PH20 Polypeptide Variants for Preservative-Resistant Stability
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Solution Overview
Problem
Existing hyaluronidases, such as bovine and ovine forms, are immunogenic for human treatment and lack sufficient stability under denaturing conditions, limiting their therapeutic efficacy.
Innovation Solution
Modified PH20 polypeptides with specific amino acid replacements or modifications that enhance stability and activity, including resistance to protein denaturation conditions like elevated temperature and preservatives, while maintaining at least 40% of the unmodified PH20 polypeptide's activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing hyaluronidases (bovine and ovine forms) are used for human treatment, then they can provide hyaluronidase activity, but they are immunogenic and lack sufficient stability under denaturing conditions
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of PH20 hyaluronidase through specific substitutions (e.g., replacing bovine/ovine sequences with human-compatible sequences). This changes the chemical parameters of the enzyme to reduce immunogenicity while maintaining or enhancing stability under denaturing conditions including phenolic preservatives and elevated temperatures.
Solution Approach 2:
The patent creates composite enzymatic structures by combining stabilized PH20 polypeptide variants with phenolic preservatives in formulations. The polypeptide variants are engineered to resist denaturation by these preservatives, creating a stable composite therapeutic formulation that maintains activity in the presence of stabilizing agents.
2Reliability
If PH20 polypeptide variants with amino acid replacements are created to enhance stability, then stability under denaturing conditions improves, but complexity of polypeptide structure increases
Solution Approach 1:
The patent applies local quality by making targeted amino acid replacements at specific positions within the PH20 polypeptide chain rather than uniformly modifying the entire structure. Specific residues are substituted to confer resistance to phenolic preservatives and thermal denaturation, while leaving the rest of the protein structure intact to maintain native function with minimal complexity increase.
Solution Approach 2:
The patent changes specific physical-chemical parameters of the polypeptide through amino acid substitutions, such as altering hydrophobicity, charge distribution, or steric properties at key positions. These parameter changes enhance stability under denaturing conditions while maintaining relatively simple polypeptide structures that retain hyaluronidase activity.
3Reliability
If phenolic preservatives are added to formulations, then stability of polypeptide is improved, but harmful effects from preservatives increase
Solution Approach 1:
The patent converts the potentially harmful effect of phenolic preservatives into a beneficial stabilizing force. PH20 polypeptide variants are engineered to specifically resist denaturation by phenolic preservatives, transforming these preservatives from harmful denaturing agents into beneficial stability-enhancing components of the formulation.
Solution Approach 2:
The patent modifies the polypeptide's resistance parameters to phenolic preservatives through amino acid replacements, changing the interaction parameters between the enzyme and preservative. This allows the formulation to tolerate higher preservative concentrations for stability without increasing harmful effects, as the modified polypeptide is specifically engineered to withstand these conditions.
Data Source
AI summary
Modified PH20 hyaluronidase polypeptides, including modified polypeptides that exhibit increased stability and/or increased activity, are provided. Also provided are compositions and formulations and uses thereof.