TNFR:Fc Liquid Formulation Stabilization via Excipient Matrix
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Solution Overview
Problem
Proteins like TNFR:Fc face challenges in maintaining stability during long-term storage due to denaturation, aggregation, and fragmentation, especially when exposed to temperature and physical stress, which affects their biological activity and requires improved formulations for enhanced stability and convenience.
Innovation Solution
A novel liquid formulation comprising TNFR:Fc with aspartic acid as an aggregation inhibitor, phosphate buffer, polysorbate 20 as a surfactant, sucrose as a stabilizer, and EDTA as a fragmentation inhibitor, along with sodium chloride for tonicity, to create a stable aqueous solution that maintains activity at refrigerated and room temperatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If proteins are stored in aqueous formulations at refrigerated temperatures, then their storage convenience is improved, but they still undergo denaturation and aggregation over time
Solution Approach 1:
The patent introduces multiple excipients as intermediary substances that mediate between the protein and the aqueous environment. Polysorbate 20 acts as a surfactant intermediary that prevents protein aggregation by adsorbing at protein-protein interfaces. Phosphate buffer serves as a chemical intermediary that maintains pH stability. These intermediary substances protect the protein from denaturation while allowing storage in convenient aqueous form at refrigerated temperatures.
Solution Approach 2:
The patent optimizes multiple formulation parameters simultaneously: pH is adjusted to 6.0 using phosphate buffer, surfactant concentration is set at 0.05% polysorbate 20, and the formulation maintains specific ionic strength. By carefully controlling these parameters, the protein achieves enhanced stability in liquid form at 4°C, resolving the contradiction between storage convenience and protein stability.
2Reliability
If lyophilization is used to stabilize proteins for long-term storage, then protein stability is improved, but the formulation requires reconstitution before use
Solution Approach 1:
The patent extracts the stability-enhancing properties of lyophilization (use of stabilizing excipients like sugars and surfactants) while eliminating the need for freeze-drying and reconstitution. By incorporating polysorbate 20 and phosphate buffer in optimized concentrations, the formulation achieves long-term stability in liquid form, taking out the beneficial stabilization mechanism while removing the inconvenient reconstitution step.
Solution Approach 2:
The formulation includes pre-optimized concentrations of stabilizing excipients (polysorbate 20 at 0.05%, phosphate buffer at pH 6.0) that are prepared in advance during manufacturing. This preliminary incorporation of stabilizing agents ensures protein stability throughout storage without requiring any action from the user, eliminating the reconstitution step while maintaining reliability.
3Quantity of substance
If protein concentration is increased to reduce formulation volume, then storage efficiency is improved, but aggregation and denaturation occur more rapidly
Solution Approach 1:
The patent uses polysorbate 20 as a surfactant intermediary that becomes particularly important at higher protein concentrations. The surfactant adsorbs at protein-protein interfaces, preventing aggregation even when proteins are in close proximity at high concentrations. This allows the formulation to achieve both high storage efficiency and maintained protein stability.
Solution Approach 2:
The patent optimizes the pH parameter to 6.0 using phosphate buffer, which is near the isoelectric point of many proteins but prevents aggregation through electrostatic repulsion. This parameter optimization allows higher protein concentrations to be maintained without rapid aggregation, improving storage efficiency while preserving protein stability.
4Reliability
If excipients are added to improve protein stability, then protein stability is improved, but formulation complexity increases
Solution Approach 1:
The patent employs excipients that perform multiple functions: phosphate buffer maintains pH stability and provides ionic strength; polysorbate 20 prevents aggregation and acts as a surfactant; sodium chloride adjusts tonicity and influences protein solubility. By selecting excipients with multi-functional properties, the formulation achieves enhanced protein stability without unnecessarily increasing complexity.
Solution Approach 2:
The patent creates a composite aqueous formulation where the protein is combined with a specific matrix of excipients (polysorbate 20, phosphate buffer, sodium chloride) in optimized proportions. This composite approach allows the system to achieve enhanced stability through synergistic interactions among components while maintaining a relatively simple overall formulation structure that can be manufactured using standard processes.
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 formulation achieves long-term stability and bioactivity of TNFR:Fc, reducing aggregation and fragmentation, and providing enhanced shelf life even at elevated temperatures, thus addressing the challenges of protein stability and storage convenience.
Implementation Method 1
an aggregation inhibitor, a buffer, a non-ionic surfactant, a stabilizer
Implementation Method 2
a buffer maintaining the pH in the range from about 4.5 to about 6.0
Implementation Method 3
a surfactant and a polyol
Implementation Method 4
a stabilizer, a tonicity modifier, and optionally a preservative
Implementation Method 5
EDTA as a fragmentation inhibitor
Data Source
Figure 1a~3b
Figure 4~7b
Figure 8a~8c
AI summary
The present invention relates to a liquid formulation of a polypeptide containing an Fc domain of an immunoglobulin which is stabilized to maintain the activity of a polypeptide containing an Fc domain for a prolonged period of time.