Self-assembled carbohydrate-based vesicles for lectin targeting
Graphical abstract
Introduction
The surface functionalization of nanoparticles and liposomes has become of increasing importance since Ehrlich publicized his vision of the “Magic Bullet” [1]. Vesicles and liposomes (phospholipid-based vesicles) have been extensively studied due to their widespread application as controlled drug delivery vehicles in the pharmaceutical industry and as biomembranes [2], [3], [4]. Because of their lipid composition, vesicles have lower toxicity than other vehicles, making them promising systems for the delivery of a wide range of drugs requiring specific treatments, controlled circulation times, reduced side effects and optimum drug action [5], [6], [7], [8], [9], [10], [11]. The possibility of directing a drug toward targeted tissues without changing its structure and hence, its biological activity, is fundamental for therapeutic applications [12]. Thus, efforts have been directed toward improving the vectorization of drug delivery systems to specific target tissues. In this context, the surface functionalization of vesicles with natural or synthetic glycolipids has been proposed to enhance the specificity of vesicles for lectins [12], [13]. Lectins are non-immunological proteins or glycoproteins that specifically recognize sugar molecules and are capable of binding glycosylated membrane components. They are widely used to characterize carbohydrates on cell surfaces [14]. The Bauhinia variegata lectin (BVL) is particularly found in Caesalpinoideae plants. Its subunit has molecular weight of ∼33 kDa and diffraction patter similar to related lectins such as Bauhinia purpurea agglutinin (BPA) [15]. The carbohydrate-lectin binding typically involves two or three terminal sugar residues of mammalian glycans, including galactose, mannose, N-acetyl-neuraminic acid, fucose, and N-acetyl-glucosamine [16], [17].
High levels of lectins, such as galectin-3, have been detected in various cancers [12]. The specificity of the carbohydrate-lectin interaction has been exploited to convey glycosylated liposomes to tumor cells [12], [18], [19], [20]. Therefore, the development of nanoparticles with outer shells decorated with glycoconjugates for lectin targeting is considered a promising means to improve the delivery and internalization of antitumor drugs [16], [21]. Previously, our research group described the physicochemical interactions between vesicles composed of phosphatidylcholine-purified soybean lecithin and the glycosylated polymeric amphiphile N-acetyl-β-d-glucosaminyl-PEG900-docosanate conjugate (C22PEG900GlcNAc). Structurally, the ∼100 nm composite vesicles self-assemble via attractive force between the lipidic region and the nitrogen groups of C22PEG900GlcNAc. The results also suggested discrete interaction among the hydroxyl and carbonyl regions of C22PEG900GlcNAc [22]. Amphiphiles containing PEG chains have become a topic of interest due to their biocompatibility and anomalous behavior in water [23]. Through the PEGylation of vesicles, nanoparticles, and proteins, the residence times of these carriers can be significantly extended and diminish their uptake by the organs (e.g., liver and spleen) of the reticulo endothelial system (RES) [24], [25].
This present work reports on the interaction between self-assembled vesicles (composed of phosphatidylcholine-purified soybean lecithin and the glycosylated polymeric amphiphile, C22PEG900GlcNAc) and BVL. The physicochemical properties of the phosphatidylcholine-based vesicle system containing C22PEG900GlcNAc and lectin were investigated by several techniques, including zeta potential, dynamic light scattering (DLS), atomic force microscopy (AFM), small-angle X-ray scattering (SAXS), horizontal attenuated total reflectance Fourier transform infrared (HATR-FTIR), differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis.) spectroscopy measurements. The physicochemical properties of these self-assembled vesicles presented herein could contribute to improved vectorizations of drug delivery systems in cancer therapy.
Section snippets
Materials
Phosphatidylcholine (PC) from soybean lecithin (95% phosphatidylcholine, 5% lysophosphatidylcholine and phosphatidic acid) was a gift from Solae do Brasil S.A. The molecular composition of the soybean PC was approximately 75% distearoylphosphatidylcholine (DSPC, 18:0), 12% dioleoylphosphatidylcholine (DOPC, 18:2) and 8% dipalmitoylphosphatidylcholine (DPPC, 16:0) [26]. All reagents were of commercial grade and were used as received unless otherwise noted. The native Bauhinia variegata (BVL)
Lectin effect on vesicle charge and size
The zeta potential (ZP) was used to evaluate the surface charge of the PC + C22PEG900GlcNAc vesicles in the presence of lectin. The zeta potential of PC + C22PEG900GlcNAc vesicles in phosphate buffer saline was −47 ± 2 mV, indicating stable assemblies in suspension due to electrostatic repulsion between particles, resulting in relative colloidal stability, as previously reported by the authors [22]. Fig. 1 shows the zeta potential values with increasing amounts of lectin. As the lectin concentration
Conclusion
We investigated the interaction between vesicles formed by phosphatidylcholine (PC) and the glycosylated polymeric amphiphile N-acetyl-β-d-glucosaminyl-PEG900-docosanate conjugate (C22PEG900GlcNAc) with the lectin BVL. Our results suggested that the lectin molecules recognized the glycosylated surfaces of the vesicles. Furthermore, the lectin molecules were located between the lipid choline and carbonyl groups, thereby changing the choline orientation and ordering the lipid polar head and
Conflicts of interest
The authors declare no conflicts of interest.
Acknowledgments
The authors wish to thank the Conselho Nacional de Pesquisa e Desenvolvimento (CNPq– Brazil) (440619/2014-9) for financial support. The LNLS is acknowledged for supplying the SAXS beam time (proposals 20150020 and 20150021). A.G.D.B acknowledges the financial support from FAPESC 3805/2012, and F.C.G. thanks FAPESP (grant 2014/22983-9).
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