一种rAAV冻液制剂处方筛选的研究

Study on the Pre scription Screening of rAAV Jelly PreparationFan Xindan1,2,Li Dongjiao1,Zhang Min2,Wu Hao1(1.Shenyang Pharmaceutical University, School of Pharmaceutical Engineering, Shenyang, Liaoning, 110000, China；2.Neopharm (Suzhou) Biotechnology Co., Ltd., Suzhou, Jiangsu, 215000, China)

关键词:  rAAV2  处方筛选  质量属性  稳定性研究

Key words:  rAAV2  prescription screening  quality attribute  stability study

摘要:

目的 腺相关病毒(adeno-associated virus,AAV)以其安全性和高效性,成为基因治疗产品的重要载体。现在有大量的基于AAV的基因治疗产品处于临床阶段,并且已经有8款基于重组AAV载体的药物被美国食品药品监督管理局(Food and Drug Administration,FDA)和欧洲药品管理局(European Medicines Agency,EMA)批准上市。用于基因治疗的AAV产品具有较高的病毒基因组滴度导致AAV在制剂中容易发生聚集,使药物失去活性。方法 本研究考察一种基于血清型2型重组腺相关病毒rAAV2的基因治疗产品在不同处方和不同应力条件下(25℃±2℃,5次冻融循环和光照)稳定性,通过评估样品的关键质量属性包括外观、病毒基因组滴度、实心率、纯度、比滴度等,筛选出较优处方。结果 稳定性研究结果显示处方10 mM Tris缓冲液、180 mM氯化钠、0.001 wt%泊洛沙姆188,pH 7.5表现最优。结论 本研究讨论了rAAV2基因治疗产品处方开发的过程和结果,为相似rAAV产品的处方开发提供了参考。

Abstract:

OBJECTIVE Adeno-associated virus (AAV) has emerged as a pivotal vector for gene therapy products due to its safety and high efficiency. Currently, numerous AAV-based gene therapy products are in clinical stages, with eight drugs based on recombinant AAV vectors approved by the Food and Drug Administration (FDA) and European Medicines Agency (EMA). Gene therapy products utilizing AAV typically have a high viral genome titer, which can lead to aggregation in the formulation, rendering the drug inactive. METHODS This study investigates the stability of a gene therapy product based on serotype 2 recombinant adeno-associated virus (rAAV2) under various prescriptions and stress conditions (25 °C ±2 °C, 5 freeze-thaw cycles, and light exposure). We identified the optimal prescription by evaluating key quality attributes of the samples, including appearance, viral genome titer, solid rate, purity, and specific titer. RESULTS The stability study results indicated that a formulation containing 10 mM Tris buffer, 180 mM sodium chloride, 0.001 wt% Poloxamer 188, and pH 7.5 exhibited the best performance. CONCLUSION This paper details the process and results of the prescription development for rAAV2 gene therapy products and offers a reference for the development of similar rAAV products.