Quality of bevacizumab (Avastin®) repacked in single-use glass vials for intravitreal administration

ABSTRACT Purpose: Avastin® (bevacizumab) is an anti-vascular endothelial growth factor (VEGF) monoclonal antibody given as an off-label drug by intravitreal administration for treatment of ocular diseases. The drug's clinical application and its cost-benefit profile has generated demand for its division into single-use vials to meet the low volume and low-cost doses necessary for intraocular administration. However, the safety of compounding the drug in single-use vials is still under discussion. In this study, the stability and efficacy of Avastin® repacked in individual single-use glass vials and glass ampoules by external compounding pharmacies were evaluated. Methods: Polyacrylamide gel electrophoresis (PAGE), size-exclusion chromatography (SEC), dynamic light scattering (DLS), and turbidimetry were selected to detect the formation of aggregates of various sizes. Changes in bevacizumab biological efficacy were investigated by using an enzyme-linked immunosorbent assay (ELISA). Results: Repacked and reference bevacizumab showed similar results when analyzed by PAGE. By SEC, a slight increase in high molecular weight aggregates and a reduction in bevacizumab monomers were observed in the products of the three compounding pharmacies relative to those in the reference bevacizumab. A comparison of repacked and reference SEC chromatograms showed that the mean monomer loss was ≤1% for all compounding pharmacies. Protein aggregates in the nanometer- and micrometer-size ranges were not detected by DLS and turbidimetry. In the efficacy assay, the biological function of repacked bevacizumab was preserved, with <3% loss of VEGF binding capacity relative to that of the reference. Conclusion: The results showed that bevacizumab remained stable after compounding in ampoules and single-use glass vials; no significant aggregation, fragmentation, or loss of biological activity was observed.

摘要
目的：阿瓦斯汀®（Avastin®，bevacizumab，贝伐珠单抗）是一种抗血管内皮生长因子（vascular endothelial growth factor, VEGF）单克隆抗体，常作为超说明书用药通过玻璃体内注射给药用于眼部疾病的治疗。该药物的临床应用及其成本效益特征催生了将其分装为单次使用小瓶的需求，以满足眼内给药所需的低给药体积、低成本给药剂量的要求。然而，将该药物调配分装至单次使用小瓶的安全性仍存在争议。本研究评估了由外部药品调配药房分装至独立单次使用玻璃小瓶与玻璃安瓿瓶中的阿瓦斯汀®的稳定性与生物学活性。
方法：本研究选用聚丙烯酰胺凝胶电泳（polyacrylamide gel electrophoresis, PAGE）、尺寸排阻色谱法（size-exclusion chromatography, SEC）、动态光散射（dynamic light scattering, DLS）及浊度法，检测不同尺寸的蛋白质聚集体的形成情况；采用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）探究贝伐珠单抗生物学活性的变化。
结果：经聚丙烯酰胺凝胶电泳分析，重新分装的贝伐珠单抗与参比制剂结果相近。尺寸排阻色谱结果显示，相较于参比制剂，三家调配药房生产的产品中高分子量聚集体略有升高，贝伐珠单抗单体占比有所降低；对重新分装与参比制剂的尺寸排阻色谱图进行对比后发现，所有调配药房的产品单体平均损失率均≤1%。动态光散射与浊度法均未检测到纳米级及微米级的蛋白质聚集体。活性检测结果显示，重新分装的贝伐珠单抗保留了生物学功能，其血管内皮生长因子结合能力相较于参比制剂的损失率＜3%。
结论：本研究结果表明，贝伐珠单抗经调配分装至安瓿瓶与单次使用玻璃小瓶后仍保持稳定，未观察到明显的聚集、降解或生物学活性损失。