Chimeric Peptide Conjugates for PMO Nuclear Delivery
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Solution Overview
Problem
Existing antisense technologies face challenges in efficiently delivering phosphorodiamidate morpholino oligonucleotides (PMOs) to the cytosol and nucleus due to their large size and neutral charge, limiting their therapeutic efficacy, particularly in conditions like Duchenne muscular dystrophy.
Innovation Solution
Development of chimeric peptide-oligonucleotide conjugates comprising covalently linked cell-penetrating peptides (CPPs), including amphipathic and oligoarginine peptides, to enhance PMO delivery by promoting cellular uptake through multiple endocytic mechanisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If phosphorodiamidate morpholino oligonucleotides (PMOs) are used for antisense delivery, then gene expression modulation is achieved, but cellular uptake and nuclear localization are limited due to large size and neutral charge
Solution Approach 1:
The patent creates chimeric peptides by combining cell-penetrating peptide sequences with PMO molecules, forming a composite structure that leverages the gene-targeting capability of PMOs and the cell-penetrating ability of the peptide component. This composite approach resolves the contradiction by integrating two functional elements into a single delivery vehicle that overcomes the size and charge limitations of PMOs alone.
Solution Approach 2:
The cell-penetrating peptide acts as an intermediary that facilitates the entry of the PMO into cells and its subsequent localization to the nucleus. The peptide component mediates the interaction between the PMO and cellular structures, enabling efficient uptake and delivery to the target site without requiring the PMO to overcome its inherent delivery barriers alone.
2Measurement precision
If PMO size and neutral charge are maintained for specificity, then gene targeting precision is preserved, but therapeutic efficacy is limited due to poor cellular penetration
Solution Approach 1:
The chimeric peptide is constructed by segmenting the delivery function from the targeting function. The cell-penetrating peptide segment handles cellular uptake and nuclear localization, while the PMO segment maintains its sequence-specific gene targeting capability. This segmentation allows each component to optimize its specialized function without compromising the other.
Solution Approach 2:
The chimeric peptide structure provides multi-functionality by combining the gene-targeting specificity of PMOs with the cell-penetrating and nuclear-localizing capabilities of the peptide component. This universal design enables a single molecule to perform multiple functions: specific gene recognition, efficient cellular entry, and nuclear delivery, thereby resolving the contradiction between specificity and efficacy.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The chimeric peptide-oligonucleotide conjugates significantly improve PMO delivery to the nucleus, enhancing therapeutic efficacy by increasing cellular uptake and nuclear localization.
Implementation Method 1
enhance PMO delivery by promoting cellular uptake through multiple endocytic mechanisms
Data Source
AI summary
Provided herein are oligonucleotides, chimeric peptides, and peptide-oligonucleotide-conjugates, the chimeric peptides and peptide-oligonucleotide-conjugates comprising at least two cell-penetrating peptides, wherein at least one of the cell-penetrating peptides is an amphipathic peptide and at least one of the cell-penetrating peptides is an oligoarginine peptide. The oligoarginine peptide comprises the sequence [(RYzR)x](SEQ ID NOs: 15-18), wherein R is arginine, Y is independently selected from aminohexanoic acid (X) or β-alanine (B). Also provided herein are methods of treating a muscle disease, a viral infection, or a bacterial infection in a subject in need thereof, comprising administering to the subject oligonucleotides, chimeric peptides, and peptide-oligonucleotide-conjugates comprising at least two cell-penetrating peptides, wherein at least one of the cell-penetrating peptides is an amphipathic peptide and at least one of the cell-penetrating peptides is an oligoarginine peptide described herein.