Reductively Cleavable Peptide Linkers for On-Resin Modification
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Solution Overview
Problem
Current methods for modifying peptides, such as lipidation and cyclization, are cumbersome and often result in false-positive or false-negative results due to the need for orthogonal protection strategies and purification challenges, especially when performed on solid supports.
Innovation Solution
A method utilizing traceless reductively cleavable linker molecules to immobilize peptides on a solid support, allowing for simultaneous modification and purification without orthogonal protection, using a linker molecule of formula X-Tb-Va—U—Y—Z, which is stable under acidic conditions and releases the modified peptide through reduction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If orthogonal protection strategies are used for peptide modification on solid supports, then selective modification can be achieved, but the process becomes cumbersome and requires extra purification steps
Solution Approach 1:
The patent extracts the protection group strategy entirely by using a traceless linker that directly attaches to the peptide without requiring orthogonal protection. The linker is incorporated during standard SPPS and subsequently cleaved under mild reductive conditions, eliminating the need for additional protecting groups and their associated deprotection steps.
Solution Approach 2:
The traceless linker serves multiple functions: it acts as a temporary anchor during SPPS, enables selective modification of the immobilized peptide, and is subsequently removed under mild conditions. This multi-functional approach replaces the need for multiple orthogonal protection strategies, simplifying the overall process.
2Ease of manufacture
If peptides are modified on solid supports using traditional methods, then modification can be performed, but purification becomes difficult or impossible
Solution Approach 1:
The patent performs the modification action while the peptide is still immobilized on the solid support through the traceless linker. This preliminary modification before cleavage allows excess reagents and byproducts to be easily removed by washing, and the modified peptide to be cleaved in high purity without requiring additional purification steps.
3Manufacturing precision
If excess reagent is used during modification on solid support, then complete modification can be achieved, but side reactions increase
Solution Approach 1:
The patent extracts the problematic excess reagent issue by performing modification on immobilized peptides. The solid support acts as a filtering mechanism that retains the modified peptide while allowing excess reagent and byproducts to be washed away, eliminating the need to use precise stoichiometric amounts and reducing side reactions from excess reagent.
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 efficient modification and purification of peptides on the same resin, reducing the need for extra steps and costly building blocks, and minimizing side reactions, thereby improving the accuracy of peptide analysis and handling.
Implementation Method 1
in a releasing step, adding a reducing agent yielding a reduced intermediate RI and subsequent decomposition of the reduced intermediate RI yielding the modified peptide mP
Data Source
AI summary
The present invention relates to a method for modifying and purifying peptides comprising an immobilizing step, a modification step and a releasing step. In the immobilizing step, a crude linker-tagged peptide L-P is coupled to a solid support yielding an immobilized linker-tagged peptide S-L-P. Subsequently, the immobilized linker-tagged peptide S-L-P is modified with one or more organic molecules yielding an immobilized linker-tagged peptide S-L-mP. Finally, the modified peptide is released via a reduced intermediate RI. The linker molecule is a compound of formula 1, X—Tb—Va-U—Y—Z (1), which can be coupled to a purification resin via the moiety X and to a peptide via the moiety Y under the release of the leaving group Z. T is an optional spacer moiety and V is an optional electron withdrawing moiety. U is an aryl or 5- or 6-membered heteroaryl moiety bound to at least one electron withdrawing moiety V, W or E. The linker is stable under acidic conditions and releases the peptide upon addition of a reducing agent.