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Conditional Cell Penetration of Masked CPPs by an ADEPT-like Approach.

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Summary

Researchers developed a method to improve cell-penetrating peptide (CPP) drug delivery specificity. By masking CPPs with enzyme-cleavable groups and using specific enzymes, they achieved targeted delivery and selective cell killing, reducing off-target effects.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Drug Delivery Systems

Background:

  • Cell-penetrating peptides (CPPs) facilitate intracellular cargo delivery but lack cell-type specificity, causing off-target effects.
  • Developing targeted CPPs is crucial for safe and effective drug delivery applications.

Purpose of the Study:

  • To engineer a conditional and selective activation of cellular uptake using a model CPP, the L17E peptide.
  • To demonstrate enzyme-mediated deprotection for controlled CPP activation and targeted delivery.

Main Methods:

  • Lysine residues of the L17E peptide were masked with enzyme-cleavable acetyl protecting groups.
  • Cellular uptake of a conjugated fluorophore (TAMRA) was assessed with masked and unmasked peptides.
  • In vitro deacetylation was performed using NAD-dependent sirtuin 2 (SirT2) deacetylase.
  • Antibody-enzyme conjugates (trastuzumab-SirT2, anti-B7H3-SirT2) were created for targeted delivery of a cytotoxin.

Main Results:

  • Masking the L17E peptide significantly reduced fluorophore delivery to HeLa cells.
  • SirT2 deacetylase restored cellular uptake of the masked L17E peptide.
  • Antibody-enzyme conjugates enabled conditional and selective delivery of a cryptophycin cytotoxin.
  • Selective cell killing was observed with antibody-enzyme conjugates, despite some residual cytotoxicity of the masked peptide.

Conclusions:

  • Enzyme-cleavable masking of CPPs offers a strategy for conditional and selective intracellular delivery.
  • Antibody-targeted enzyme-mediated deprotection can achieve specific drug delivery and cell killing.
  • This approach holds promise for enhancing the safety and efficacy of CPP-based therapeutics.