Mitigating Excessive Mitochondrial Fission Through the Development of a Selective Macrocyclic Inhibitor Targeting...

Area of Science:

• Cardiovascular Biology
• Mitochondrial Dynamics
• Drug Discovery

Background:

• Mitochondrial fragmentation, regulated by Fis1 and Mid51, is linked to cardiovascular disease.
• Targeting protein-protein interactions (PPIs) in mitochondrial dynamics offers therapeutic potential.

Purpose of the Study:

• To identify and optimize inhibitors of the Fis1/Mid51 PPI.
• To evaluate the therapeutic efficacy and safety of these inhibitors in a cellular model of cardiovascular stress.

Main Methods:

• Homologous sequence analysis and structure-guided design were used to identify peptide inhibitors.
• Fluorescence polarization assays assessed binding affinity.
• H9c2 cardiomyocytes were used to evaluate cellular effects under stress.
• In silico ADMET profiling and preliminary in vivo toxicity studies were performed.

Main Results:

• A linear peptide inhibitor (CVP-240) and its macrocyclic derivative (CVP-764) were developed, targeting the Fis1/Mid51 PPI.
• CVP-764 demonstrated high affinity binding to Mid51, selectively disrupted Fis1/Mid51 signaling, and showed nanomolar binding.
• In cardiomyocytes, CVP-764 preserved mitochondrial function, reduced oxidative stress, and enhanced cell viability.
• Macrocyclization improved stability and cell permeability; in silico and in vivo studies suggested a favorable safety profile.

Conclusions:

• CVP-764 is a potent and selective inhibitor of the Fis1/Mid51 interaction.
• The macrocyclic derivative exhibits enhanced stability, cell permeability, and a promising safety profile.
• CVP-764 represents a promising lead compound for therapeutic intervention in pathological mitochondrial fission associated with cardiovascular disease.