Gem-difluorinated C-isopropylgalactoside derivatives for stable gene transcription induction
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Solution Overview
Problem
Current inducers like IPTG for recombinant protein production are hydrolysable, requiring low temperature storage and frequent addition due to decomposition, which complicates experimental procedures and stability.
Innovation Solution
Development of gem-difluorinated C-isopropylgalactoside derivatives that mimic allolactose, featuring a non-hydrolysable CF2 group, providing stability at room temperature and maintaining constant concentration over time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If IPTG is used as an inducer, then gene transcription can be induced, but the compound decomposes over time requiring low temperature storage and frequent addition
Solution Approach 1:
The patent modifies the chemical structure of the inducer by replacing the hydrolysable glycosidic bond with a non-hydrolysable difluoromethylene bridge (CF2), changing the chemical stability parameter while maintaining biological activity. This structural parameter change eliminates decomposition over time, allowing room temperature storage without losing effectiveness.
Solution Approach 2:
The invention creates a hybrid molecular structure combining the galactose moiety (for biological recognition) with a difluoromethylene bridge (for chemical stability) and an alkyl group (for non-hydrolysable character). This composite structure integrates the beneficial properties of different chemical components to achieve both stability and functionality.
2Reliability
If allolactose is used as an inducer, then it can trigger transcription of the lac operon, but it is hydrolysable and decomposes in solution
Solution Approach 1:
The patent extracts the essential functional component (the galactose moiety that binds to the repressor protein) from the hydrolysable allolactose molecule and combines it with a stable difluoromethylene bridge, eliminating the hydrolysable glycosidic bond while preserving the transcription induction capability.
Solution Approach 2:
The invention creates a molecular mimic or copy of allolactose that replicates its biological function (binding to LacI repressor and inducing transcription) but uses a chemically stable difluoromethylene bridge instead of the unstable glycosidic bond, thereby copying the function while improving stability.
3Stability of the object's composition
If the glycoside sulfur bond of IPTG is replaced by a methylene group to create IBCG, then stability improves, but structural modification may affect effectiveness
Solution Approach 1:
The patent applies local quality modification by placing fluorine atoms specifically at the anomeric position (C1) of the galactose ring, creating a localized difluoromethylene bridge. This localized structural change provides non-hydrolysable stability exactly where the glycosidic bond would be, while minimizing overall structural modification to preserve biological effectiveness.
Data Source
AI summary
The present invention concerns compounds of the following formula (I) or a pharmaceutically acceptable salt thereof, a stereoisomer or a mixture of stereoisomers in any proportion, in particular a mixture of enantiomers, and particularly a racemate mixture, in which R represents H, OH or OR19, as well as processes for preparing these compounds and their use as inducer for the transcription of genes under control of the lac promoter.