Open AccessIdentification of Inhibitors for MDM2 Ubiquitin Ligase Activity from Natural Product Extracts by a Novel High-Throughput Electrochemiluminescent Screen
Author(s) -
Christy A. Sasiela,
David Stewart,
Jirouta Kitagaki,
Yassamin J. Safiran,
Yili Yang,
Allan M. Weissman,
Pankaj Oberoi,
Ilia V. Davydov,
Ekaterina I. Goncharova,
John A. Beutler,
James B. McMahon,
Barry R. O’Keefe
Publication year - 2008
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057108315038
Subject(s) - natural product , ubiquitin ligase , high throughput screening , drug discovery , ubiquitin , mdm2 , chemistry , computational biology , anticancer drug , bioassay , pharmacology , drug , biology , biochemistry , apoptosis , genetics , gene
High-throughput screening technologies have revolutionized the manner in which potential therapeutics are identified. Although they are the source of lead compounds for ~65% of anticancer and antimicrobial drugs approved by the Food and Drug Administration between 1981 and 2002, natural products have largely been excluded from modern screening programs. This is due, at least in part, to the inherent difficulties in testing complex extract mixtures, which often contain nuisance compounds, in modern bioassay systems. In this article, the authors present a novel electrochemiluminescent assay system for inhibition of MDM2 activity that is suitable for testing natural product extracts in high-throughput screening systems. The assay was used to screen more than 144,000 natural product extracts. The authors identified 1 natural product, sempervirine, that inhibited MDM2 auto-ubiquitination, MDM2-mediated p53 degradation, and led to accumulation of p53 in cells. Sempervirine preferentially induced apoptosis in transformed cells expressing wild-type p53, suggesting that it could be a potential lead for anticancer therapeutics.