Stable 4-isoxazoline Conjugates via Nitrone Substituent Design

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Solution Overview

Problem

The instability of 4-isoxazoline rings formed through strain-promoted alkyne-nitrone cycloaddition (SPANC) reactions in bioconjugates leads to premature breakdown, causing unintended release of cytotoxic payloads and reducing the therapeutic efficacy and safety of antibody-drug conjugates.

Innovation Solution

Development of nitrones with specific substituents that capture imine intermediates, preventing rearrangement and maintaining the covalent connection between biomolecules and payloads, thereby stabilizing the bioconjugates and ensuring controlled release of the payload at the target site.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If strain-promoted alkyne-nitrone cycloaddition (SPANC) is used to form 4-isoxazoline rings in bioconjugates, then covalent connection between biomolecules and payloads is achieved, but the 4-isoxazoline rings are unstable and undergo rearrangement leading to premature payload release

Engineering Contradiction:
Improvestability of 4-isoxazoline ringVSAvoidstructural integrity of bioconjugate
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent introduces a stabilizing substituent on the nitrogen atom of the nitrone that acts as an intermediary to capture the imine intermediate during rearrangement. This substituent (such as a carbonyl group, heteroatom, or electron-withdrawing group) intercepts the reactive imine species, preventing the decomposition pathway and stabilizing the 4-isoxazoline ring structure throughout circulation and cellular uptake.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Reliability

If stable 4-isoxazoline rings are achieved through nitrone substituents, then premature payload release is reduced, but the complexity of nitrone design and synthesis increases

Engineering Contradiction:
Improvestability of bioconjugateVSAvoidcomplexity of nitrone structure
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent systematically varies the parameters of the nitrone substituent (electron-withdrawing capability, atomic number, position) to optimize stability. By changing these chemical parameters, the patent identifies that specific classes of substituents (carbonyl, heteroatom, electron-withdrawing groups) provide optimal stabilization while maintaining reasonable structural complexity and synthetic accessibility.

Inventive Principle:
Principle #35Parameter changes

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 improved stability of bioconjugates results in enhanced shelf-life, increased tolerability, and improved therapeutic index by reducing unintended payload release and ensuring effective delivery of cytotoxins to cancer cells.

Implementation Method 1

The instability of 4-isoxazoline rings formed through strain-promoted alkyne-nitrone cycloaddition (SPANC) reactions in bioconjugates

Methodology Applied
Scientific EffectStrain-promoted alkyne-nitrone cycloaddition (SPANC): Chemical Bonding

Data Source

PatentEP4450489A1Stable 4-isoxazoline conjugates
Publication Date: 2024.10.23 SYNAFFIX BV
  • EP4450489A1 patent drawingFigure 1~2
  • EP4450489A1 patent drawingFigure 3
  • EP4450489A1 patent drawingFigure 4a~4t

AI summary

The invention concerns nitrones that are capable of forming 4-isoxazoline compounds by strain-promoted alkyne-nitrone cycloaddition (SPANC), which have improved stability. The 4-isoxazoline compounds do not disintegrate by rearrangement into two molecules. As such, the nitrones according to the present invention are suitable for preparing bioconjugates by SPANC reaction. The invention further concerns the thus obtained bioconjugates, as well as the application thereof in targeting cells and treating cancer.