Antibody Formulation Oxidation Prevention

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Solution Overview

Problem

Protein pharmaceuticals face oxidative degradation issues due to exposure to hydrogen peroxide, light, and metal ions, leading to oxidation of susceptible amino acid residues like methionine and tryptophan, which can impact biological activity and stability during storage and handling.

Innovation Solution

The development of liquid formulations containing a compound, such as 5-hydroxy indole, that prevents oxidation of antibodies by being incorporated into the formulation, thereby maintaining the stability and biological activity of the protein.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If L-tryptophan is added to formulations to prevent oxidation of tryptophan residues, then oxidation of tryptophan is reduced, but L-tryptophan possesses strong absorbance in the UV region making it a primary target during photo-oxidation and can generate ROS that lead to methionine oxidation

Engineering Contradiction:
Improveprevention of tryptophan oxidationVSAvoidROS generation and methionine oxidation
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent removes L-tryptophan from the formulation entirely and replaces it with alternative antioxidants such as L-methionine, L-cysteine, or synthetic compounds. This extraction eliminates the harmful photo-oxidation properties of L-tryptophan while maintaining oxidation protection through alternative means.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces intermediary antioxidant compounds that do not possess strong UV absorbance and do not generate ROS. These intermediaries (such as L-methionine, L-cysteine, or synthetic antioxidants) serve as alternative protective agents that prevent oxidation without the harmful side effects of L-tryptophan.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Reliability

If proteins are exposed to light during processing and storage, then photo-oxidation can occur generating ROS, but avoiding light exposure entirely is not always practical during handling and storage

Engineering Contradiction:
Improveprevention of photo-oxidationVSAvoidhandling and storage convenience
Core Design Contradiction:
ReliabilityVSEase of operation

Solution Approach 1:

The patent incorporates antioxidants into the formulation beforehand, creating a protective system that is already in place before light exposure occurs. This preliminary protection allows the protein to withstand normal light exposure during handling and storage without requiring special light-avoidance measures.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent converts the unavoidable light exposure condition into a non-problematic situation by using antioxidants that either do not absorb UV light or do not generate ROS. This transforms the potential harmful photo-oxidation process into a safe condition where light exposure no longer threatens protein stability.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

3Stability of the object's composition

If standard excipients are used in protein formulations, then formulation stability is maintained, but some excipients can induce ROS-induced oxidation of the protein

Engineering Contradiction:
Improveformulation stabilityVSAvoidROS-induced oxidation
Core Design Contradiction:
Stability of the object's compositionVSObject-generated harmful factors

Solution Approach 1:

The patent uses antioxidants that act as sacrificial protective agents, consuming the oxidation stress themselves to protect the protein. These antioxidants (such as L-methionine, L-cysteine, or synthetic compounds) serve as disposable protective elements that can be replenished or replaced in the formulation.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent changes the chemical parameters of the antioxidant components by selecting compounds with specific properties: they should not possess strong UV absorbance and should not generate ROS. This parameter selection ensures that the antioxidants provide protection without introducing harmful side effects.

Inventive Principle:
Principle #35Parameter changes

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 use of these formulations effectively reduces protein oxidation, ensuring the stability and biological activity of antibodies during storage and handling, addressing the challenges posed by standard excipients like L-tryptophan which can induce ROS-induced oxidation.

Implementation Method 1

Oxidative degradation of amino acid residues is a commonly observed phenomenon in protein pharmaceuticals

Methodology Applied
Scientific EffectOxidation: Oxidation

Implementation Method 2

Light exposure can result in protein oxidation through the formation of reactive oxygen species (ROS) including singlet oxygen, hydrogen peroxide and superoxide

Methodology Applied
Scientific EffectROS generation:

Data Source

PatentEP2968467B1Formulations with reduced oxidation
Publication Date: 2020.06.03 F HOFFMANN LA ROCHE & CO AG
  • EP2968467B1 patent drawingFigure 1A~1B
  • EP2968467B1 patent drawingFigure 2
  • EP2968467B1 patent drawingFigure 3A~3B

AI summary

The invention provides formulations comprising a protein in combination with a compound that prevents oxidation of the protein. The invention also provides methods for making such formulations and methods of using such formulations. The invention further provides methods of screening for compounds that prevent oxidation of a protein in a protein composition and methods of preventing oxidation of a protein in a formulation.