Freeze-Dry Microscopy of Protein/Sugar Mixtures: Drying Behavior, Interpretation of Collapse Temperatures and a Comparison to Corresponding Glass Transition Data

Meister, Eva; Gieseler, Henning

doi:10.1002/jps.21586

Cited by 99 publications

(68 citation statements)

References 44 publications

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“…19,20) FDM analysis of frozen disaccharide solutions has shown onset of the collapse (T c ) at temperatures several degrees higher than T g ′.…”

Section: )mentioning

confidence: 99%

“…A similar dependence of the T c on the solute concentration has been reported in frozen disaccharide systems. 20,24) A spatially dense solid that forms upon drying of the higher solute concentration solutions would be a possible explanation for the increasing resistance to loss of the cake structure.…”

Section: )mentioning

confidence: 99%

“…1) Recent advances in freeze-drying microscopy (FDM) analysis have enabled direct observation of the collapse phenomena of frozen solutions in a small reduced-pressure cell that mimic the primary drying process. 19,20) FDM analysis of frozen disaccharide solutions has shown onset of the collapse (T c ) at temperatures several degrees higher than T g ′.…”

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confidence: 99%

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Component Crystallization and Physical Collapse during Freeze-Drying of L-Arginine–Citric Acid Mixtures

Yamaki

Ohdate

Nakadai

et al. 2012

CHEMICAL & PHARMACEUTICAL BULLETIN

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Component crystallization and physical collapse during freeze-drying of aqueous solutions containing protein-stabilizing L-arginine and citric acid mixtures were studied. Freeze-drying microscopy (FDM) and thermal analysis of the solute-mixture frozen solutions showed collapse onset at temperatures (T c ) approximately 10°C higher than their T g ′s (glass transition temperatures of the maximally freeze-concentrated solute phase). Experimental freeze-drying of these solutions at a low chamber pressure showed the occurrence of physical collapse at shelf temperatures close to or slightly higher than the T c . Slower ice sublimation at higher chamber pressures induced the physical collapse from lower shelf temperatures. The large effect of chamber pressures on the collapse-inducing shelf temperatures confirmed significance of the sublimation-related heat loss on the sublimation interface temperature during the primary drying. Drying of the single-solute L-arginine solution resulted in cake-structure solids composed of its anhydrous crystal. Thermal and powder X-ray diffraction (PXRD) analysis suggested slow crystal nucleation of L-arginine dihydrate in the frozen solutions. Characterization of the frozen solutions and freeze-dried solids should enable rational formulation design and process control of amino acid-containing lyophilized pharmaceuticals.Key words freeze-drying; crystallization; glass; lyophilization; collapse; thermal analysis Clinical applications of protein pharmaceuticals such as therapeutic antibodies are increasing. However, marginal storage stabilities of many proteins in the aqueous solutions emphasize the importance of developing freeze-dried formulations. Many lyophilized protein formulations contain disaccharides (e.g., sucrose and trehalose) that protect the proteins thermodynamically from structural changes during the freeze-drying process and kinetically from chemical degradation during subsequent storage.1,2) Some basic amino acid (e.g., L-arginine and L-histidine) and multivalent acid (e.g., H 3 PO 4 and citric acid) mixtures are practical alternative stabilizers for protein formulations.3-6) The mixtures protect proteins from conformation-altering dehydration stress during the process. These mixtures form glass-state amorphous solids through networks of hetero-component molecular interactions between the amino and carboxyl groups, and thus improve chemical stability of the dried proteins during storage. 5,7)L-Arginine also increases solubility of various proteins without apparent changes in their conformation. [8][9][10] Freeze-drying is a typical high-energy process, and varying its parameters significantly varies the formulation quality, including the cake appearance, component crystallinity, residual water, and stability of the active pharmaceutical ingredients (APIs).11,12) Failure of a biopharmaceutical lyophilization batch would lead to the loss of the therapeutic protein often more expensive than small molecular APIs. Freezing of aqueous solutions concentrates most solutes to a s...

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“…19,20) FDM analysis of frozen disaccharide solutions has shown onset of the collapse (T c ) at temperatures several degrees higher than T g ′.…”

Section: )mentioning

confidence: 99%

Section: )mentioning

confidence: 99%

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Component Crystallization and Physical Collapse during Freeze-Drying of L-Arginine–Citric Acid Mixtures

Yamaki

Ohdate

Nakadai

et al. 2012

CHEMICAL & PHARMACEUTICAL BULLETIN

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“…In the lyostat2 analysis of both types of formulations drying front progressed at high speed with no product collapse up to -20.0°, material got completely dried below -19.7°. This type of phenomenon resulted due to absence of any water mo bility in the matrix [25] .…”

Section: Resultsmentioning

confidence: 99%

Design of the Lyophilization Process of a Doxorubicin Formulation Based on Thermal Properties

Hajare¹,

More²

2017

pharmaceutical-sciences

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The frontline chemotherapy used for treatment of several forms of cancer is doxorubicin, which is required to be lyophilized due to its limited storage stability. Results from differential thermal analysis/ impedance and freeze drying microscopy determinations, signifies that the freezing step product temperature must be well below -59.8°, while throughout the primary drying stage product temperature at sublimation front cannot exceed -19.7°. The use of a procedure based on the critical parameters allowed freeze drying of doxorubicin laboratory scale batches that would help to reduce process cycle time, energy and costs at commercial production.

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“…Starting the vacuum drying before sufficient crystal growth resulted in a partially collapsed solid due to a higher product temperature than T c of the freeze-concentrated amorphous erythritol region, as observed in the FDM analysis. [31][32][33] The results indicate that meso-erythritol crystallizes as a stable-form anhydride in the freeze-drying process. Absence of the metastable form and/or hydrate crystals is favorable to prevent physical changes of the excipient or chemical degradation of co-lyophilized APIs during storage.…”

Section: Discussionmentioning

confidence: 99%

Physical Characterization of meso-Erythritol as a Crystalline Bulking Agent for Freeze-Dried Formulations

Fujii

Izutsu²,

Kume

et al. 2015

CHEMICAL & PHARMACEUTICAL BULLETIN

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The purpose of this study was to identify and characterize new crystalline bulking agents applicable to freeze-dried pharmaceuticals. Thermal analysis of heat-melt sugar and sugar alcohol solids as well as their frozen aqueous solutions showed high crystallization propensity of meso-erythritol and D-mannitol. Experimental freeze-drying of the aqueous meso-erythritol solutions after their cooling by two different methods (shelf-ramp cooling and immersion of vials into liquid nitrogen) resulted in cylindrical crystalline solids that varied in appearance and microscopic structure. Powder X-ray diffraction and thermal analysis indicated different crystallization processes of meso-erythritol depending on the extent of cooling. Cooling of the frozen meso-erythritol solutions at temperatures lower than their T g ′ (glass transition temperature of maximally freeze-concentrated phase, 59.7°C) induced a greater number of nuclei in the highly concentrated solute phase. Growth of multiple meso-erythritol anhydride crystals at around 40°C explains the powder-like fine surface texture of the solids dried after their immersion in liquid nitrogen. Contrarily, shelf-ramp cooling of the frozen solution down to 40°C induced an extensive growth of the solute crystal from a small number of nuclei, leading to scale-like patterns in the dried solids. An early transition of the freezing step into primary drying induced collapse of the non-crystalline region in the cakes. Appropriate process control should enable the use of meso-erythritol as an alternative crystalline bulking agent in freeze-dried formulations.Key words meso-erythritol; crystallization; freeze-drying; sugar alcohol; thermal analysis Freeze-drying is a popular method for formulating various therapeutic proteins, liposomes, and antibiotics that are not sufficiently stable during storage in aqueous solutions or during drying at high temperatures.1,2) The process is also used to prepare amorphous solid formulations that foster better dissolution rates and bioavailability of poorly soluble pharmaceuticals. Various active pharmaceutical ingredients (APIs) and excipients composing the freeze-dried solids are either in their amorphous or crystalline states.3) The component crystallinity is determined by various factors including their intrinsic properties, co-solute compositions, and thermal history of the process. 4)Many freeze-dried formulations contain amorphous or crystalline bulking agents to prevent loss of the APIs during handling and/or provide better appearance. Some disaccharides (e.g., sucrose and trehalose) form glass-state amorphous dried solids that protect the conformation of embedded proteins and supramolecular assembly of liposomes from dehydration-induced irreversible conformational changes during the process and from their chemical degradation during storage.5,6) Some excipients (e.g., D-mannitol and glycine) are used to obtain superior appearance of stable crystalline solids, although they do not show protein-stabilizing effects. 7) They crystallize mainly ...

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Freeze-Dry Microscopy of Protein/Sugar Mixtures: Drying Behavior, Interpretation of Collapse Temperatures and a Comparison to Corresponding Glass Transition Data

Cited by 99 publications

References 44 publications

Component Crystallization and Physical Collapse during Freeze-Drying of <small>L</small>-Arginine–Citric Acid Mixtures

Component Crystallization and Physical Collapse during Freeze-Drying of <small>L</small>-Arginine–Citric Acid Mixtures

Design of the Lyophilization Process of a Doxorubicin Formulation Based on Thermal Properties

Physical Characterization of <i>meso</i>-Erythritol as a Crystalline Bulking Agent for Freeze-Dried Formulations

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