Multimeric Oligonucleotide Linkers for Stable Cellular Delivery
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for synthesizing multimeric oligonucleotides face limitations such as the inability to introduce disulfide linkages while maintaining internal disulfides, random polymerization in asymmetric annealing, and difficulty in forming homo-multimers, leading to inefficient delivery and internalization of oligonucleotides.
Innovation Solution
The synthesis of multimeric oligonucleotides using covalent linkers that join oligonucleotide subunits, including partial oligonucleotides, allows for the formation of double-stranded subunits with complementary strands and cleavable linkers, enhancing stability and delivery efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If disulfide linkers are used to join oligonucleotide subunits, then multimeric oligonucleotides can be formed with enhanced serum half-lives and bioactivities, but it is not possible to maintain an internal disulfide group while simultaneously reducing a terminal disulfide to a thiol for subsequent linking reactions
Solution Approach 1:
The patent segments the oligonucleotide structure into subunits joined by disulfide linkers, enabling modular assembly while maintaining stability. The segmentation allows different regions (internal vs terminal disulfides) to have different functions - internal disulfides maintain structural integrity for serum half-life extension, while terminal disulfides can be reduced for further linking reactions.
Solution Approach 2:
The patent applies local quality by differentiating the chemical properties of disulfide groups at different positions. Internal disulfide groups maintain their oxidized state for structural stability and serum half-life extension, while terminal disulfide groups are reduced to thiols to enable subsequent linking reactions. This spatial differentiation of chemical states resolves the contradiction between stability and reactivity.
2Manufacturing precision
If asymmetric annealing is used to synthesize multimeric oligonucleotides, then hetero-multimers can be formed with controlled structure, but the method is difficult to apply to homo-multimers due to random polymerization
Solution Approach 1:
The patent employs asymmetric annealing where a single-stranded oligonucleotide bonded via a linker to another oligonucleotide is annealed to a complementary single-stranded oligonucleotide. This asymmetric approach provides controlled structure for hetero-multimers while the modular design with standardized linkers enables extension to homo-multimers, resolving the limitation of random polymerization.
Solution Approach 2:
The patent uses linker molecules as intermediaries between oligonucleotide subunits. These linkers facilitate controlled assembly by providing defined attachment points and spacing, preventing random polymerization while enabling the formation of both hetero-multimers and homo-multimers with predictable structures.
3Duration of action of stationary object
If multimers of oligonucleotides are prepared to counter kidney excretion, then serum half-life is extended, but the delivery and internalization into target cells in sufficient quantities remains limited
Solution Approach 1:
The patent creates composite structures by conjugating oligonucleotide multimers to cell-penetrating peptides or other delivery vehicles. This composite approach combines the serum half-life extension benefits of multimerization with the cell delivery capabilities of the attached peptide or vehicle, simultaneously addressing both stability and delivery efficiency.
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 method improves the serum half-life and bioactivity of multimeric oligonucleotides, enabling effective delivery and internalization into target cells, addressing the limitations of existing synthesis techniques.
Implementation Method 1
asymmetric annealing whereby a single-stranded oligonucleotide bonded via a linker to another oligonucleotide is annealed to a complementary single-stranded oligonucleotide
Implementation Method 2
two partial oligonucleotide strands are complementary, and together are anneal to form a double-stranded oligonucleotide subunit
Implementation Method 3
oligonucleotide subunits joined together by covalent linkers
Implementation Method 4
disulfide-based linkers which are cleaved by the reductive environment inside the cell
Implementation Method 5
short sequences of single-stranded unprotected nucleotides such as dTdTdTdT and dCdA, which are cleaved by intracellular nucleases
Data Source
AI summary
The present disclosure relates to multimeric oligonucleotides comprising subunits, each of the subunits independently comprises a single-stranded or double-stranded oligonucleotide. Each of the subunits is joined to another subunit by a covalent linker, and at least one subunit comprises at least one partial single-stranded oligonucleotide. The present disclosure also relates to methods of synthesizing the multimeric oligonucleotides and the methods of using the multimeric oligonucleotides disclosed herein.