Design and Testing of Bi-functional, P-loop Targeted MDM2 Inhibitors
Abstract
Our proposal is to design and evaluate a novel class of bifunctional MDM2 inhibitors, based on the discovery that nucleotides can bind to the P-loop of MDM2 and cause its relocalization to the nucleolus. Such bifunctional compounds will be designed to target MDM2, but not other P-loop-containing proteins. This approach represents a new strategy for the inhibition of MDM2 function and the treatment of breast cancer. During the second year of this grant we have used the cloned, expressed, and purified GST-fused Mdm2 wild-type and C-terminally deleted RING domain protein to continue to test all commercially and privately available ATP analogs (including fluorescent analogs) for binding to Mdm2 and have further insight into the features of ATP required for binding to Mdm2. We have also (1) determined that crosslinking to 8-azido ATP inhibits ATP binding and E3 ligase activity; (2) developed high-throughput autoubiquitination assay to discover Mdm2 ligase inhibitors (3) developed a high-throughput docking assay based on Mdm2 s RING domain structure and (4) developed a high-throughput compatible luciferase-based competition assay for compounds that bind to Mdm2.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 2008
- Accession Number
- ADA573755
Entities
People
- Brent R Stockwell
- Carol L. Prives
Organizations
- Columbia University