Buffered Bevacizumab Formulation Stabilization

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Solution Overview

Problem

Bevacizumab antibodies, such as those marketed under the brand Avastin, tend to aggregate during storage, leading to reduced potency due to covalent aggregate formation in the antigen-binding domain, which diminishes their effectiveness in binding to vascular endothelial growth factor (VEGF).

Innovation Solution

A buffered formulation of bevacizumab is developed, comprising specific concentrations of citrate phosphate, trehalose or sucrose, and polysorbate 20, with a pH range of 5.6 to 6.1, which stabilizes the antibody, reducing aggregation and maintaining its VEGF binding activity over extended storage periods.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If bevacizumab is stored under conventional conditions, then the antibody remains stable initially, but aggregates form over time reducing potency

Engineering Contradiction:
Improvestorage stabilityVSAvoidaggregate formation
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent applies parameter changes by optimizing the pH to a specific range of 5.6-6.1 and adjusting buffer composition (citrate phosphate, sodium phosphate) along with stabilizer concentrations (trehalose 50-150 mg/mL, polysorbate 20 0.02-0.1%) to minimize aggregate formation and maintain antibody stability during long-term storage

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses buffer components and stabilizers as intermediary substances that mediate between the antibody and storage conditions. Specifically, citrate phosphate and sodium phosphate buffers along with trehalose and polysorbate 20 act as protective intermediaries that prevent direct harmful interactions leading to aggregation

Inventive Principle:
Principle #24Intermediary (Mediator)

2Stability of the object's composition

If the formulation uses higher stabilizer concentrations to prevent aggregation, then colloidal stability improves, but formulation complexity increases

Engineering Contradiction:
Improvecolloidal stabilityVSAvoidformulation complexity
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent optimizes stabilizer concentrations within specific ranges (trehalose 50-150 mg/mL, polysorbate 20 0.02-0.1%) rather than using excessive amounts, achieving colloidal stability while maintaining formulation simplicity through parameter optimization

Inventive Principle:
Principle #35Parameter changes

3Quantity of substance

If the antibody concentration is increased to improve therapeutic efficacy, then potency per volume increases, but aggregation tendency increases

Engineering Contradiction:
Improveantibody concentrationVSAvoidaggregation
Core Design Contradiction:
Quantity of substanceVSStability of the object's composition

Solution Approach 1:

The patent enables higher antibody concentrations to be maintained in solution by changing the pH parameter to 5.6-6.1 and optimizing buffer and stabilizer concentrations, which suppresses aggregation even at elevated antibody levels, thereby achieving both high potency and stability

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 buffered formulation significantly enhances the thermal and colloidal stability of bevacizumab, ensuring minimal aggregation and maintaining therapeutic efficacy for at least 18 months when stored under refrigerated conditions, thereby improving the long-term storage and administration of the antibody.

Implementation Method 1

The invention features buffered formulations for storage of bevacizumab... the formulation has a pH of 5.6 to 5.8... the buffer may comprise citrate phosphate or sodium acetate

Methodology Applied
Scientific EffectBuffering:

Implementation Method 2

the formulation may also comprise a stabilizer that comprises a sugar such as trehalose or sucrose... enhances the thermal stability and colloidal stability of the antibody

Methodology Applied
Scientific EffectPreferential exclusion:

Implementation Method 3

the formulation may also comprise a mild surfactant such as polysorbate 20... reducing aggregation and maintaining its VEGF binding activity

Methodology Applied
Scientific EffectSurfactant: Surfactant

Data Source

PatentEP3397281B1Buffered formulations of bevacizumab
Publication Date: 2020.09.09 OUTLOOK THERAPEUTICS INC
  • EP3397281B1 patent drawingFigure 1
  • EP3397281B1 patent drawingFigure 2A
  • EP3397281B1 patent drawingFigure 2A(i)

AI summary

The invention provides buffered aqueous formulations of bevacizumab. The formulations comprise a citrate phosphate buffer comprising trehalose or sucrose, and polysorbate 20. The formulations have an acidic pH of from about 5.8 to about 6.0, and enhance the conformational and colloidal stability of the bevacizumab molecule. Levels of potency-reducing reversible aggregates and covalent dimers in the citrate phosphate buffer were low, and this buffer further offered protection against storage stress, but did not modify or alter any key biophysical descriptor of the antibody.