Oligonucleotide Synthesis Using Highly Dispersible Liquid-Phase Support
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Solution Overview
Problem
The solid-phase synthesis method for nucleic acids faces challenges such as scalability issues, high costs due to expensive reagents, and lower reaction yields compared to liquid-phase methods, particularly in RNA synthesis.
Innovation Solution
The use of a highly dispersible liquid-phase support (HDLS) bonded to hydrophobic group-bonded nucleosides allows for efficient oligonucleotide synthesis by dissolving these compounds in non-polar solvents and contacting them with an acid/azole complex compound, enabling a flow reaction that improves coupling efficiency and simplifies product separation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If solid-phase synthesis method is used, then product separation and refinement are simplified, but scalability is lost and production capacity is fixed
Solution Approach 1:
The patent inverts the conventional approach by using a liquid-phase support instead of solid-phase, and by attaching the support to the nucleoside rather than having the nucleoside attached to the support. This inversion enables both easy separation (through precipitation of the liquid-phase support) and scalability (through flow reaction capability)
Solution Approach 2:
The patent changes the physical state parameter of the support from solid to liquid, and modifies the solubility characteristics by using a highly dispersible liquid-phase support that can be controlled to aggregate and precipitate. This parameter change enables the system to achieve both ease of separation and scalability
2Extent of automation
If solid-phase synthesis method is used, then automation is facilitated, but reaction yield is lower compared to liquid-phase method
Solution Approach 1:
The patent inverts the conventional solid-phase approach by using a liquid-phase support that can be controlled to aggregate and precipitate. This inversion maintains the ease of automation while achieving higher reaction yields comparable to liquid-phase methods
3Manufacturing precision
If liquid-phase synthesis is used, then reaction yield is improved, but separation and recovery operations become complex
Solution Approach 1:
The patent introduces a highly dispersible liquid-phase support as an intermediary that can be controlled to aggregate and precipitate. This intermediary enables easy separation of the oligonucleotide product while maintaining the high reaction yields of liquid-phase synthesis
4Productivity
If solid-phase synthesis is used for RNA production, then production can be scaled, but reagent cost increases significantly
Solution Approach 1:
The patent inverts the conventional approach by using a liquid-phase support with controlled aggregation, enabling both scalable production and reduced reagent consumption through improved coupling 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
This method enhances the yield of oligonucleotides, including difficult-to-synthesize RNA, while reducing reagent usage and maintaining consistent reaction efficiency, making it more scalable and cost-effective than traditional methods.
Implementation Method 1
a non-polar solvent, and contacting the resulting solution with an acid/azole complex compound
Implementation Method 2
a 5'-hydroxyl group, an amino group of a base, and an amidite monomer having a protected 2'-hydroxyl group, if in RNA, are coupled with a promoter (activator) such as tetrazole compounds and imidazole compounds, and the resulting product is oxidized to form a phosphodiester bond
Implementation Method 3
the resulting product is oxidized to form a phosphodiester bond
Data Source
AI summary
A nucleic acid synthesis method enabling a reaction in a fluid (flow) with a highly dispersible liquid-phase support to improve coupling efficiency is provided. The method for synthesizing an oligonucleotide comprising: sequentially condensing and oxidizing a nucleoside phosphoramidite compound in the presence of an acid/azole complex compound using a starting raw material, i.e., hydrophobic group-bonded nucleoside represented by Formula (1): where R1: an alkylene group having 1 to 12 carbon atoms, R2: an alkylene group having 1 to 22 carbon atoms, R3 and R4 each independently represent an alkyl group having 1 to 22 carbon atoms or the like, R5: a single bond or an alkylene group having 1 to 22 carbon atoms, R6: each independently an alkyl group having 6 to 30 carbon atoms, n represents an integer of 2 to 6, X represents a hydrogen atom, hydroxyl group, or the like, Y: a protecting group deprotectable under an acidic condition, and Z: an adenyl group, a guanyl group, or the like having a polar group optionally protected by a protecting group, wherein a condensation reaction is performed by preliminarily dissolving the hydrophobic group-bonded nucleoside or hydrophobic group-bonded oligonucleotide and the nucleoside phosphoramidite compound in a nonpolar solvent, and contacting the resulting solution with the acid/azole complex compound or a solution containing the complex compound.