Imaging of protease activity in live cells using activity based probes

US8968700B2 · kind B2 · utility

Assignee

• The Board of Trustees of the Leland Stanford Junior University · US

Inventors

• Matthew S. Bogyo · Redwood City, US
• Galia Blum · Jerusalem, IL
• Georges Von Degenfeld · Leverkusen, DE

Key dates

• Filing date: Aug 10, 2006
• Grant date: Mar 3, 2015
• Expiry date: Jul 20, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K5/06078
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Methods and materials for the imaging of cells containing active proteases such as cathepsin are disclosed. The present materials include activity based probes that bind to an enzyme and are subsequently cleaved. Cleavage results in a fluorescent signal due to removal of a quenching group which, when present on the probe causes altered or no fluorescence. The probes employ an acyloxymethyl ketone reactive group, one or more amino acids for determining specificity, a fluorophore and a quencher. The probes are cell permeable and may use, for example, a QSY7 (diarylyrhodamine) quencher and a BODIPY (bora-diaza-indecene) dye.