Chemically Linked Vemurafenib Inhibitors Promote an Inactive BRAFV600E Conformation
Abstract
The BRAF kinase, within the mitogen activated protein kinase (MAPK) signaling pathway, harbors activating mutations in about half of melanomas and to a significant extent in many other cancers. A single valine to glutamic acid substitution at residue 600 (BRAFV600E) accounts for about 90% of these activating mutations. While BRAFV600E-selective small molecule inhibitors, such as debrafenib and vemurafenib, have shown therapeutic benefit, almost all patients develop resistance. Resistance often arises through reactivation of the MAPK pathway, typically through mutation of upstream RAS, downstream MEK, or splicing variants. RAF kinases signal as homo- and heterodimers, and another complication associated with small molecule BRAFV600E inhibition is drug-induced allosteric activation of a wild-type RAF subunit (BRAF or CRAF) of the kinase dimer, a process called “transactivation” or “paradoxical activation.” Here, we used BRAFV600E and vemurafenib as a model system to develop chemically linked kinase inhibitors to lock RAF dimers in an inactive conformation that cannot undergo transactivation. This structure-based design effort resulted in the development of Vem-BisAmide-2, a compound containing two vemurafenib molecules connected by a bis amide linker. We show that Vem-BisAmide-2 has comparable inhibitory potency as vemurafenib to BRAFV600E both in vitro and in cells but promotes an inactive dimeric BRAFV600Emore »
- Authors:
-
- Univ. of Pennsylvania, Philadelphia, PA (United States)
- Wistar Inst., Philadelphia, PA (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- National Institutes of Health (NIH); USDOE Office of Science (SC)
- OSTI Identifier:
- 1331701
- Grant/Contract Number:
- CA114046; K01CA175269; CA016520; GM103403)
- Resource Type:
- Accepted Manuscript
- Journal Name:
- ACS Chemical Biology
- Additional Journal Information:
- Journal Volume: 11; Journal Issue: 10; Journal ID: ISSN 1554-8929
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Peptides and proteins; Crystal structure; Oligomers; Inhibitors; Conformation
Citation Formats
Grasso, Michael, Estrada, Michelle A., Ventocilla, Christian, Samanta, Minu, Maksimoska, Jasna, Villanueva, Jessie, Winkler, Jeffrey D., and Marmorstein, Ronen. Chemically Linked Vemurafenib Inhibitors Promote an Inactive BRAFV600E Conformation. United States: N. p., 2016.
Web. doi:10.1021/acschembio.6b00529.
Grasso, Michael, Estrada, Michelle A., Ventocilla, Christian, Samanta, Minu, Maksimoska, Jasna, Villanueva, Jessie, Winkler, Jeffrey D., & Marmorstein, Ronen. Chemically Linked Vemurafenib Inhibitors Promote an Inactive BRAFV600E Conformation. United States. https://doi.org/10.1021/acschembio.6b00529
Grasso, Michael, Estrada, Michelle A., Ventocilla, Christian, Samanta, Minu, Maksimoska, Jasna, Villanueva, Jessie, Winkler, Jeffrey D., and Marmorstein, Ronen. Thu .
"Chemically Linked Vemurafenib Inhibitors Promote an Inactive BRAFV600E Conformation". United States. https://doi.org/10.1021/acschembio.6b00529. https://www.osti.gov/servlets/purl/1331701.
@article{osti_1331701,
title = {Chemically Linked Vemurafenib Inhibitors Promote an Inactive BRAFV600E Conformation},
author = {Grasso, Michael and Estrada, Michelle A. and Ventocilla, Christian and Samanta, Minu and Maksimoska, Jasna and Villanueva, Jessie and Winkler, Jeffrey D. and Marmorstein, Ronen},
abstractNote = {The BRAF kinase, within the mitogen activated protein kinase (MAPK) signaling pathway, harbors activating mutations in about half of melanomas and to a significant extent in many other cancers. A single valine to glutamic acid substitution at residue 600 (BRAFV600E) accounts for about 90% of these activating mutations. While BRAFV600E-selective small molecule inhibitors, such as debrafenib and vemurafenib, have shown therapeutic benefit, almost all patients develop resistance. Resistance often arises through reactivation of the MAPK pathway, typically through mutation of upstream RAS, downstream MEK, or splicing variants. RAF kinases signal as homo- and heterodimers, and another complication associated with small molecule BRAFV600E inhibition is drug-induced allosteric activation of a wild-type RAF subunit (BRAF or CRAF) of the kinase dimer, a process called “transactivation” or “paradoxical activation.” Here, we used BRAFV600E and vemurafenib as a model system to develop chemically linked kinase inhibitors to lock RAF dimers in an inactive conformation that cannot undergo transactivation. This structure-based design effort resulted in the development of Vem-BisAmide-2, a compound containing two vemurafenib molecules connected by a bis amide linker. We show that Vem-BisAmide-2 has comparable inhibitory potency as vemurafenib to BRAFV600E both in vitro and in cells but promotes an inactive dimeric BRAFV600E conformation unable to undergo transactivation. The crystal structure of a BRAFV600E/Vem-BisAmide-2 complex and associated biochemical studies reveal the molecular basis for how Vem-BisAmide-2 mediates selectivity for an inactive over an active dimeric BRAFV600E conformation. These studies have implications for targeting BRAFV600E/RAF heterodimers and other kinase dimers for therapy.},
doi = {10.1021/acschembio.6b00529},
journal = {ACS Chemical Biology},
number = 10,
volume = 11,
place = {United States},
year = {Thu Aug 25 00:00:00 EDT 2016},
month = {Thu Aug 25 00:00:00 EDT 2016}
}
Web of Science
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