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Design, Synthesis, and Evaluation of Highly Potent FAK-Targeting PROTACs
Hongying Gao1,2, Yue Wu1, Yonghui Sun1
1MOE Key Laboratory of Protein Sciences, School of Pharmaceutical Sciences, MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, P. R. China.
Abstract:
Focal adhesion kinase (FAK), a cytoplasmic protein tyrosine kinase, exerts kinase-dependent enzymatic functions and kinase-independent scaffolding functions, both of which are crucial in cancer development, early embryonic development, and reproduction. However, previous efforts for FAK blocking mainly focus on kinase inhibitors. Proteolysis targeting chimeras (PROTACs) are heterobifunctional molecules that allow direct post-translational knockdown of proteins via ubiquitination of a target protein by E3 ubiquitin ligase and subsequent proteasomal degradation. Here, we designed and synthesized a FAK PROTAC library with FAK inhibitor (PF562271 or VS6063) and CRBN E3 ligand. A novel FAK-targeting PROTAC, FC-11, showed a rapid and reversible FAK degradation with a picomolar of DC50 in various cell lines in vitro, which imply that FAK-PROTACs could be useful as expand tools for studying functions of FAK in biological system and as potential therapeutic agents.
Insights
New proteolysis targeting chimeras (PROTACs) rapidly degrade focal adhesion kinase (FAK). These FAK PROTACs offer novel tools for biological research and potential therapeutic applications in cancer and reproduction.
Area of Science:
- Molecular Biology
- Biochemistry
- Cancer Research
Background:
- Focal adhesion kinase (FAK) is a cytoplasmic protein tyrosine kinase with critical roles in cancer, embryonic development, and reproduction.
- Existing FAK inhibition strategies primarily utilize kinase inhibitors, leaving other functional aspects unaddressed.
Purpose of the Study:
- To design and synthesize a library of FAK-targeting Proteolysis Targeting Chimeras (PROTACs).
- To evaluate the efficacy of these PROTACs in achieving FAK protein knockdown via targeted protein degradation.
Main Methods:
- Synthesis of a FAK PROTAC library incorporating FAK inhibitors (PF562271 or VS6063) and a CRBN E3 ligase ligand.
- In vitro assessment of FAK degradation in various cell lines using the developed PROTACs.
Main Results:
- A novel FAK-targeting PROTAC, designated FC-11, demonstrated rapid and reversible FAK degradation.
- FC-11 achieved degradation with a sub-nanomolar degradation capacity (DC50) in tested cell lines.
Conclusions:
- FAK PROTACs represent a promising new class of molecules for targeted protein knockdown.
- These FAK PROTACs can serve as valuable tools for investigating FAK functions and as potential therapeutic agents.
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