Preformulation Studies and Physicochemical Properties of Intranasal Low Sialic Acid Erythropoietin

by A. Muñoz-Cernada, PhD, J. Cardentey-Fernández, L. Paradina-Fernández, L. Rojas-del Calvo, T. Figueredo-Gonzáles, D. Rodríguez-Abreu, M. Fernández-Cervera, PhD, I. Sosa-Testé, PhD, and D. Amaro-González, PhD
Volume 15, Issue 2 (Summer 2016)

Intranasal low sialic acid erythropoietin (Neuro-EPO) is a non-hematopoietic molecule that shows neuro-protective activity in some rodent models of brain ischemia. The protein formulations are susceptible to a loss of stability due to formulation, processing, and storage. The aim of this study was to perform formulation analyses of Neuro-EPO to achieve acceptable stability. Experiments were done to assess the compatibility of Neuro-EPO with selected excipients: benzalkonium chloride (BAC) preservative, sodium chloride (NaCl), and hydroxypropyl methylcellulose (HPM) K4M polymer. Instabilities were induced by creating thermal stress conditions (25 ± 2°C). Protein degradation was measured using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)/Western blot, reversed-phase high-pressure liquid chromatography (RP-HPLC), and size-exclusion HPLC (SE-HPLC). Neuro-EPO excipient compatibility tests showed instabilities with BAC, which seemed to be related to protein aggregation. Non-preserved formulations maintained a biological activity similar to that observed with the formulation preserved...

Citation:
Muñoz-Cernada A, Cardentey-Fernández J, Paradina-Fernández L, Rojas-del Calvo L, Figueredo-Gonzáles T, Rodríguez-Abreu D, Fernández-Cervera M, Sosa-Testé I, Amaro-González D. Preformulation studies and physicochemical properties of intranasal low sialic acid erythropoietin. BioProcess J, 2016; 15(2): 45–51. http://dx.doi.org/10.12665/J152.Amaro.

Posted online July 30, 2016.