Integrating fragment-based screening with targeted protein degradation and genetic rescue to explore eIF4E function

Area of Science:

• Molecular Biology
• Biochemistry
• Drug Discovery

Background:

• Eukaryotic initiation factor 4E (eIF4E) is a critical regulator of oncogene-driven protein synthesis.
• eIF4E is a validated anticancer target due to its role in cancer progression.

Purpose of the Study:

• To identify novel ligands and binding sites on eIF4E using fragment-based screening.
• To develop tool compounds that inhibit eIF4E function and probe its biological role.

Main Methods:

• Fragment library screening against eIF4E.
• Structure-based drug design and medicinal chemistry.
• Biochemical assays to measure eIF4E:eIF4G interaction disruption.
• Cellular assays for translation inhibition and target engagement.
• Targeted protein degradation and genetic rescue experiments.

Main Results:

• Identification of a novel ligand-binding site on eIF4E.
• Development of a low nM tool compound (4) that inhibits eIF4E:eIF4G interaction.
• Demonstration of translation inhibition and eIF4E target engagement in cells.
• Evidence that disrupting both canonical and non-canonical binding sites is necessary for significant cellular effects.

Conclusions:

• Fragment-based drug discovery is effective for targeting challenging proteins like eIF4E.
• Combined approaches of drug discovery, genetic manipulation, and protein degradation can elucidate protein function.
• This study provides a novel chemical probe for eIF4E and insights into its complex regulation in cancer.