Effects of annealing lyophilized and spray-lyophilized formulations of recombinant human interferon - Surfactants

Effects of annealing lyophilized and spray-lyophilized formulations of recombinant human interferon
Published Online: 21 Jan 2003
Received: 29 April 2002; Revised: 13 September 2002; Accepted: 22 October 2002
Serena D. Webb 1, Jeffrey L. Cleland 2, John F. Carpenter 3, Theodore W. Randolph 1 *
Journal of Pharmaceutical Sciences
Volume 92, Issue 4 , Pages 715 - 729
Wiley InterScience
1Department of Chemical Engineering, University of Colorado, Department of Engineering, Center for Pharmaceutical Biotechnology, Engineering Center, Room ECCH 111, Boulder, Colorado 80309-0424
2Genentech, Inc., South San Francisco, California 94080
3Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, Colorado 80262

email: Theodore W. Randolph (Randolph@pressure3.colorado.edu)
*Correspondence to Theodore W. Randolph, Department of Chemical Engineering, University of Colorado, Department of Engineering, Center for Pharmaceutical Biotechnology, Engineering Center, Room ECCH 111, Boulder, Colorado 80309-0424. Telephone: 303-492-4776; Fax: 303-492-4341
Keywords
rhIFN- ? surfactant, Tween 20 ? polysorbate 20 ? lyophilization ? freeze drying ? spray-lyophilization ? annealing ? ice/liquid interface ? air/liquid interface
Abstract
The purpose of this study was to examine the effects of adsorption of recombinant human interferon- (rhIFN-) on ice surfaces and subsequent drying during processing by spray-lyophilization and lyophilization. Ice/liquid interfacial areas were manipulated by the freezing method as well as by the addition of an annealing step during lyophilization; that is, rhIFN- adsorption was modified by the addition of nonionic surfactants. rhIFN- was lyophilized or spray-lyophilized at a concentration of 1 mg/mL in 5% sucrose, 5% hydroxyethyl starch (HES) ? 0.03% polysorbate 20 in 140 mM KCl, and 10 mM potassium phosphate, pH 7.5. After the samples were frozen, half were annealed on the lyophilizer shelf. Recovery of soluble protein was measured at intermediate points during processing. On drying, the secondary structure of rhIFN- was determined by second-derivative infrared (IR) spectroscopy, specific surface areas (SSAs) were measured, scanning electron micrographs (SEM) were taken, and dissolution times were recorded. Adsorption of rhIFN- to ice/liquid interfaces alone was not responsible for aggregation. Rather, drying was necessary to cause aggregation in lyophilized sucrose formulations. Addition of an annealing step to the lyophilization cycle resulted in more native-like secondary protein structure in the dried solid, eliminated cracking of the dried cakes, and suppressed both the formation of air/liquid interfaces and rhIFN- aggregation on reconstitution. ? 2003 Wiley-Liss, Inc. and the American pharmaceutical Association J Pharm Sci 92:715-729, 2003