Copper-Free Cell Labeling via Strained Cyclooctyne Metabolism
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Solution Overview
Problem
The use of copper catalysts in metabolic labeling methods for adding reporter substances to sugar chains on cell surfaces is toxic, making it difficult to apply these methods to clinical practices.
Innovation Solution
Development of monosaccharide derivatives with a six-membered ring structure that are metabolized to sialic acid, featuring a carbon-carbon double or triple bond, allowing for bioorthogonal reactions without copper catalysts, enabling safe and clinical application.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If CuAAC (copper-catalyzed azide-alkyne cycloaddition) is used to add reporter substances to sugar chains on cell surfaces, then the labeling efficiency and reaction speed are improved, but copper toxicity to living organisms increases, making it unsafe for clinical application
Solution Approach 1:
The patent extracts and removes the copper catalyst from the click reaction system. By using strain-promoted azide-alkyne cycloaddition (SPAAC) with cyclooctyne derivatives, the reaction proceeds without copper catalyst, eliminating copper toxicity while maintaining labeling efficiency for clinical applications
Solution Approach 2:
The patent changes the reaction parameters by using strained cyclooctyne rings with high ring strain energy. This structural modification allows the alkyne to react with azide without copper catalyst, changing the reaction mechanism from copper-catalyzed to strain-promoted, thereby eliminating toxicity
2Reliability
If Ac4ManNAz or Ac4ManNAl is used for metabolic labeling, then sialic acid can be labeled on cell surfaces, but copper catalyst is required for adding reporter substances, which limits clinical application due to toxicity
Solution Approach 1:
The patent introduces cyclooctyne-containing monosaccharide derivatives as intermediaries. These derivatives are metabolized to sialic acid on cell surfaces and present the strained alkyne group, which then serves as a mediator for copper-free click reaction with azide-reporter substances, enabling clinical application
Solution Approach 2:
The patent incorporates the strained cyclooctyne group into the monosaccharide structure beforehand. This preliminary action ensures that when the monosaccharide is metabolized to sialic acid on the cell surface, the reactive alkyne group is already in place, ready for immediate copper-free click reaction without requiring copper catalyst
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
Enables safe and effective cell labeling without the use of copper catalysts, allowing for the application of the cell labeling agent and kit in clinical settings.
Implementation Method 1
metabolized to sialic acid in the sialic acid biosynthetic pathway of cells
Implementation Method 2
allowing for bioorthogonal reactions without copper catalysts
Data Source
AI summary
The cell labeling agent includes a monosaccharide derivatives with a six-membered ring structure that are metabolized to sialic acid in the sialic acid biosynthetic pathway of cells. Among the groups bonded to carbon atoms constituting a six-membered ring in the monosaccharide derivatives, at least one group that does not change, even when metabolized by the sialic acid biosynthetic pathway, includes a ring structure with a carbon-carbon double bond or triple bond.