Understanding Glucagon Aggregation: In Silico Insights and Experimental Validation.

Area of Science:

• Biochemistry
• Computational Biology
• Pharmaceutical Sciences

Background:

• Peptide aggregation is a major hurdle in developing biopharmaceuticals and understanding neurodegenerative diseases.
• Glucagon is a peptide known for its high propensity to aggregate, posing challenges in its therapeutic applications.

Purpose of the Study:

• To investigate the mechanisms of glucagon aggregation.
• To identify effective countermeasures against glucagon aggregation using computational and experimental approaches.

Main Methods:

• In silico computational simulations were employed to model the interactions of glucagon with potential stabilizing agents.
• Experimental validation was performed using microfluidic modulation spectroscopy (MMS) to confirm simulation findings.

Main Results:

• Lactose was found to stabilize glucagon by increasing its alpha-helical content.
• 2-hydroxypropyl-β-cyclodextrin (2-HPβCD) was shown to disrupt protein-protein interactions, effectively preserving glucagon's monomeric form.
• MMS experiments confirmed that 2-HPβCD maintains glucagon's structural stability, validating its anti-aggregation properties.

Conclusions:

• 2-HPβCD is a promising excipient for preventing glucagon aggregation in therapeutic formulations.
• The study introduces a novel approach combining computational and experimental methods for studying peptide aggregation.
• Findings offer new strategies for mitigating aggregation issues in the development of therapeutic peptides.