Freezing of Aqueous Solutions and Chemical Stability of Amorphous Pharmaceuticals: Water Clusters Hypothesis.

Molecular mobility has been traditionally invoked to explain physical and chemical stability of diverse pharmaceutical systems. Although the molecular mobility concept has been credited with creating a scientific basis for stabilization of amorphous pharmaceuticals and biopharmaceuticals, it has become increasingly clear that this approach represents only a partial description of the underlying fundamental principles. An additional mechanism is proposed herein to address 2 key questions: (1) the existence of unfrozen water (i.e., partial or complete freezing inhibition) in aqueous solutions at subzero temperatures and (2) the role of water in the chemical stability of amorphous pharmaceuticals. These apparently distant phenomena are linked via the concept of water clusters. In particular, freezing inhibition is associated with the confinement of water clusters in a solidified matrix of an amorphous solute, with nanoscaled water clusters being observed in aqueous glasses using wide-angle neutron scattering. The chemical instability is suggested to be directly related to the catalysis of proton transfer by water clusters, considering that proton transfer is the key elementary reaction in many chemical processes, including such common reactions as hydrolysis and deamidation.EPSRC EP/N022769/

Interfacial Stress in the Development of Biologics: Fundamental Understanding, Current Practice, and Future Perspective

Biologic products encounter various types of interfacial stress during development, manufacturing, and clinical administration. When proteins come in contact with vapor–liquid, solid–liquid, and liquid–liquid surfaces, these interfaces can significantly impact the protein drug product quality attributes, including formation of visible particles, subvisible particles, or soluble aggregates, or changes in target protein concentration due to adsorption of the molecule to various interfaces. Protein aggregation at interfaces is often accompanied by changes in conformation, as proteins modify their higher order structure in response to interfacial stresses such as hydrophobicity, charge, and mechanical stress. Formation of aggregates may elicit immunogenicity concerns; therefore, it is important to minimize opportunities for aggregation by performing a systematic evaluation of interfacial stress throughout the product development cycle and to develop appropriate mitigation strategies. The purpose of this white paper is to provide an understanding of protein interfacial stability, explore methods to understand interfacial behavior of proteins, then describe current industry approaches to address interfacial stability concerns. Specifically, we will discuss interfacial stresses to which proteins are exposed from drug substance manufacture through clinical administration, as well as the analytical techniques used to evaluate the resulting impact on the stability of the protein. A high-level mechanistic understanding of the relationship between interfacial stress and aggregation will be introduced, as well as some novel techniques for measuring and better understanding the interfacial behavior of proteins. Finally, some best practices in the evaluation and minimization of interfacial stress will be recommended

“pH Swing” in Frozen SolutionsConsequence of Sequential Crystallization of Buffer Components

Succinate buffer solutions of different initial pH values and concentrations were cooled. The solution pH and the phases crystallizing from solution were monitored as a function of temperature. In a solution buffered to pH 4.0 (200 mM), the freeze-concentrate pH initially increased to 8.0 and then decreased to 2.2. On the basis of X-ray diffractometry (synchrotron source), the “pH swing” was attributed to the sequential crystallization of succinic acid, monosodium succinate, and disodium succinate. A similar swing, but in the opposite direction, was seen when a solution with an initial pH of 6.0 was cooled. In this case, crystallization of the basic buffer component occurred first. The direction and magnitude of the pH shift depended on both the initial pH and the buffer concentration. In light of the pH-sensitive nature of a significant fraction of pharmaceuticals (especially proteins), extreme care is needed, both in the buffer selection and in its concentration

Terahertz Dynamics in the Glycerol-Water System

The model glass-former glycerol and its aqueous mixtures were investigated with terahertz-time domain spectroscopy (THz-TDS) in the frequency range of 0.3--3.0\,THz at temperatures from 80--305\,K. It was shown that the infrared absorption coefficient measured with THz-TDS can be theoretically related to the reduced Raman intensity ($\propto \alpha/\omega^2$) and the reduced density of states ($\propto \alpha/\omega^3$) and the agreement with experimental results confirms this. The data were further used to investigate the behaviour of model glasses in the harmonic (below the glass transition temperature $T_{\text{g}}$), anharmonic (above $T_{\text{g}}$), and liquid regime. The onset temperature of the molecular mobility as measured by the infrared active dipoles, $T_{\text{g}}$, was found to correlate with the onset of anharmonic effects, leading to an apparent shift of the boson peak and obscuring it at elevated temperatures. The influence of clustered and unclustered water on the dynamics, the boson peak, and the vibrational dynamics was also investigated. A change in structural dynamics was observed at a water concentration of approximately 5\,wt.\%, corresponding to a transition from isolated water molecules distributed homogeneously throughout the sample to the presence of small water clusters and an increased number of water-water hydrogen bonds which lower the barriers on the potential energy surface.Comment: 13 pages, 8 figure