Category: Assay Development and Screening

1159-E - HTRF solution of identifying selective inhibitors of BCL-2 family

Wednesday, February 7, 2018
11:30 AM - 12:30 PM

Escape from apoptosis is a key attribute of tumor cells and facilitates chemo-resistance. Dynamic interaction between pro-apoptotic and pro-survival proteins in the BCL-2 family regulates apoptosis. The recent success of BCL-2 inhibitor, ABT-199 (Venetoclax) in chronic lymphocytic leukemia treatment is generating considerable interest in identifying selective inhibitors of BCL-2 family for novel anti-cancer therapies. HTRF (Homogeneous Time-resolved Fluorescence) is a FRET-based assay performed by a simple mix-and-read protocol that offers a wide range of applications in drug discovery research. Here, we reported two HTRF assays, BCL-2/BAK and BCL-XL/BAK protein-protein interactions, to identify selective inhibitors of BCL-2 family proteins. The assays were validated using four reference compounds, i.e. ABT-199, WEHI-539, ABT-263, and ABT-737. ABT-199, a selective inhibitor of BCL-2, exhibited ~150-fold higher activity towards BCL-2 (IC50=0.67 nM) than BCL-XL (IC50=98.7 nM). Meanwhile, WEHI-539, a selective inhibitor of BCL-XL, showed an IC50 of 1.76 nM to BCL-XL but IC50 > 200 mM to BCL-2. ABT-263 and ABT-737 are general inhibitors for both BCL-2 and BCL-XL and their activities were also confirmed in the current assays. Taken together, we successfully developed two HTRF assays to distinguish the compound specificity on BCL2 and BCL-XL, providing a simple and rapid method of identifying selective inhibitors of BCL-2 family.

Bing Xie

Managing Director of Asia Pacific
Cisbio Bioassays
Shanghai, Shanghai, China (People's Republic)

Bing Xie received his Ph.D. in Molecular Medicine from Yale University, School of Medicine. He then served as a postdoctoral associate in Department of Pathology, Yale University. In 2002, he became Director of Biological Application at Agilix Corporation. In 2006, Dr. Xie joined PerkinElmer as Technical Leader, focusing on the development of multiplexing platform in quantitative proteomics as well as the technologies in cell signaling and HTS. Subsequently, he worked at Helix Therapeutics as Director of R&D. Since 2008, Dr. Xie was leading Scientific Consulting Team in Cisbio, US. In the beginning of 2010, he became Managing Director, Asia Pacific for Cisbio Bioassays. Starting from 2013-2015, Dr. Xie was the member of Asia Council of The Society for Laboratory Automation and Screening (SLAS). Now he serves as Vice Chair for Executive Committee of Sino Drug Discovery Association.