Phage Display of Two Distinct Warheads to Inhibit Challenging Proteins

Area of Science:

• Biochemistry
• Medicinal Chemistry
• Drug Discovery

Background:

• Peptides offer advantages of both small molecules and antibodies as therapeutic agents.
• High-throughput screening platforms are needed to discover peptide therapeutics with diverse motifs.
• Phage display is a powerful technique for identifying peptide ligands.

Purpose of the Study:

• To develop a novel phage library for discovering peptide therapeutics.
• To incorporate dual, distinct functional motifs onto phage-displayed peptides.
• To demonstrate the utility of this library for identifying enzyme inhibitors.

Main Methods:

• Construction of a novel phage library with two distinct designer groups.
• Sequential chemoselective modification of phage-displayed peptides with reversible covalent warheads targeting cysteine and lysine.
• Screening the double-warhead library against TEV protease.

Main Results:

• Identification of peptide inhibitors with single-digit micromolar potency against TEV protease.
• Structure-activity relationship studies confirmed the contribution of both covalent warheads to inhibition.
• Demonstrated successful dual modification of phage-displayed peptides.

Conclusions:

• The novel double-warhead phage library facilitates the discovery of potent peptide therapeutics.
• This dual modification strategy expands the chemical space accessible via phage display.
• The approach is adaptable for creating diverse phage libraries with various functional motifs.