5′-Capped Oligonucleotide Synthesis With Hydrophobic Tag Purification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for producing mRNA lack effective in scale-up and high purity, stable 5'-capped mRNA are not effective in addressing the 5'-capped mRNA are not effective in addressing the 5'-capped mRNA are not effective in addressing the 5'-capped mRNA are not effective in solving the 5'-capped mRNA synthesis.
Innovation Solution
A method involving reacting a 5'-phosphate-oligonucleotide with a modified Im-m7GDP, which includes a cleavable or non-cleavable hydrophobic moiety, followed by purification to produce highly pure, stable 5'-capped oligonucleotides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional enzymatic methods are used to prepare 5'-capped mRNA, then the mRNA can be produced with native cap structures, but the process is costly and not amenable to scale-up
Solution Approach 1:
The patent replaces enzymatic methods (biological system) with chemical synthesis methods (chemical system) for cap structure formation. This substitution enables non-enzymatic cap analog incorporation during oligonucleotide synthesis, making the process more amenable to scale-up and cost-effective while maintaining product quality
Solution Approach 2:
The patent modifies chemical reaction parameters and conditions to achieve efficient non-enzymatic capping. By optimizing coupling reagents, reaction times, and purification conditions, the method achieves high purity levels and scalability without requiring enzymatic processes
2Productivity
If non-enzymatic chemical synthesis methods are used to make mRNA, then the process can be more cost effective and amenable to scale-up, but achieving high purity and stability of 5'-capped mRNA with 40 or longer bases is challenging
Solution Approach 1:
The patent employs hydrophobic tags as intermediary groups that facilitate purification. These tags are temporarily attached during synthesis to enable selective purification of capped oligonucleotides from uncapped byproducts, then removed to yield high purity final products. This intermediary approach solves the purification challenge in non-enzymatic synthesis
Solution Approach 2:
The patent segments the synthesis process into distinct stages: cap analog incorporation, hydrophobic tag attachment, purification, and tag removal. This segmentation allows each step to be optimized independently, ensuring high purity and stability while maintaining cost-effectiveness and scalability
3Adaptability or versatility
If standard cap structures are used, then the mRNA can be produced, but translation efficiency and immunogenicity control are limited
Solution Approach 1:
The patent applies local quality modification by introducing site-specific modifications at the cap structure and nearby regions. By controlling the chemical environment and composition locally at the 5' end, the method optimizes translation efficiency while modulating immunogenicity responses, achieving tailored mRNA performance
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 enables the production of highly pure, stable, and translatable 5'-capped oligonucleotides with site-specific modifications, enhancing translation activity and reducing immunogenicity.
Implementation Method 1
reacting a 5'-phosphate-oligonucleotide with a modified Im-m7GDP
Implementation Method 2
the hydrophobic group can be removed following purification of the product generated from the reaction
Data Source
AI summary
Described herein are highly pure, chemically synthesized, stabilized, 5′-capped oligonucleotides. Additionally, described herein are methods for making and using said oligonucleotides.