VEGF-A Polypeptides with Rearranged Disulfide Bonds

Solution Overview

Problem

Current strategies for targeting VEGF-A in cancer and inflammatory diseases have limited efficacy due to marginal immune responses and adverse effects, with xenogeneic variants inducing neutralizing antibodies but not providing significant anti-tumoral or anti-metastatic effects, and homologous variants resulting in minimal immune activation.

Innovation Solution

Development of polypeptides comprising functional mutants of human VEGF-A isoforms with non-natural disulfide bridge arrangements, specifically where the 2nd and 4th cysteines form intramolecular bonds and the 7th and 8th cysteines form intermolecular bonds, enhancing immunogenicity and antitumoral effects while avoiding signal transduction associated with VEGFR2 binding.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If xenogeneic variants of VEGF-A are used, then neutralizing antibody titers are induced, but anti-tumoral and anti-metastatic effects are marginal

Engineering Contradiction:

Improveneutralizing antibody responseVSAvoidlack of anti-tumoral effect

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent applies local quality by creating specific mutations at defined locations in the VEGF-A protein sequence (positions 82, 84, 86) to alter binding properties. The mutations are strategically placed to disrupt VEGFR2 binding while preserving other functional aspects, thereby achieving localized functional modification rather than global changes.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the biochemical parameters of VEGF-A by introducing specific amino acid mutations (Arg82, Lys84, His86→Ala/Glu) that modify the protein's binding affinity and immunogenicity. These parameter changes transform the protein from a natural ligand into a modified variant with enhanced immunogenic properties and reduced binding to VEGFR2.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If homologous variants of VEGF-A are used, then immune response is induced, but anti-tumoral effects are marginal

Engineering Contradiction:

Improveimmune responseVSAvoidlack of anti-tumoral effect

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent applies local quality by creating specific mutations at defined locations in the VEGF-A protein sequence (positions 82, 84, 86) to alter binding properties. The mutations are strategically placed to disrupt VEGFR2 binding while preserving other functional aspects, thereby achieving localized functional modification rather than global changes.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the biochemical parameters of VEGF-A by introducing specific amino acid mutations (Arg82, Lys84, His86→Ala/Glu) that modify the protein's binding affinity and immunogenicity. These parameter changes transform the protein from a natural ligand into a modified variant with enhanced immunogenic properties and reduced binding to VEGFR2.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If VEGF-A binds to VEGFR2, then signal transduction occurs, but anti-angiogenic effects are reduced

Engineering Contradiction:

Improvesignal transductionVSAvoidreduced anti-angiogenic effect

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent extracts the binding function from the VEGF-A protein by introducing mutations that specifically disrupt the interaction with VEGFR2. The mutations at positions 82, 84, and 86 effectively remove or reduce the protein's ability to bind to its receptor, thereby extracting the signaling function while preserving the protein's structural integrity and immunogenicity.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent inverts the normal function of VEGF-A by creating a variant that cannot bind to VEGFR2. Instead of promoting signal transduction and angiogenesis, the mutated protein serves as an immunogen that induces antibodies against the natural VEGF-A/VEGFR2 interaction, effectively inverting the biological outcome from pro-angiogenic to anti-angiogenic through immune mediation.

Inventive Principle:

Principle #13The other way round (Inversion)

Data Source

PatentUS20230046501A1Polypeptides comprising mutated forms of human VEGF-a with rearrangements of disulfide bonds and compositions containing same

Publication Date: 2023.02.16 CENT DE ING GENETICA & BIOTECNOLOGIA

AI summary

Polypeptides comprising functional mutants of an isoform of the human vascular endothelial growth factor A (VEGF-A) folded in a non-natural re-arrangement, where the second and fourth cysteine of the mutant's polypeptide chain is only forming intramolecular bridges, while the seventh and eight are only part of intermolecular bonds. The invention further comprises antigenic preparations containing at least one of these polypeptides, and the pharmaceutical compositions comprising such antigenic preparations and vaccine adjuvants. The antigenic preparations according to the invention are used in the manufacturing of a drug, for the treatment of diseases related to the increment of angiogenesis, inflammation, and immunosuppression, as well as for the restoration of the immune system.