Benington Leah R, Rajan Gunesh, Locher Cornelia, Lim Lee Yong
Division of Pharmacy, School of Allied Health, University of Western Australia, Perth 6009, Australia.
Otolaryngology, Head & Neck Surgery, Division of Surgery, Medical School, University of Western Australia, Perth 6009, Australia.
Pharmaceutics. 2021 Oct 21;13(11):1762. doi: 10.3390/pharmaceutics13111762.
Basic fibroblast growth factor (FGF-2) is a highly labile protein with strong potential for tissue engineering. The aim of this study was to develop FGF-2 formulations that are stable against physical stressors encountered in pharmaceutical processing and evaluation. Pharmaceutical excipients, alone or in combination, were added to aqueous FGF-2 (770 ng/mL) solution and the stability of the resulting solutions on storage at 4-37 °C was evaluated. Stability of the solutions to repeated freeze-thaw cycles and lyophilisation was also evaluated, as well as the stability of the lyophilised stabilised protein to storage at -4, 4 and 18 °C for up to 12 months. In all of these experiments FGF-2 was quantified by ELISA assay. The as-received FGF-2, when dissolved in water, was highly unstable, retaining only 50% of baseline protein content after 30 min at 37 °C or 1 h at 25 °C. By contrast, FGF-2 solutions prepared with 0.5% methylcellulose (MC) and 20 mM alanine (formulation F5) or with 0.5% MC and 1 mg/mL human serum albumin (HSA) (formulation F6) were highly stable, having residual FGF-2 content comparable to baseline levels even after 2 h at 37 °C and 5 h at 25 °C. F5 and F6 were also highly stable to repeated freeze-thaw cycles, with >99% of FGF-2 load remaining after the third cycle. In addition, F5 and F6 were stable to lyophilisation, and the lyophilised products could be stored at -4, 4 or 18 °C for at least 12 months, with less than 1% loss in mean FGF-2 content. Thus, FGF-2 solution is effectively stabilised against both thermal and processing stressors in the presence of MC and alanine (F5), or MC and HSA (F6). The resultant FGF-2 solutions may be applied as medicinal products or further processed into more advanced medicinal products, e.g., scaffolds, for wound healing and tissue regeneration.
碱性成纤维细胞生长因子（FGF - 2）是一种高度不稳定的蛋白质，在组织工程方面具有巨大潜力。本研究的目的是开发出在药物加工和评估过程中能抵抗物理应激因素的FGF - 2制剂。将药用辅料单独或组合添加到FGF - 2水溶液（770 ng/mL）中，并评估所得溶液在4 - 37°C储存时的稳定性。还评估了溶液对反复冻融循环和冻干的稳定性，以及冻干后的稳定化蛋白质在 - 4°C、4°C和18°C储存长达12个月的稳定性。在所有这些实验中，通过酶联免疫吸附测定（ELISA）法对FGF - 2进行定量。刚收到的FGF - 2溶解于水时极不稳定，在37°C下30分钟或25°C下1小时后，仅保留50%的基线蛋白质含量。相比之下，用0.5%甲基纤维素（MC）和20 mM丙氨酸制备的FGF - 2溶液（制剂F5）或用0.5% MC和1 mg/mL人血清白蛋白（HSA）制备的溶液（制剂F6）非常稳定，即使在37°C下2小时和25°C下5小时后，剩余的FGF - 2含量仍与基线水平相当。F5和F6对反复冻融循环也高度稳定，在第三次循环后仍有>99%的FGF - 2负载量。此外，F5和F6对冻干稳定，冻干产品可在 - 4°C、4°C或18°C下储存至少12个月，平均FGF - 2含量损失不到1%。因此，在MC和丙氨酸（F5）或MC和HSA（F6）存在的情况下，FGF - 2溶液能有效抵抗热应激和加工应激因素。所得的FGF - 2溶液可作为药品应用，或进一步加工成更先进的药品，如用于伤口愈合和组织再生的支架。
