Zhang Xinyue, Su Guangpu, Shao Zitong, Chan Ho Wan, Li Si, Chow Stephanie, Tsang Chi Kwan, Chow Shing Fung
Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, L2-08B, 2/F, Laboratory Block, 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
Advanced Biomedical Instrumentation Centre, Hong Kong Science Park, Shatin, Hong Kong SAR, China.
Drug Deliv Transl Res. 2025 Jun;15(6):2022-2047. doi: 10.1007/s13346-024-01721-8. Epub 2024 Nov 1.
Ischemic stroke is one of the major diseases causing varying degrees of dysfunction and disability worldwide. The current management of ischemic stroke poses significant challenges due to short therapeutic windows and limited efficacy, highlighting the pressing need for novel neuroprotective treatment strategies. Previous studies have shown that fingolimod (FIN) is a promising neuroprotective drug. Here, we report the rational development of FIN nano-embedded nasal powders using full factorial design experiments, aiming to provide rapid neuroprotection after ischemic stroke. Flash nanoprecipitation was employed to produce FIN nanosuspensions with the aid of polyvinylpyrrolidone and cholesterol as stabilizers. The optimized nanosuspension (particle size = 134.0 ± 0.6 nm, PDI = 0.179 ± 0.021, physical stability = 72 ± 0 h, and encapsulation efficiency of FIN = 90.67 ± 0.08%) was subsequently spray-dried into a dry powder, which exhibited excellent redispersibility (RdI = 1.09 ± 0.04) and satisfactory drug deposition in the olfactory region using a customized 3D-printed nasal cast (45.4%) and an Alberta Idealized Nasal Inlet model (8.6%) at 15 L/min. The safety of the optimized FIN nano-embedded dry powder was confirmed in cytotoxicity studies with nasal (RPMI 2650 and Calu-3 cells) and brain related cells (SH-SY5Y and PC 12 cells), while the neuroprotective effects were demonstrated by observed behavioral improvements and reduced cerebral infarct size in a middle cerebral artery occlusion mouse stroke model. The neuroprotective effect was further evidenced by increased expression of anti-apoptotic protein BCL-2 and decreased expression of pro-apoptotic proteins CC3 and BAX in brain peri-infarct tissues. Our findings highlight the potential of nasal delivery of FIN nano-embedded dry powder as a rapid neuroprotective treatment strategy for acute ischemic stroke.
缺血性中风是全球范围内导致不同程度功能障碍和残疾的主要疾病之一。由于治疗窗口期短和疗效有限，目前缺血性中风的治疗面临重大挑战，这凸显了对新型神经保护治疗策略的迫切需求。先前的研究表明，芬戈莫德（FIN）是一种有前景的神经保护药物。在此，我们报告了使用全因子设计实验合理开发FIN纳米包埋鼻用粉剂，旨在为缺血性中风后提供快速神经保护。借助聚乙烯吡咯烷酮和胆固醇作为稳定剂，采用快速纳米沉淀法制备FIN纳米混悬液。随后将优化后的纳米混悬液（粒径 = 134.0 ± 0.6 nm，多分散指数 = 0.179 ± 0.021，物理稳定性 = 72 ± 0 h，FIN包封率 = 90.67 ± 0.08%）喷雾干燥成干粉，使用定制的3D打印鼻模（45.4%）和艾伯塔理想化鼻入口模型（8.6%）在15 L/min时，该干粉表现出优异的再分散性（再分散指数 = 1.09 ± 0.04）和在嗅觉区域令人满意的药物沉积。在对鼻相关细胞（RPMI 2650和Calu - 3细胞）和脑相关细胞（SH - SY5Y和PC 12细胞）的细胞毒性研究中证实了优化后的FIN纳米包埋干粉的安全性，而在大脑中动脉闭塞小鼠中风模型中，通过观察到的行为改善和脑梗死体积减小证明了其神经保护作用。脑梗死周围组织中抗凋亡蛋白BCL - 2表达增加和促凋亡蛋白CC3及BAX表达降低进一步证明了其神经保护作用。我们的研究结果突出了鼻内递送FIN纳米包埋干粉作为急性缺血性中风快速神经保护治疗策略的潜力。
