GRF Formulation Stabilization via Beta-Cyclodextrin Inclusion Complexes
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Solution Overview
Problem
There is a need for improved pharmaceutical formulations of GRF molecules that maintain physical and chemical integrity during manufacturing, storage, and use, as existing formulations face issues with proteolytic stability, aggregation, denaturation, oxidation, and deamidation, leading to loss of bioactivity over time.
Innovation Solution
A liquid pharmaceutical formulation comprising [trans-3-hexenoyl]hGRF(1-44) amide with a modified β-cyclodextrin at concentrations of 5-10% (w/v) and a pH of 5.5-6.5, which is not conjugated to the GRF molecule, along with additional components like bulking agents and anti-microbial agents, to enhance stability and bioactivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If GRF molecules are stored in conventional formulations, then they can be used for treatment, but they undergo deamidation, aggregation, and denaturation leading to loss of bioactivity over time
Solution Approach 1:
The patent introduces modified beta-cyclodextrin as an intermediary stabilizing agent that forms inclusion complexes with the GRF molecule. The cyclodextrin's hydrophobic cavity encapsulates hydrophobic regions of the peptide, protecting it from deamidation and aggregation. This mediator approach resolves the contradiction by enabling long-term storage without compromising molecular stability.
Solution Approach 2:
The patent optimizes multiple formulation parameters including pH (5.0-6.0), ionic strength, and temperature to minimize deamidation and aggregation. By carefully controlling these parameters and using modified beta-cyclodextrin derivatives with specific substitution patterns, the formulation maintains GRF bioactivity throughout the storage period, resolving the stability-duration contradiction.
2Reliability
If GRF formulations are stored at refrigerated temperatures, then stability is improved, but accessibility and ease of use are reduced
Solution Approach 1:
The patent develops lyophilized (freeze-dried) GRF formulations that can be stored at room temperature in stable, dry form. The product is supplied as a powder or capsule that requires no refrigeration, making it easily accessible and transportable. Stability is achieved through the combination of lyophilization and modified beta-cyclodextrin, which protects the GRF molecule during storage and handling.
Solution Approach 2:
The patent utilizes lyophilization (freeze-drying) to transition the GRF formulation from liquid to solid state. This phase transition removes water that would otherwise promote deamidation and microbial growth, enabling room temperature storage. The modified beta-cyclodextrin maintains molecular stability in the dry state, resolving the contradiction between accessibility and integrity.
3Productivity
If GRF molecules are concentrated to improve dosing efficiency, then injection volume is reduced, but aggregation and denaturation risk increases
Solution Approach 1:
The modified beta-cyclodextrin acts as a protective intermediary that forms soluble inclusion complexes with GRF even at high concentrations. The cyclodextrin's hydrophobic cavity shields the peptide from self-aggregation, allowing the formulation to achieve high dosing efficiency without compromising physical stability. This enables concentrated formulations that maintain monomeric GRF structure.
4Ease of manufacture
If pH is adjusted to optimize solubility, then formulation ease of manufacture is improved, but chemical stability through deamidation is compromised
Solution Approach 1:
The patent carefully optimizes pH to the narrow range of 5.0-6.0, which balances solubility and chemical stability. At this pH range, the GRF molecule remains sufficiently soluble for formulation while minimizing deamidation of asparagine residues. The modified beta-cyclodextrin further stabilizes the molecule chemically, allowing the formulation to achieve both manufacturability and stability without requiring extreme pH adjustments.
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 provides high stability of the GRF molecule, with at least 70% of the GRF molecule remaining non-deamidated after 2 years at refrigerated temperatures and 3 years at room temperature, maintaining bioactivity and efficacy for extended storage periods.
Implementation Method 1
GRF analogs containing a hydrophobic tail as defined in the present application are modified versions or analogs of human GRF that have been shown to have higher proteolytic stability in biological milieu
Implementation Method 2
a liquid pharmaceutical formulation comprising: [trans-3-hexenoyl]hGRF(1-44) amide... and a modified β-cyclodextrin
Implementation Method 3
wherein said formulation has a pH of 5.5 to 6.5... at least 70% of the GRF molecule remaining non-deamidated after 2 years at refrigerated temperatures and 3 years at room temperature
Data Source
Figure 1~3
Figure 4
Figure 5~6
AI summary
Stabilized solid and liquid pharmaceutical formulations comprising a GRF molecule as active ingredient, such as GRF analogs including those comprising an N-terminal-attached hydrophobic moiety, such as [trans-3-hexenoyl]hGHRH (1-44) amide, are disclosed. The formulation comprises a GRF molecule or a pharmaceutically acceptable salt thereof and a β- cyclodextrin which is not conjugated to the GRF molecule or salt thereof. Also disclosed is the use of the formulation for the treatment of various conditions, methods of preparing the formulation, as well as kits containing it. Methods of stabilizing (e.g., with respect to chemical stability) such GRF molecules, as well as methods of inhibiting their deamidation at Asn8, are also disclosed.