Variant Asparaginase Polypeptides With Reduced Immunogenicity
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Solution Overview
Problem
Existing asparaginase treatments for acute lymphoblastic leukemia and lymphoma face issues of immunogenicity, hypersensitivity, and inefficient production methods, leading to treatment termination and product shortages.
Innovation Solution
Development of variant asparaginase polypeptides with specific amino acid substitutions, particularly at cysteine residues, to reduce immunogenicity and enhance stability, combined with site-specific pegylation and production in host cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If native asparaginase is used for treatment, then therapeutic effect is achieved, but immunogenicity and hypersensitivity occur
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of asparaginase through site-directed mutagenesis. Specifically, cysteine residues at positions 29, 169, and 206 are substituted with serine, threonine, or alanine, respectively. These parameter changes in the protein structure reduce immunogenicity while preserving catalytic activity, thereby resolving the contradiction between therapeutic effect and immunogenicity.
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific locations (cysteine residues) within the asparaginase molecule rather than altering the entire protein. The cysteine residues at positions 29, 169, and 206 are specifically substituted to reduce immunogenicity, while the rest of the protein structure remains unchanged to maintain therapeutic function.
2Reliability
If native asparaginase is used for treatment, then therapeutic effect is achieved, but treatment termination due to hypersensitivity occurs
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of asparaginase through site-directed mutagenesis. Specifically, cysteine residues at positions 29, 169, and 206 are substituted with serine, threonine, or alanine, respectively. These parameter changes in the protein structure reduce immunogenicity while preserving catalytic activity, thereby resolving the contradiction between therapeutic effect and immunogenicity.
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific locations (cysteine residues) within the asparaginase molecule rather than altering the entire protein. The cysteine residues at positions 29, 169, and 206 are specifically substituted to reduce immunogenicity, while the rest of the protein structure remains unchanged to maintain therapeutic function.
3Quantity of substance
If traditional production methods are used, then asparaginase is produced, but production efficiency is low and product shortages occur
Solution Approach 1:
The patent applies self-service by utilizing the host cell's (E. coli) own metabolic pathways and protein expression machinery to produce the variant asparaginase. The mutated gene is introduced into E. coli, which then automatically transcribes and translates the gene to produce the therapeutic protein, eliminating the need for complex external production systems and significantly improving production efficiency.
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
The variant asparaginase polypeptides demonstrate reduced immunogenicity, improved stability, and enhanced pharmacokinetics, addressing hypersensitivity and production inefficiencies, thereby providing effective treatment options for leukemia and lymphoma.
Implementation Method 1
L-asparaginase (also referred to herein as asparaginase) is a bacterial enzyme composed of identical subunits (approximately 35 kDa each subunit) that catalyzes the conversion of L-asparagine to aspartic acid and ammonia
Data Source
Figure 1A~1B
Figure 2A~2B
Figure 3
AI summary
Provided are various embodiments relating to variant asparaginase polypeptides with enhanced stability, pharmacodynamics, and/or reduced immunogenicity. The variant asparaginase polypeptides may be used as therapeutics in mammals, including human, canines, felines, and equines.