RNA Monomer Synthesis via Lipase Selective Esterification

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Solution Overview

Problem

Conventional methods for producing RNA monomers are inefficient, requiring numerous steps and excessive amounts of reagents, leading to high production costs and waste, and are not suitable for mass production due to the need for selective protection of hydroxy groups and excessive monomer usage.

Innovation Solution

A method involving the use of a monomer with a 2'-position or 3'-position protected by an alkanoyl group, where the 2'-hydroxy or 3'-hydroxy group is selectively esterified using a lipase, reducing the number of steps and reagents needed, and allowing for efficient production with approximately stoichiometric amounts, thereby lowering production costs and improving yield.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional methods are used to selectively protect hydroxy groups, then selectivity is achieved, but the number of synthesis steps increases to at least seven steps

Engineering Contradiction:
Improveselectivity of hydroxy group protectionVSAvoidnumber of synthesis steps
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention applies preliminary action by pre-protecting the 5'-hydroxy group before phosphonate esterification. This preliminary protection step allows subsequent selective esterification at the 2'- or 3'-position without needing additional protective groups, reducing the total number of steps from seven to five while maintaining selectivity.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The invention uses local quality by employing different protective strategies for different hydroxy groups. The 5'-hydroxy group is protected with a specific protecting group, while the 2'- or 3'-hydroxy group is selectively esterified with a phosphonate group. This localized differentiation eliminates the need for cyclic protective groups and multiple deprotection steps.

Inventive Principle:
Principle #3Local quality

2Manufacturing precision

If conventional methods are used for RNA monomer production, then selectivity is maintained, but excessive amounts of reagents and monomers are required leading to high costs

Engineering Contradiction:
Improveselectivity of phosphodiester bond formationVSAvoidamount of reagents and monomers
Core Design Contradiction:
Manufacturing precisionVSQuantity of substance

Solution Approach 1:

The invention extracts the unnecessary excess reagent requirement by implementing a streamlined synthesis pathway. By pre-protecting the 5'-hydroxy group and selectively esterifying only one other hydroxy group, the method eliminates the need for excess monomers and reagents typically required to drive equilibrium reactions, enabling cost-effective mass production while maintaining selectivity.

Inventive Principle:
Principle #2Taking out (Extraction)

3Reliability

If conventional methods are used, then RNA synthesis can proceed, but production cost becomes prohibitively high due to multiple steps and excessive reagent usage

Engineering Contradiction:
ImproveRNA synthesis capabilityVSAvoidproduction cost
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The invention applies continuity of useful action by creating a streamlined, continuous synthesis pathway. The pre-protected ribonucleoside undergoes selective phosphonate esterification followed directly by condensation with another ribonucleoside, eliminating intermediate purification and reagent removal steps. This continuous process maintains RNA synthesis reliability while dramatically reducing production costs and enabling scalable manufacturing.

Inventive Principle:
Principle #20Continuity of useful action

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 approach significantly reduces the number of steps and reagents required, enabling cost-effective and efficient production of RNA monomers and RNA, making it industrially useful for mass production while maintaining high selectivity and yield.

Implementation Method 1

the 2'-hydroxy or 3'-hydroxy group is selectively esterified using a lipase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP2878604B1Monomer for synthesis of RNA, method for producing same, and method for producing RNA
Publication Date: 2018.08.29 NATIONAL UNIVERSITY CORPORATION KOCHI UNIVERSITY
  • EP2878604B1 patent drawingFigure 1
  • EP2878604B1 patent drawingFigure 2
  • EP2878604B1 patent drawingFigure 3

AI summary

The objective of the present invention is to provide a monomer for RNA synthesis which can be efficiently produced and therefore by which the producing cost of RNA can be remarkably decreased, and a method for efficiently producing the monomer in a small number of steps. In addition, the objective of the present invention is also to provide a method by which RNA can be efficiently produced even when a approximately stoichiometry amount of the monomer for RNA synthesis is used. The monomer for RNA synthesis according to the present invention is represented by the following formula (I) or (I'): wherein R1 is a protective group of the hydroxy group and R2 is an alkyl group or the like.