Stapled Polypeptides Stabilizing Alpha-Helical Conformation
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Solution Overview
Problem
Alpha-helical peptides are prone to unraveling and proteolytic degradation, limiting their stability and biological activity as therapeutic agents.
Innovation Solution
The development of stapled and stitched polypeptides using ring-closing metathesis (RCM) to cross-link olefin-containing sidechains, stabilizing the alpha-helical conformation and enhancing resistance to proteolytic cleavage and membrane penetration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If alpha-helical peptides are used as therapeutic agents, then biological activity is achieved, but stability is poor due to unraveling and proteolytic degradation
Solution Approach 1:
The patent applies preliminary action by pre-installing olefin-containing side chains at specific positions (i-1 and i+4) in the amino acid sequence before the actual stapling reaction. This preliminary placement of reactive groups enables the subsequent ring-closing metathesis to efficiently form the stabilizing crosslink without requiring complex in-situ generation of the crosslinking agents, thus resolving the contradiction by preparing the molecular structure in advance for stabilization while maintaining biological activity.
Solution Approach 2:
The patent creates a composite structure by combining the natural peptide backbone with synthetic olefin-containing side chains that form cyclic crosslinks through ring-closing metathesis. This composite approach integrates the biological recognition capabilities of natural peptides with the structural stability of synthetic cyclic constraints, achieving both biological activity and resistance to proteolytic degradation simultaneously.
2Stability of the object's composition
If crosslinking is applied to stabilize alpha-helical conformation, then resistance to proteolytic cleavage increases, but membrane penetration capability may be reduced
Solution Approach 1:
The patent applies local quality by implementing crosslinks at specific local positions within the alpha-helix (at residues i and i+4) rather than uniform crosslinking throughout the entire peptide structure. This localized stapling approach stabilizes the critical helical conformation at key positions while leaving other regions of the peptide flexible and accessible for membrane interaction, thus resolving the contradiction between conformational stability and membrane penetration.
3Stability of the object's composition
If multiple stapling events are performed to provide multiply stapled polypeptide, then structural stability is enhanced, but manufacturing complexity increases
Solution Approach 1:
The patent applies preliminary action by pre-installing multiple olefin-containing side chains at predetermined positions (including i-1, i, i+4 combinations) before the ring-closing metathesis reaction. This preliminary placement strategy enables sequential or simultaneous formation of multiple crosslinks without requiring complex step-by-step synthesis procedures, thus reducing manufacturing complexity while achieving enhanced structural stability through multiple stapling events.
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
Stabilization of the alpha-helical conformation and improved biological activity of the peptides, leading to enhanced therapeutic potential for proliferative, neurological, and other disorders.
Implementation Method 1
two olefin-containing sidechains present in a polypeptide are covalently joined ('stapled') using a ring-closing metathesis (RCM) reaction to form a crosslink
Data Source
AI summary
The present invention provides stapled polypeptides of the Formulae (I) and (VI):and salts thereof; wherein the groups ; R1a, R1b, R1c, R2a, R3a, R2b, R3b, R4a, R4b, RA, RZ, L1a, L1b, L2, L3, XAA, v, w, p, m, s, n, t, and q are as defined herein. The present invention further provides methods of preparing the inventive stapled polypeptides from unstapled polypeptide precursors. The present invention further provides pharmaceutical compositions comprising a stapled polypeptide of Formula (I) or (VI), and methods of using the stapled peptides. The present invention also provides modifications of the staples post ring closing metathesis.