Bacterially Secretable Single-Chain Tandem Macrocyclic Peptides for High Affinity and Inhibitory Activity

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Protein-protein interactions (PPIs) are crucial in cellular processes and represent therapeutic targets.
• Macrocyclic peptides are promising for PPI inhibition due to their binding surface area.
• Targeting vascular endothelial growth factor receptor 2 (VEGFR2) is relevant for various diseases.

Purpose of the Study:

• To design and develop novel single-chain tandem macrocyclic peptides (STaMPtides) for inhibiting VEGFR2.
• To evaluate the inhibitory activity of STaMPtides against VEGFR2.
• To explore the potential of STaMPtides for targeting other extracellular factors.

Main Methods:

• Artificial design of STaMPtides by linking two different VEGFR2-binding macrocyclic peptides.
• Production of STaMPtides using Corynebacterium glutamicum secretion.
• Optimization of peptide linker lengths for enhanced activity.
• Assay of VEGFR2 inhibitory activity and comparison with monomeric peptides.

Main Results:

• Developed STaMPtides capable of inhibiting VEGFR2.
• Optimized STaMPtides demonstrated >1000-fold increased inhibitory activity compared to parental monomeric peptides.
• The enhanced activity is potentially attributed to the avidity effect of heterodimerization.

Conclusions:

• STaMPtides represent a potent new class of molecules for PPI inhibition.
• This approach offers a promising strategy for developing therapeutics targeting VEGFR2.
• The STaMPtide technology may be applicable for inhibiting other growth factors and cytokines.