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Assembly and Characterization of Polyelectrolyte Complex Micelles
Published on: March 2, 2020
Hydrophobic CPP/HDO conjugates: a new frontier in oligonucleotide-warheaded PROTAC delivery
Miyako Naganuma1,2, Nobumichi Ohoka3, Motoharu Hirano1,2
1Division of Organic Chemistry, National Institute of Health Sciences Kanagawa Japan demizu@nihs.go.jp +81 44 270 6578 +81 44 270 6578.
Abstract:
Proteolysis-targeting chimeras (PROTACs) have emerged as a potent strategy for inducing targeted degradation of proteins, offering promising therapeutic potential to treat diseases such as cancer. However, oligonucleotide-based PROTACs face significant delivery challenges because of their anionic nature and chemical instability. To address these issues, we developed a novel hydrophobic cell-penetrating peptide (CPP) and heteroduplex oligonucleotide (HDO)-conjugated PROTAC, CPP/HDO-PROTAC, to enhance intracellular delivery and degradation efficiency. CPP/HDO-PROTAC was designed to enter the cell through the activity of the conjugated hydrophobic CPP and release decoy oligonucleotide-based PROTACs by RNase H-mediated RNA strand breaks. Our findings demonstrated that CPP/HDO-PROTAC binds to the estrogen receptor α (ERα) with higher affinity than previous constructs, significantly degrades ERα in MCF-7 human breast cancer cells and inhibits cell proliferation at 10 μM. This research highlights the potential of CPP/HDO-PROTAC as a viable method for delivering and activating decoy oligonucleotide-based PROTACs within cells, overcoming the limitations of traditional transfection methods and paving the way for their clinical application.
Insights
A new hydrophobic cell-penetrating peptide and heteroduplex oligonucleotide-conjugated Proteolysis-Targeting Chimera (CPP/HDO-PROTAC) enhances protein degradation. This novel approach overcomes delivery challenges for oligonucleotide-based PROTACs, showing promise for cancer therapy.
Area of Science:
- Biochemistry and Molecular Biology
- Drug Delivery and Nanotechnology
- Oncology and Cancer Therapeutics
Background:
- Proteolysis-targeting chimeras (PROTACs) are a promising therapeutic strategy for targeted protein degradation, particularly in cancer treatment.
- Oligonucleotide-based PROTACs face significant challenges in cellular delivery due to their anionic nature and chemical instability.
- Existing transfection methods for oligonucleotide delivery are often inefficient and can be problematic for clinical applications.
Purpose of the Study:
- To develop a novel delivery system for oligonucleotide-based PROTACs that enhances intracellular delivery and degradation efficiency.
- To overcome the limitations associated with the anionic nature and chemical instability of oligonucleotide-based PROTACs.
- To investigate the efficacy of a new hydrophobic cell-penetrating peptide (CPP) and heteroduplex oligonucleotide (HDO)-conjugated PROTAC (CPP/HDO-PROTAC) for targeted protein degradation.
Main Methods:
- Design and synthesis of a novel CPP/HDO-PROTAC conjugate.
- Evaluation of cellular uptake mechanism mediated by the hydrophobic CPP.
- Assessment of RNase H-mediated release of decoy oligonucleotide-based PROTACs.
- Measurement of estrogen receptor α (ERα) binding affinity and degradation efficiency in MCF-7 breast cancer cells.
- Analysis of cell proliferation inhibition by the CPP/HDO-PROTAC.
Main Results:
- The developed CPP/HDO-PROTAC demonstrated enhanced binding affinity to estrogen receptor α (ERα) compared to previous constructs.
- Significant degradation of ERα was observed in MCF-7 human breast cancer cells treated with CPP/HDO-PROTAC.
- CPP/HDO-PROTAC effectively inhibited cancer cell proliferation at a concentration of 10 μM.
Conclusions:
- CPP/HDO-PROTAC represents a viable strategy for efficient intracellular delivery and activation of decoy oligonucleotide-based PROTACs.
- This novel approach overcomes the limitations of traditional transfection methods for oligonucleotide-based therapeutics.
- The findings support the potential clinical application of CPP/HDO-PROTAC for targeted protein degradation in diseases like cancer.
