Rational Development of Fingolimod Nano-embedded Microparticles as Nose-to-Brain Neuroprotective Therapy for Ischemic Stroke
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, leading to a 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 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 and an Alberta Idealized Nasal Inlet model. The safety of the optimized FIN dry powder was confirmed in cytotoxicity studies with nasal and brain cells, while the neuroprotective effects were demonstrated by observed behavioral improvements and reduced cerebral infarct size in an established 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.
Download the full article preprint, (not peer-reviewed yet) as PDF here: Rational Development of Fingolimod Nano-embedded Microparticles as Nose-to-Brain Neuroprotective Therapy for Ischemic Stroke
or read here
Materials
Fingolimod (FIN, > 99.9% purity) was purchased from Hefei Hirisun Phamatech (Hefei, China). Curcumin (CUR, > 99.5% purity) was sourced from Yung-Zip Chemicals (Taichung, Taiwan). Methanol (MeOH) and ethanol (EtOH) were obtained from VWR BDH Chemicals (VWR International S.A.S., Fontenay-sous-Bois, France). Ultra-purified water (UPW) was generated using a Barnstead NANOpure Diamond system (Thermo Fisher Scientific, Waltham, MA, USA). Polyvinylpyrrolidone (PVP K30) and cholesterol (CLT) were procured from Sigma-Aldrich (St. Louis, MO, USA), and Mannitol (Pearlitol 160C) was purchased from Roquette (Lestrem, France). Trifluoroacetic acid (TFA), Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12), Dulbecco’s Modified Eagle’s Medium (DMEM), Minimum Essential Medium (MEM), Kaighn’s Modification of Ham’s F-12 Medium (F-12K medium), 0.25% (w/v) trypsin-EDTA, phosphate-buffered saline (PBS, 10×), fetal bovine serum (FBS), and antibiotic–antimycotic (100×) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Additionally, a variety of cell lines were obtained from the American Type Cultural Collection (ATCC; Manassas, VA, USA), including PC 12 cells, RPMI 2650 cells, Calu-3 cells, and SH-SY5Y cells. Nissl Staining Solution (Cat. No. C0117) was purchased from Shanghai Beyotime Biotechnology Co., Ltd (Shanghai, China). Polyvinylidene difluoride membrane was purchased from Bio-Rad (California, USA). Primary antibodies against Cleaved Caspase-3 (CC3) (Cat. No. 9664) and β-Tubulin proteins (Cat. No. #2146S) were purchased from Cell Signaling Technology (Massachusetts, USA) whose t against B-cell lymphoma 2 (BCL-2) (Cat. No. WL01556) and BCL-2 associated X (BAX) (Cat. No. WL01637) proteins were purchased from Wanlei Bio (Liaoning, China). Horseradish peroxidase-labeled goat anti-rabbit IgG(H + L) secondary antibodies (Cat. No. A0208) were purchased from Beyotime (Shanghai, China).
Xinyue Zhang, Guangpu Su, Zitong Shao, Ho Wan Chan, Si Li, Stephanie Chow, Chi Kwan Tsang, Shing Fung Chow, Rational Development of Fingolimod Nano-embedded Microparticles as Nose-to-Brain Neuroprotective Therapy for Ischemic Stroke, https://doi.org/10.21203/rs.3.rs-4715108/v1, This work is licensed under a CC BY 4.0 License
See our next webinar:
“Controlled Release for Nutraceuticals“
Date: 10th of September, Time: 5:00 PM (Amsterdam, Berlin)
