title: Long-term stabilization of a new freeze-dried and albumin-free formulation of recombinant human interferon alpha 2b.
abstract: In this work, we evaluate the stability of a new freeze-dried and albumin-free formulation of recombinant human IFN alpha 2b (rhIFN-alpha2b) to be used in humans. The freeze-dried, albumin-free formulation was stored at the recommended temperature of 4 degrees C, and under accelerated storage conditions (28 degrees C). The stability of this product was also compared with the stability of a liquid albumin-free formulation of this cytokine. Finally, the stability of the freeze-dried albumin-free formulation was examined after reconstitution and storage at 4 degrees C and room temperature (28 degrees C) for 30 days. Samples were periodically subjected to biological activity assay (antiviral titration), reversed phase high-performance liquid chromatography (RP-HPLC), pyrogens, sterility and enzyme-linked immunosorbent assay (ELISA) testing, abnormal toxicity screening, organoleptic evaluation, and measurement of residual moisture and pH. Accelerated storage (28 degrees C) data for the freeze-dried albumin-free formulation showed biochemical stability of the active ingredient throughout the 6-month study, showing activity between 85 and 125% of its nominal value. RP-HPLC-determined purity showed that rhIFN-alpha2b remained above 95%. Additionally, the formulation was non-pyrogenic, non-toxic, sterile, and organoleptically acceptable. The real-time storage data confirmed the good biochemical long-term (30 months) stability of the freeze-dried formulation of this cytokine. Comparison with the liquid rhIFN-alpha2b albumin-free preparation showed that the freeze-dried albumin-free formulation maintained the stability of the active ingredient better than the liquid preparation. The formulation was also stable after reconstitution and storage at 4 degrees C and 28 degrees C, for 30 days.
authors.affiliation: Department of Development and Formulations, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana, Cuba.
authors.forename: Llamil
authors.initials: L
authors.lastname: Ruiz
authors.forename: Nuria
authors.initials: N
authors.lastname: Reyes
authors.forename: Jorge
authors.initials: J
authors.lastname: Sotolongo
authors.forename: Kethia
authors.initials: K
authors.lastname: Aroche
authors.forename: Raymersy
authors.initials: R
authors.lastname: Aldana
authors.forename: Reynier
authors.initials: R
authors.lastname: Báez
authors.forename: Eugenio
authors.initials: E
authors.lastname: Hardy
mesh_terms: D000418:Albumins / Q000032:analysis; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology; D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D004355:Drug Stability; D004356:Drug Storage; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005612:Freeze Drying; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000737:chemistry* / Q000494:pharmacology; D008574:Mengovirus / Q000187:drug effects / Q000502:physiology; D011994:Recombinant Proteins; D013696:Temperature; D013997:Time Factors
chemical_list: D000418:Albumins; D000970:Antineoplastic Agents; D000998:Antiviral Agents; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins