitios de Unión , Cristalografía por Rayos X , Concentración 50 Inhibidora , Resorcinoles/química , Saccharomyces cerevisiae/enzimología , Relación Estructura-ActividadRESUMEN
The in vitro properties of two peptide aldehydes are described which relate to their potential use as elastase inhibitors in the treatment of emphysema. When compared with alpha 1-antitrypsin they demonstrate potential advantages in terms of potency, specificity and stability to cigarette smoke.
