title: Evaluation of etanercept stability as exposed to various sugars with biophysical assessment.
abstract: Even though sugars have been used widely as additives for protein formulations, their exact mechanisms of protein stabilization and applicability remain still in need of investigation. The main purpose of this study was to evaluate the effects of various sugars on the biophysical stability of etanercept (Enbrel(®)). Six well known sugars including glucose, fructose, maltose, sucrose, trehalose, and raffinose were incorporated into the protein solution with different concentrations. The samples were analyzed with dynamic light scattering (DLS), differential scanning calorimetry (DSC), circular dichroism (CD), and size-exclusion chromatography (SEC). The DLS measurement showed that as the number of simple sugars and solution concentration increased, the hydrodynamic size increased with a decreasing absolute zeta potential. The DSC result provided consistent trends with the DLS data. As the concentration of sugar increased, the protein transition temperature (T(m)) was gradually increased in most of samples. In addition, a non-enzymatic browning reaction (NEB) was observed during heating of the sugar solution. To monitor the storage stability, sample solutions were stored at 4 and 40 °C. At 4 °C, the ratio of monomer, aggregate, and fragment were not significantly changed. However, fragmentation of etanercept was observed in accelerated storage. In addition, fructose and maltose showed a peak shift in the SEC result. Those results suggest that the reducing ability of sugar might be a reason for the different etanercept degradation pathways. Therefore, sugars need to be carefully considered to achieve the maximum efficiency of therapeutic proteins for the development of protein formulations.
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea.
authors.forename: Dae Gon
authors.initials: DG
authors.lastname: Lim
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea.
authors.forename: Nam Ah
authors.initials: NA
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea.
authors.forename: Jun Yeul
authors.initials: JY
authors.lastname: Lim
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea.
authors.forename: Ki Hyun
authors.initials: KH
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea.
authors.forename: Shavron
authors.initials: S
authors.lastname: Hada
authors.affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang, Gyeonggi 410-820, South Korea. Electronic address: shjeong@dongguk.edu.
authors.forename: Seong Hoon
authors.initials: SH
authors.lastname: Jeong
mesh_terms: D018501:Antirheumatic Agents / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002241:Carbohydrates / Q000737:chemistry*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004355:Drug Stability; D004356:Drug Storage; D000068800:Etanercept; D007074:Immunoglobulin G / Q000737:chemistry*; D018124:Receptors, Tumor Necrosis Factor / Q000737:chemistry*; D013696:Temperature; D044366:Transition Temperature
chemical_list: D018501:Antirheumatic Agents; D002241:Carbohydrates; D007074:Immunoglobulin G; D018124:Receptors, Tumor Necrosis Factor; D000068800:Etanercept
keywords: Differential scanning calorimetry; Etanercept; Fructose; Raffinose; Sugar