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D20A0004

Development of Novel Polysaccharide Nanoparticles as Vaccine Nano-carrier and Adjuvant

Invention Description
In this invention, we have in vitro and in vivo verified that Fucoidan (FUC)/quaternary chitosan (qCS) nanoparticles (FqC-NPs) are good nano-carriers and adjuvant candidates for intraperitonially administered anthrax vaccine adsorbed (AVA). Positively ( ) and negatively (–) surface-charged FqC-NPs were facile prepared via polyelectrolyte complexation by varying the mass ratio of FUC and qCS (such as trimethyl chitosan and N-[(2-hydroxy-3-trimethylammonium)propyl]chitosan). Both charged NP types permitted high viability and presented no cytotoxicity on L929 cells, A549 cells and JAWS II dendritic cells (DC). They also showed no plasma membrane damage, observed via lactate dehydrogenase assays, and excellent uptake efficacy in JAWS II dendritic cells. In vitro intracellular cytokine flow cytometry studies showed that higher levels of pro-inflammatory cytokines produced by JAWS II DCs and could therefore serve as an adjuvant to enhance immunity without inducing immunotoxicity. In vivo studies on A/J mice showed that when challenged with the anthrax toxin, ( )-FqC-NPs carrying AVA triggered a more efficient response with a higher IgG anti-PA antibody titer, and yielded higher survival rates than AVA with CpG oligodeoxynucleotides (CpG ODNs), or AVA alone. Our data collectively suggest that the cheaper FqC-NPs may be a promising adjuvant for use with the anthrax vaccine or with other infectious diseases, and can be used instead of CpG.

Competitive Advantages
For the clinical applications, NP-based adjuvants have potential limitations due to their difficult preparation, unacceptable toxicity, insufficient efficacy, and high economic costs. In this study, we proved that FqC-NPs exhibited no cytotoxicity and immunotoxicity toward DCs, and in vitro cellular uptake studies revealed that FqC-NPs showed excellent uptake efficacy in JAWS II DCs. In terms of their preparation, FqC-NPs were easily prepared with high reproductivity via electrostatic interactions between two opposite-charged polyelectrolytes of a simple mixing process. Furthermore, FqC-NPs are more cost-effective than CpG ODNs to reduce the price of vaccines.

Intellectual Property
US Patent (16/392,856) and ROC Patent (108129610) are pending.

Key Publication
1. CC Chuang, MH Tsai, HJ Yen, HF Shyu, KM Cheng, XA Chen, CC Chen, JJ Young, JH Kau, “A fucoidanquaternary chitosan nanoparticle adjuvant for anthrax vaccineas an alternative to CpG oligodeoxynucleotides” Carbohydrate Polymers 229 (2020) 1154032.
2. MH Tsai, CC Chuang, CC Chen, HJ Yen, KM Cheng, XA Chen, HF Shyu, CY Lee, JJ Young, JH Kau, “Nanoparticles assembled from fucoidan and trimethylchitosan as anthrax vaccine adjuvant: In vitro and in vivo efficacy in comparison to CpG” Carbohydrate Polymers 236 (2020) 116041.


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