苯甲醇对依那西普制剂稳定性的多因素研究及开发指导

A Multifactorial Study of Benzyl Alcohol Effects on Etanercept Formulation Stability and Development Guidance

关键词:  苯甲醇  依那西普  多剂量制剂  稳定性

Key words:  benzyl alcohol  etanercept  multi-dose formulations  stability

摘要:

目的 为筛选合适的苯甲醇(benzyl alcohol，BA)浓度，深入探究其在制剂多重应力条件下对稳定性的影响，以支持依那西普制剂长期储存。方法 以依那西普为模型药物，系统评估含BA多剂量制剂在低温、室温、高温、光照、冻融及振荡等多种应力条件下的稳定性。采用动态光散射(dynamic light scattering，DLS)、差示扫描量热法(differential scanning calorimetry，DSC)、纳米差示扫描荧光法(nano differential scanning fluorimetry，nanoDSF)、固有荧光(intrinsic fluorescence，IF)、尺寸排阻色谱(size exclusion chromatography，SEC)及8-苯胺基-1-萘磺酸等技术检测制剂稳定性，并通过抑菌实验评估其抗菌活性。结果 DLS与IF检测显示，BA在加入制剂的初始阶段即诱导依那西普构象展开。DSC与nanoDSF结果表明，依那西普的稳定性随BA浓度升高呈依赖性降低，更易发生聚集。多重应力条件下的SEC稳定性评估进一步证实，BA会诱导依那西普产生浓度依赖性降解，提示含BA的依那西普制剂需在2~4°C条件下保存的重要性。结论 本研究发现，在该研究框架内，0.9% BA为兼顾抗菌效力与聚集风险最小化的较优浓度。

Abstract:

OBJECTIVE To investigate the impact of benzyl alcohol (BA) concentration on formulation stability under multiple stress conditions, thereby supporting the long-term storage of etanercept. METHODS Using etanercept as a model drug, this study systematically evaluated the stability of BA-containing multi-dose formulations under various stress conditions, including low temperature, room temperature, high temperature, light exposure, freeze-thaw cycles, and shaking. Techniques such as dynamic light scattering (DLS), differential scanning calorimetry (DSC), nano differential scanning fluorimetry (nanoDSF), intrinsic fluorescence (IF), size exclusion chromatography (SEC), and 8-anilino-1-naphthalenesulfonic acid were employed to assess formulation stability, and antibacterial activity was evaluated through bacteriostatic experiments. RESULTS DLS and IF analyses showed that BA induced initial conformational unfolding of the drug immediately after addition to the formulation. DSC and nanoDSF results indicated that the stability of etanercept decreased in a BA concentration-dependent manner, making it more prone to aggregation. Stability assessments by SEC under multiple stress conditions further confirmed that BA induced concentration-dependent degradation of etanercept, highlighting the importance of storing BA-containing etanercept formulations at 2-4°C. CONCLUSION This study found that within the research framework, 0.9% BA represents the optimal concentration balancing antibacterial efficacy with minimized aggregation risk.