Bispecific Antibody Stabilization via Cyclodextrin Mediator

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Solution Overview

Problem

Protein pharmaceuticals face significant challenges due to their marginal stability and susceptibility to degradation, particularly protein aggregation, which affects biological activity, product yield, and can lead to adverse immunogenic reactions, especially during refolding, purification, sterilization, shipping, and storage processes.

Innovation Solution

A stable pharmaceutical composition comprising a bispecific single chain antibody construct, sulfobutylether-β-cyclodextrin sodium salt, and a buffer, optimized with excipients like sucrose and polysorbate 80, to reduce protein aggregation and enhance colloidal stability, thereby preventing loss-of-function and adverse immunogenic reactions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If proteins are manufactured with high purity through commercial scale purification processes, then purity is improved, but proteins remain marginally stable and highly susceptible to degradation

Engineering Contradiction:
ImprovepurityVSAvoidstability
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

Beta-cyclodextrin acts as a stabilizing agent that binds to hydrophobic regions of the antibody construct, preventing aggregation and degradation. The cyclodextrin molecule serves as an intermediary between the protein and the aqueous environment, reducing unwanted interactions and maintaining colloidal stability throughout the shelf life of the product.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Manufacturing precision

If protein aggregation is removed through elaborate purification steps in downstream processing, then aggregation is reduced, but product yield decreases substantially

Engineering Contradiction:
Improveaggregation removalVSAvoidproduct yield
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The formulation includes beta-cyclodextrin from the outset to prevent aggregation during storage and handling, rather than requiring subsequent purification steps to remove aggregates. This preliminary stabilization action maintains product integrity throughout the supply chain, eliminating the need for yield-reducing polishing steps.

Inventive Principle:
Principle #10Preliminary action

3Reliability

If proteins are stabilized by modifying their primary structure through genetic engineering, then stability is improved, but functionality may be impaired and immunogenicity increased

Engineering Contradiction:
ImprovestabilityVSAvoidimmunogenicity
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

Instead of modifying the protein's primary structure, the invention changes the formulation parameters by adding beta-cyclodextrin. This external stabilization approach maintains the natural amino acid sequence and structure of the antibody construct, preserving its biological activity and minimizing immunogenicity while achieving enhanced stability.

Inventive Principle:
Principle #35Parameter changes

4Reliability

If protein aggregation occurs during refolding, purification, sterilization, shipping, and storage processes, then biological activity is impaired, but these processes are necessary for product manufacturing and distribution

Engineering Contradiction:
Improvebiological activityVSAvoidprocess necessity
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The formulation incorporates beta-cyclodextrin as a protective agent that cushions the antibody construct against aggregation during stressful processes such as refolding, purification, sterilization, shipping, and storage. This prior protection allows necessary manufacturing and distribution processes to proceed without compromising product integrity or biological activity.

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

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 composition effectively stabilizes the bispecific single chain antibody construct, reducing protein aggregation and maintaining stability under various stress conditions, including thermal and surface-induced stress, thereby ensuring the biological activity and safety of the therapeutic protein.

Implementation Method 1

a stable pharmaceutical composition comprising a bispecific single chain antibody construct, sulfobutylether-β-cyclodextrin sodium salt, and a buffer, optimized with excipients like sucrose and polysorbate 80, to reduce protein aggregation and enhance colloidal stability

Methodology Applied
Scientific EffectCyclodextrin stabilization: Solvation

Data Source

PatentEP3408295B1Pharmaceutical composition comprising bispecific antibody constructs
Publication Date: 2023.07.26 AMGEN RESEARCH (MUNICH) GMBH
  • EP3408295B1 patent drawingFigure 1
  • EP3408295B1 patent drawingFigure 2A~2B
  • EP3408295B1 patent drawingFigure 2C~3

AI summary

The present invention provides novel and stable pharmaceutical compositions comprising bispecific single chain antibody constructs, cyclodextrins and a buffer.