De-immunized Factor VIII Protein via Point Mutations
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Solution Overview
Problem
Current recombinant Factor VIII (FVIII) proteins used for treating Hemophilia A often induce an immune response, leading to the formation of inhibitory antibodies, which complicates treatment and can result in reduced efficacy.
Innovation Solution
A recombinant FVIII protein with specific point mutations at defined positions is developed, which reduces the protein's immunogenicity by eliminating T cell epitopes relevant to a significant proportion of humans, while maintaining at least 50% coagulant activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If recombinant FVIII protein is used for treating Hemophilia A, then coagulant activity is provided, but immune response is induced leading to inhibitory antibody formation
Solution Approach 1:
The patent applies local quality by making specific point mutations at defined positions within the FVIII protein sequence. These localized amino acid substitutions target specific T cell epitopes (such as positions in the A2, A3, C1, and C2 domains) without altering the overall protein structure or function. This allows the protein to maintain coagulant activity while reducing immunogenicity at specific locations.
Solution Approach 2:
The patent employs parameter changes by substituting specific amino acids at defined positions in the FVIII sequence. These parameter changes (amino acid substitutions) modify the protein's immunogenic properties while preserving its coagulant function. The mutations are designed to eliminate T cell epitopes that are relevant to a significant proportion of humans, thereby reducing the likelihood of immune response and inhibitory antibody formation.
2Object-generated harmful factors
If point mutations are introduced to eliminate T cell epitopes, then immunogenicity is reduced, but coagulant activity may be compromised
Solution Approach 1:
The patent applies local quality by making specific point mutations at defined positions within the FVIII protein sequence. These localized amino acid substitutions target specific T cell epitopes (such as positions in the A2, A3, C1, and C2 domains) without altering the overall protein structure or function. This allows the protein to maintain coagulant activity while reducing immunogenicity at specific locations.
Solution Approach 2:
The patent employs parameter changes by substituting specific amino acids at defined positions in the FVIII sequence. These parameter changes (amino acid substitutions) modify the protein's immunogenic properties while preserving its coagulant function. The mutations are designed to eliminate T cell epitopes that are relevant to a significant proportion of humans, thereby reducing the likelihood of immune response and inhibitory antibody formation.
3Object-generated harmful factors
If multiple amino acid substitutions are made at defined positions, then T cell epitopes are eliminated, but protein structure complexity increases
Solution Approach 1:
The patent applies local quality by making specific point mutations at defined positions within the FVIII protein sequence. These localized amino acid substitutions target specific T cell epitopes (such as positions in the A2, A3, C1, and C2 domains) without altering the overall protein structure or function. This allows the protein to maintain coagulant activity while reducing immunogenicity at specific locations.
Solution Approach 2:
The patent applies segmentation by dividing the FVIII protein into functional domains (A1, A2, A3, C1, C2) and selectively mutating specific regions within these domains that contain T cell epitopes. This segmented approach allows targeted modification of immunogenic regions while preserving the integrity and function of other critical domains.
Data Source
AI summary
The present invention relates to the field of therapeutic proteins, in particular, to recombinant coagulation factors. It provides a recombinant Factor VIII (FVIII) protein comprising specific point mutations at defined positions, which serve to reduce the immunogenicity of said FVIII protein, wherein the Factor VIII protein substantially retains its coagulant activity. It further provides nucleic acids encoding said de-immunized protein, cell lines and methods of recombinant preparation as well as pharmaceutical compositions comprising the recombinant FVIII of the invention, which are advantageous for use in treatment of patients with Hemophilia A, particularly those who have not yet been treated with a FVIII product. Additionally, it can be a safe alternative for previously treated patients and even for patients who have developed an immune-response to FVIII, e.g., for immune-tolerance-induction therapy (ITI/ITT) or rescue ITI. The invention also provides an assay for determining immunogenicity of a protein.