Effect of Mannitol Crystallization on the Stability and Aerosol Performance of a Spray-Dried Pharmaceutical Protein, Recombinant Humanized anti-IgE Monoclonal Antibody

doi:10.1021/js9800679

Journal of Pharmaceutical Sciences

Volume 87, Issue 11, November 1998, Pages 1406-1411

https://doi.org/10.1021/js9800679 Get rights and content

Abstract

We have examined the stability and aerosol performance of the pharmaceutical protein recombinant humanized anti-IgE mono-clonal antibody (rhuMAbE25) spray dried with mannitol. The aerosol performance was measured by the fine particle fraction (FPF), and stability was assessed by the formation of soluble aggregates. When mannitol was added to the spray-dried rhuMAbE25 formulation, its ability to stabilize the protein leveled off above about 20% (w/w, dry basis). The FPF of the spray-dried formulations was stable during storage for rhuMAbE25 containing 10% and 20% mannitol, but the 30% formulation exhibited a dramatic decrease upon storage at both 5 °C and 30 °C, due to mannitol crystallization. We tested the addition of sodium phosphate to a 60:40 rhuMAbE25:mannitol (w:w) mixture, which otherwise crystallized upon spray drying and yielded a nonrespirable powder. The presence of sodium phosphate was successful in inhibiting mannitol crystallization upon spray drying and dramatically lowering the rate of solid-state aggregation. However, over long-term storage some crystallization was observed even for the phosphate-containing samples, concomitantly with increased particle size and decreased suitability for aerosol delivery. Therefore, the physical state of mannitol (i.e., amorphous or crystalline) plays a role both in maintaining protein stability and providing suitable aerosol performance when used as an excipient for spray-dried powders. Agents which retard mannitol crystallization, e.g., sodium phosphate, may be useful in extending the utility of mannitol as an excipient in spray-dried protein formulations.

References and Notes (22)

Y.-F. Maa et al.
Spray drying of air-liquid interface sensitive recombinant human growth hormone
J. Pharm. Sci.
(1998)
J. Heyder et al.
Deposition of particles in the human respiratory tract in the size range of 0.005-15 µm
J. Aerosol Sci.
(1986)
H.R. Costantino et al.
Solid-phase aggregation of proteins under pharmaceutically relevant conditions
J. Pharm. Sci.
(1994)
B.C. Hancock et al.
Characteristics and significance of the amorphous state in pharmaceutical systems
J. Pharm. Sci.
(1997)
J. Broadhead et al.
The effect of process and formulation variables on the properties of spray-dried β-galactosidase
J. Pharm. Phar-macol.
(1994)
M. Mumenthaler et al.
Feasibility study on spray-drying protein pharmaceuticals: recombinant human growth hormone and tissue-type plasminogen activator
Pharm. Res.
(1994)
H.-K. Chan et al.
Spray dried powders and powder blends of recombinant human deoxyribonuclease (rhDNase) for aerosol delivery
Pharm. Res.
(1997)
Y.-F. Maa et al.
The effect of operating and formulation variables on the morphology of spray-dried protein particles
Pharm. Dev. Technol.
(1997)
H.R. Costantino et al.
Fourier transform infrared spectroscopic analysis of the secondary structure of recombinant humanized immunoglo-bulin G
Pharm. Sci.
(1997)
Y.-F. Maa et al.
Effect of spray drying and subsequent processing conditions on residual moisture content and physical/biochemical stability of protein inhalation powders
Pharm. Res.
(1998)

M.J. Hageman

Water sorption and solid-state stability of proteins

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Citation Excerpt :
Even with the differences in surface areas, the correlation between the surface area (Fig. 10-B) or particle size distribution with the physical stability of the formulations was not observed in this study. Presence of crystallinity within the dried formulation has been known to cause protein instability because it leads to phase separation of the protein and excipient in the solid matrix.34,35 The impact of crystallization on the stability of SFD formulations has been previously studied17 For both processes in this study, the formulations with 15 mg/ml and 20 mg/ml of mannitol showed the presence of crystallinity upon drying with the rest of the formulations staying amorphous (Fig. 5).
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Conference ReportsEffect of Mannitol Crystallization on the Stability and Aerosol Performance of a Spray-Dried Pharmaceutical Protein, Recombinant Humanized anti-IgE Monoclonal Antibody

Abstract

J. Pharm. Sci.

J. Aerosol Sci.

J. Pharm. Sci.

J. Pharm. Sci.

The effect of process and formulation variables on the properties of spray-dried β-galactosidase

J. Pharm. Phar-macol.

Feasibility study on spray-drying protein pharmaceuticals: recombinant human growth hormone and tissue-type plasminogen activator

Pharm. Res.

Spray dried powders and powder blends of recombinant human deoxyribonuclease (rhDNase) for aerosol delivery

Pharm. Res.

The effect of operating and formulation variables on the morphology of spray-dried protein particles

Pharm. Dev. Technol.

Fourier transform infrared spectroscopic analysis of the secondary structure of recombinant humanized immunoglo-bulin G

Pharm. Sci.

Effect of spray drying and subsequent processing conditions on residual moisture content and physical/biochemical stability of protein inhalation powders

Pharm. Res.

Water sorption and solid-state stability of proteins

Conference Reports
Effect of Mannitol Crystallization on the Stability and Aerosol Performance of a Spray-Dried Pharmaceutical Protein, Recombinant Humanized anti-IgE Monoclonal Antibody