Substituted IL-15 Polypeptides Reducing Deamidation
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Solution Overview
Problem
The ex-vivo manufacture of Interleukin-15 (IL-15) is hindered by deamidation issues, leading to degradation by-products that complicate purification, storage, and clinical testing, necessitating improved IL-15 products with reduced or eliminated deamidation.
Innovation Solution
Substituted IL-15 amino acid sequences and corresponding gene sequences are developed to prevent deamidation, specifically altering residues at positions 71, 72, 77, and 78 to reduce or eliminate deamidation, facilitating refolding, purification, storage, and characterization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If IL-15 is manufactured ex-vivo, then IL-15 product is produced, but deamidation occurs leading to degradation by-products
Solution Approach 1:
The patent applies parameter changes by substituting specific amino acid residues (Asn at positions 7, 10, 12, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100) in the IL-15 polypeptide sequence. These substitutions change the chemical parameters of the protein to reduce susceptibility to deamidation, thereby improving stability while maintaining production capability
Solution Approach 2:
The patent converts the harmful effect of deamidation into a benefit by strategically placing substitutions that actually enhance certain properties. The substitutions at specific positions create a more stable protein structure that resists degradation, turning the vulnerability to deamidation into an opportunity for creating a more robust therapeutic agent
2Manufacturing precision
If deamidation is reduced through substitutions, then purity is improved, but manufacturing complexity increases
Solution Approach 1:
The patent applies local quality by making targeted substitutions at specific positions within the IL-15 sequence rather than throughout the entire molecule. Each substitution is placed strategically at positions most prone to deamidation, creating localized improvements in stability without requiring comprehensive modification of the entire protein structure
Solution Approach 2:
The patent segments the IL-15 molecule into specific regions of interest (positions 7, 10, 12, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100) and applies substitutions selectively to these segments. This segmented approach allows for systematic improvement of purity while managing manufacturing complexity through modular design
Data Source
AI summary
The invention provides IL-15 amino acid sequences with amino acid substitutions that reduce or eliminate deamidation of IL-15 and degradation by-products. The invention also provides DNA sequences that encode the substituted amino acid sequences, a pharmaceutical composition comprising the substituted IL-15 amino acid sequence and a pharmaceutically acceptable carrier, and a method of treating a condition in a mammalian host comprising administering to the host the substituted IL-15 amino acid sequence or the pharmaceutical composition including the substituted IL-15 amino acid sequence in an amount effective to treat the condition in the host.