Reversible Thioether Bonding via Electrophilic Leaving Groups
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for conjugating molecules using maleimide cross-linkers are limited by the irreversibility of the thioether bond, making it difficult to regenerate native thiol-containing reagents and limiting their utility in protein purification, proteomic analysis, and drug delivery applications.
Innovation Solution
Incorporating an electrophilic leaving group onto the C═C double bond of 1,2-dicarbonyl ethene cross-linking reagents allows for reversible thioether bond formation, enabling controlled cleavage and easier addition of further functional moieties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If maleimide cross-linkers are used to conjugate molecules via thioether bond formation, then conjugation is achieved with high selectivity and stability, but the bond is irreversible making it difficult to regenerate native thiol-containing reagents
Solution Approach 1:
The patent modifies the maleimide cross-linker by incorporating an electrophilic leaving group (such as halogen atoms at positions 2 or 3 of the maleimide ring) to change the chemical parameters of the cross-linking system. This enables the thioether bond to become reversible through nucleophilic attack, allowing regeneration of the thiol group while maintaining conjugation stability during the conjugation process
Solution Approach 2:
The patent introduces an electrophilic leaving group as an intermediary element in the maleimide cross-linker structure. This leaving group acts as a temporary participant that enables reversible bond formation - it is displaced during conjugation but can be removed later to regenerate the original thiol-containing reagent, thus serving as a mediator between irreversible and reversible bonding requirements
2Ease of manufacture
If conventional maleimide cross-linkers are used, then conjugation proceeds with standard reagents, but further functionalization is limited due to the irreversible nature of the bond and lack of reactive sites
Solution Approach 1:
The patent creates a multi-functional cross-linking system where the maleimide ring with electrophilic leaving groups serves multiple purposes: it enables reversible thiol conjugation, provides reactive sites for further functionalization through nucleophilic attack, and maintains conjugation stability. This single modified cross-linker replaces multiple separate reagents and steps, achieving both ease of manufacture and enhanced versatility
Solution Approach 2:
The patent introduces dynamic reversibility to the previously static irreversible bond. The electrophilic leaving groups enable the bond to transition between bound and unbound states, allowing the system to adapt from a fixed conjugation state to a dynamic system that can be modified further, thus enhancing functionalization capability while maintaining process simplicity
3Stability of the object's composition
If thiol-containing reagents are permanently modified by maleimide cross-linking, then conjugate stability is achieved, but the native reagent cannot be regenerated for repeated use
Solution Approach 1:
The patent enables the electrophilic leaving group to be discarded during conjugation (displaced by the thiol nucleophile) but then recovered or replaced later to regenerate the native thiol-containing reagent. This allows the conjugate to remain stable during application while the original reagent can be recovered and reused, preventing permanent loss of valuable thiol reagents
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 allows for rapid and high-yield conjugation with stoichiometric amounts of cross-linker, facilitating reversible bonding and enhanced functionalization of conjugate molecules, applicable across various biotechnological methods.
Implementation Method 1
The thiol group in a thiol-containing functional molecule ('R—SH') reacts with maleimide to produce a thioether linkage
Data Source
AI summary
The present invention relates to the use of a compound containing a moiety of formula (I) as a reagent for linking a compound of formula R1—H which comprises a first functional moiety of formula F1 to a second functional moiety of formula F2wherein X, X′, Y, R1, F1 and F2 are as defined herein. The present invention also provides related processes and products. The present invention is useful for creating functional conjugate compounds, and specifically conjugates in which at least one of the constituent molecules carries a thiol group.