Acylated Single-Chain Insulin Analogues for Stability
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Solution Overview
Problem
Current insulin formulations face challenges such as thermal fibrillation, chemical degradation, and increased mitogenicity, which limit their stability and efficacy, especially at room temperature, and can lead to unpredictable glucose fluctuations and safety concerns in diabetes management.
Innovation Solution
Development of acylated single-chain insulin analogues with foreshortened connecting domains and specific amino acid modifications, such as fatty acid or dicarboxylic acid tethering, to enhance thermodynamic stability, reduce cross-binding to IGF-1R, and prolong pharmacokinetic and pharmacodynamic profiles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional insulin formulations are used, then insulin activity is maintained, but thermal fibrillation and chemical degradation occur at room temperature
Solution Approach 1:
The patent modifies the molecular structure of insulin by changing amino acid sequences (e.g., replacing B24-B27 segment with alternative sequences), adding fatty acid side chains at specific positions (B3, B28, B29, A14), and creating single-chain variants with foreshortened connecting domains. These parameter changes in protein structure and composition confer resistance to thermal fibrillation and chemical degradation while maintaining biological activity at room temperature
Solution Approach 2:
The patent creates composite molecular structures by combining insulin core with fatty acid side chains (e.g., palmitic acid, stearic acid, arachidic acid) attached at specific amino acid positions. This composite structure integrates the stabilizing effect of hydrophobic fatty acid chains with the biological activity of insulin, providing both thermal stability and functional activity
2Reliability
If single-chain insulin analogues with foreshortened connecting domains are designed, then resistance to thermal degradation is enhanced, but mitogenicity and cross-binding to IGF-1R increase
Solution Approach 1:
The patent introduces fatty acid side chains at specific localized positions (B3, B28, B29, or A14) rather than uniformly throughout the molecule. This local modification provides thermal stability at specific sites while the overall sequence design (foreshortened connecting domain, specific amino acid substitutions) controls mitogenicity and IGF-1R binding properties
Solution Approach 2:
The patent systematically varies amino acid sequences in the connecting domain (reducing from 36 to 5-11 residues) and introduces fatty acid modifications at specific positions to achieve the optimal balance between thermal stability and reduced mitogenicity. Specific substitutions at positions B10, B24, B26, and other locations are used to control receptor binding characteristics
3Duration of action of moving object
If acylation is introduced to prolong pharmacokinetic profile, then duration of action is extended, but manufacturing complexity increases
Solution Approach 1:
The fatty acid side chains are incorporated into the insulin molecule during the peptide synthesis process rather than as a separate post-translational modification step. This preliminary action integrates the acylation into the manufacturing process itself, extending duration of action while avoiding complex downstream modification steps
Data Source
AI summary
A single-chain insulin analogue comprises the insulin B-chain polypeptide sequence, the insulin A-chain polypeptide sequence, and a connecting polypeptide sequence of 5-11 amino acids linking the C-terminal amino acid of the B-chain polypeptide to the N-terminal amino acid of the A-chain polypeptide. The analogue comprises an acetylated Lys at a location selected from the group consisting of any of the amino acids in the connecting polypeptide, B0-B3, B28-B29 or A14, relative to wild type insulin, or comprises an acetylated amino acid at the N-terminal amino acid of the single-chain insulin analogue. The single-chain insulin analogue may be acylated with a C6-C21 fatty acid, which may be attached to the e-amino group of a unique Lysine residue or the a-amino group of the N-terminal amino acid of the single-chain insulin analogue. The insulin analogue may be used to lower the blood sugar of a patient in need thereof.