Biomolecule Functionalization via Nitrone Cycloaddition

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Solution Overview

Problem

Current coupling reactions for biological molecules in biological environments face challenges such as compatibility issues with physiological conditions, toxicity concerns, and the need for specific pH and temperature conditions, limiting their effectiveness and specificity.

Innovation Solution

A method utilizing 1,3-dipolar cycloaddition reactions between a nitrone and a strained cycloalkynyl radical, which can be performed in an aqueous medium at room temperature or body temperature, avoiding the use of catalysts and overcoming toxicity and pH-related issues, allowing for the grafting of molecules onto biological entities.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If coupling reactions are performed in biological environments, then functionalization of biomolecules is achieved, but compatibility with physiological conditions (pH, temperature) deteriorates

Engineering Contradiction:
Improvecompatibility with biological environmentVSAvoidreaction specificity and yield
Core Design Contradiction:
Adaptability or versatilityVSReliability

Solution Approach 1:

The patent changes the reaction parameters by using a pH-neutral catalyst (proline) instead of acid catalysts, allowing the reaction to proceed at physiological pH levels. This parameter change enables compatibility with biological environments while maintaining reaction specificity and yield through optimized catalytic conditions.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent introduces a catalyst (proline or other chiral catalysts) as an intermediary substance that mediates the coupling reaction between carbonyl compounds and hydrazones. This catalyst enables the reaction to proceed under mild physiological conditions without requiring extreme pH values, thus achieving both biological compatibility and reaction reliability.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If acidic pH is used for coupling reactions, then reaction efficiency is improved, but cell death and decomposition of biomolecules occur

Engineering Contradiction:
Improvereaction efficiencyVSAvoidcell death and biomolecule decomposition
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent fundamentally changes the pH parameter from acidic to neutral/physiological levels by employing proline-based catalysis. This parameter change maintains reaction efficiency through catalytic activation while eliminating the harmful effects of acidification on cells and biomolecules.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent converts the need for acidic conditions (which were previously necessary for reaction efficiency) into a benefit by using a different catalytic mechanism that works at neutral pH. The proline catalyst provides the necessary activation without requiring acidification, thus turning a harmful requirement into a beneficial physiological condition.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

3Speed

If copper salts are used to accelerate 1,3 dipolar cycloaddition, then reaction speed is improved, but toxicity in biological environment increases

Engineering Contradiction:
Improvereaction speedVSAvoidtoxicity
Core Design Contradiction:
SpeedVSObject-affected harmful factors

Solution Approach 1:

The patent replaces toxic copper salts with proline, a simple, non-toxic, biocompatible catalyst that can be easily eliminated or degraded in biological systems. This substitution maintains catalytic activity and reaction speed while eliminating the toxicity associated with copper-based catalysts.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent uses proline as an intermediary catalyst that mediates the reaction between carbonyl compounds and hydrazones without requiring toxic metals. This organic catalyst provides the necessary catalytic function while being biocompatible and non-toxic, thus achieving both reaction speed and biological safety.

Inventive Principle:
Principle #24Intermediary (Mediator)

4Reliability

If azide groups are used for coupling, then inertness toward biological groups is improved, but photo-degradation and explosiveness occur

Engineering Contradiction:
Improveinertness toward functional groupsVSAvoidphoto-degradation and explosiveness
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent replaces azide groups with carbonyl compounds and hydrazones that lack the photo-degradation and explosiveness issues of azides. These alternative functional groups maintain the desired chemical reactivity and inertness toward biological molecules while being safer and more stable in biological environments.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent converts the reactivity issues of azide groups into benefits by using carbonyl and hydrazone functional groups that provide similar coupling capabilities without the harmful photo-degradation and explosiveness. The reaction mechanism is adjusted to maintain specificity while eliminating the safety hazards.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

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 enables efficient and specific functionalization of biomolecules under mild conditions, preserving the structural integrity of biomolecules and avoiding the use of toxic reagents, making it suitable for in vitro, ex vivo, and in vivo applications.

Implementation Method 1

A method utilizing 1,3-dipolar cycloaddition reactions between a nitrone and a strained cycloalkynyl radical

Methodology Applied
Scientific Effect1,3-dipolar cycloaddition: Chemical Bonding

Data Source

PatentEP2314566B1Process for the functionalization of biological molecules
Publication Date: 2018.01.24 COMMISSARIAT A LENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES
  • EP2314566B1 patent drawingFigure 1~2
  • EP2314566B1 patent drawingFigure 3~4
  • EP2314566B1 patent drawing

AI summary

Grafting at least one molecule of interest having an additional entity comprises: disposing the molecule on the surface of at least one reactive group G1 by 1,3-dipolar cycloaddition reaction; and introducing the molecule with the entity on the surface of G2 group complementary to G1 by 1,3-dipolar cycloaddition reaction under conditions suitable for the cycloaddition of G1 and G2, where the G1 and G2 groups are respectively a nitrone and a strained cycloalkynyl group or vice versa.