Research Article – Pharmaceutical Nanotechnology
Tumor-Targeted Paclitaxel Delivery and Enhanced Penetration Using TAT-Decorated Liposomes Comprising Redox-Responsive Poly(Ethylene Glycol)

https://doi.org/10.1002/jps.24291Get rights and content

ABSTRACT

To combine the advantage of poly(ethylene gylcol) (PEG) for longer circulation and cell-penetrating peptides (CPPs) for efficient cellular uptake, paclitaxel (PTX)-loaded liposomes functionalized with TAT, the most frequently used CPP, and cleavable PEG via a redox-responsive disulfide linker (PTX-C-TAT-LP) were successfully developed here. Under physiological conditions, TAT was shielded by PEG layer and liposomes exhibited a long blood circulation. At tumor site, PEG could be detached in the presence of exogenous reducing agent [glutathione (GSH)] and TAT was exposed to facilitate cell internalization. In the presence of GSH, the liposomal vesicle C-TAT-LP showed increased cellular uptake and improved three-dimensional tumor spheroids penetration in vitro compared with analogous stable shielded liposomes. C-TAT-LP achieved enhanced tumor distribution and demonstrated superior delivery efficiency in vivo. PTX-C-TAT-LP with GSH strongly inhibited the proliferation of murine melanoma B16F1 tumor cells in vitro and in vivo with the tumor inhibition rate being 69.4% on B16F1-bearing mice. In addition, the serum aspartate transaminase level, alanine transaminase level, and creatine kinase level were almost completely within normal range in the PTX-C-TAT-LP with GSH group, revealing PTX-C-TAT-LP with GSH had no obvious drug-related adverse events for liver and heart. Taken together, C-TAT-LP is a promising tumor-targeting drug carrier. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Section snippets

INTRODUCTION

Nanotechnology-based targeted drug delivery systems have drawn much attention for improved efficacy and reduced toxicity by reducing side effects resulted from non-specific tissue distribution of free therapeutic molecules.1 To date, many nanocarriers have been prepared, such as micelles,2., 3. nanoparticles,4., 5. liposomes,6., 7. and so on. Liposomes have shown promising results in clinical and preclinical studies such as Doxil8 and DaunoXome,9 owing to their good biocompatibility, ease of

Materials

Paclitaxel was purchased from AP Pharmaceutical Company, Ltd. (Chongqing, China). Soybean phosphatidylcholine (SPC) was purchased from Shanghai Taiwei Chemical Company (Shanghai, China). Cholesterol (CHO) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Chengdu Kelong Chemical Company (Chengdu, China). Reduced GSH was purchased from Merck & Company, Inc. (Whitehouse Station, New Jersey).

Characterization of Liposomes

All liposomes exhibited a mean particle size between 100.43 and 113.10 nm and an average polydispersity index between 0.215 and 0.270 (Table 2), being adequate for a passive targeting based on EPR effect as the effective drug delivery using liposomes by EPR effect is highly dependent on size ranging from 100 to 200 nm in diameter.42 Compared with the positive potential of PTX-TAT-LP, PTX-C-TAT-LP showed a negative zeta potential (Table 2) because of the comodification of cleavable PEG on the

CONCLUSIONS

In our study, the PTX-loaded exogenous-GSH triggered TAT-presenting liposomes (PTX-C-TAT-LP) were constructed. Owing to the cleavable PEG which masks the TAT moieties, PTX-loaded liposomes have a long circulation time for enhanced tumor accumulation under normal physiologic condition. In the presence of GSH, however, PEG was removed and TAT was exposed, triggering a fast cellular uptake. The endocytosis inhibition study revealed that the active endocytosis of C-TAT-LP was mediated by

ACKNOWLEDGMENTS

This research was supported by the National Natural Science Foundation of China (81373337) and the National Basic Research Program of China (973 Program, 2013CB932504).

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