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posted on 2024-03-04, 22:24 authored by Prasad R. Kopparapu, Martin C. Pearce, Christiane V. Löhr, Cathy Duong, Hyo Sang Jang, Shanthakumar Tyavanagimatt, Edmond F. O'Donnell, Harikrishna Nakshatri, Siva K. Kolluri BFC1108 activates intrinsic mitochondrial death pathway. A, BFC1108 suppresses the colony-forming ability of MEF cells in a Bcl-2–dependent manner. WT and Bcl-2−/− MEF cells were treated for 48 hours in a medium containing 10% FBS and the colony formation was determined after 2 weeks. B, Quantification of colony formation data shown in A. **, P < 0.01. C, Bax or Bak is required for BFC1108-induced suppression of viability. WT MEF, Bax Knockout (Bax−/− Bak+/+), Bak knockout (Bax+/+ Bak−/−) and double knockout (Bax−/− Bak−/−) MEF cells were treated with 1 µmol/L BFC1108 for 24 hours in 10% FBS medium and viability was assessed using CellTiter-Glo assay. **, P < 0.01; ***, P < 0.001; ns, not significant. D, BFC1108 decreases mitochondrial membrane potential of Bcl-2–expressing H460 lung cancer cells. JC-1 dye was used to stain live H460 cells that were treated with 10 µmol/L BFC1108 for 16 hours in 10% FBS containing medium. Images taken with FITC, and rhodamine filters were overlaid. Cells stained orange have intact mitochondrial outer membrane and the ones turning green have compromised outer membrane indicating loss of membrane potential.
Funding
HHS | NIH | National Cancer Institute (NCI)
DOD | USA | MEDCOM | Congressionally Directed Medical Research Programs (CDMRP)
U.S. Department of Agriculture (USDA)
History
ARTICLE ABSTRACT
Cancer cells exploit the expression of anti-apoptotic protein Bcl-2 to evade apoptosis and develop resistance to therapeutics. High levels of Bcl-2 leads to sequestration of pro-apoptotic proteins causing the apoptotic machinery to halt. In this study, we report discovery of a small molecule, BFC1108 (5-chloro-N-(2-ethoxyphenyl)-2-[(4-methoxybenzyol)amino]benzamide), which targets Bcl-2 and converts it into a pro-apoptotic protein. The apoptotic effect of BFC1108 is not inhibited, but rather potentiated, by Bcl-2 overexpression. BFC1108 induces a conformational change in Bcl-2, resulting in the exposure of its BH3 domain both in vitro and in vivo. BFC1108 suppresses the growth of triple-negative breast cancer xenografts with high Bcl-2 expression and inhibits breast cancer lung metastasis. This study demonstrates a novel approach to targeting Bcl-2 using BFC1108, a small molecule Bcl-2 functional converter that effectively induces apoptosis in Bcl-2–expressing cancers.
We report the identification of a small molecule that exposes the Bcl-2 killer conformation and induces death in Bcl-2–expressing cancer cells. Selective targeting of Bcl-2 and elimination of cancer cells expressing Bcl-2 opens up new therapeutic avenues.
