Utilizing a novel ADAM10 inhibitory peptide to enhance recovery and reduce IL-1-driven inflammation after myocardial infarction

Background and Purpose. Inflammation plays a pivotal role in the progression of both acute and chronic diseases. Targeting ADAM proteases, especially ADAM10, shows promise in modulating complex inflammatory processes such as those occuring after myocardial infarction (MI). However, developing selective ADAM10 inhibitors remains challenging due to the high cross-reactivity with other metalloproteases. Artificial intelligence-driven tools like AlphaFold3 have the potential to streamline and accelerate inhibitor development, improving efficiency from target identification to validation. Protein-based inhibitors, such as engineered ADAM10 prodomains, offer a potential route for achieving greater selectivity and effectiveness. In this study, we utilized machine learning-based modeling combined with in vitro and in vivo experimental studies to identify and validate a novel peptide-based ADAM10 inhibitor derived from the ADAM10 prodomain and assess its selectivity, efficacy, and toxicity.

Methods and Results. We applied AlphaFold3 predictions to model the structures of ADAM10 and nine additional metalloproteases typically affected by common ADAM10 inhibitors. This machine learning-based approach identified the ADAM10 prodomain as a highly selective framework for designing a peptide-based inhibitor. Subsequent AlphaFold3 modeling and FRET-based inhibition assays confirmed that an engineered ADAM10 prodomain selectively targets ADAM10, sparing other metalloproteases. In a mouse model of acute myocardial infarction (MI), administering the novel ADAM10 inhibitor at 4 and 48 hours post-ligation of the left anterior descending (LAD) artery preserved cardiac function by reducing scar size. Peptide-based ADAM10 inhibition reduced CX3CL1 shedding, leading to lower neutrophil infiltration and reduced IL-1β-driven inflammation in the heart. Toxicological assessments in naïve mice revealed no short-term toxicity in cardiac, hepatic, renal and lung tissues with the prodomain-based ADAM10 inhibitors.

Conclusion. Our findings demonstrate that an engineered ADAM10 prodomain, serving as a novel selective ADAM10 inhibitor, effectively preserves cardiac function following MI and holds significant potential as a safe and effective therapeutic option for myocardial infarction and other ADAM10- and inflammation-related diseases.