Enzyme

In vivo

Vemurafenib

PTEN

Melanoma

Biology

Gene

Mutant

Chemistry

Protein degradation

Signal transduction

Proteolysis

Cell biology

Metastatic melanoma

Cancer research

Genetics

Biochemistry

Medicine

PI3K/AKT/mTOR pathway

Nature Communications

Over 300 BRAF missense mutations have been identified in patients, yet currently approved drugs target V600 mutants alone. Moreover, acquired resistance inevitably emerges, primarily due to RAF lesions that prevent inhibition of BRAF V600 with current treatments. Therefore, there is a need for new therapies that target other mechanisms of activated BRAF. In this study, we use the Proteolysis Targeting Chimera (PROTAC) technology, which promotes ubiquitination and degradation of neo-substrates, to address the limitations of BRAF inhibitor-based therapies. Using vemurafenib-based PROTACs, we achieve low nanomolar degradation of all classes of BRAF mutants, but spare degradation of WT RAF family members. Our lead PROTAC outperforms vemurafenib in inhibiting cancer cell growth and shows in vivo efficacy in a Class 2 BRAF xenograft model. Mechanistic studies reveal that BRAF WT is spared due to weak ternary complex formation in cells owing to its quiescent inactivated conformation, and activation of BRAF WT sensitizes it to degradation. This study highlights the degree of selectivity achievable with degradation-based approaches by targeting mutant BRAF-driven cancers while sparing BRAF WT , providing an anti-tumor drug modality that expands the therapeutic window.

Mutant-selective degradation by BRAF-targeting PROTACs