Design, synthesis and in vitro evaluation ofd-glucose-based cationic glycolipids for gene delivery

Abstract

Glycolipids might become a new type of promising non-viral gene delivery systems because of their low cytotoxicity, structural diversity, controllable aqua-and lipo-solubility, appropriate density and distribution of positive charges, high transfer efficiency and potential targeting function. In this study, four kinds of L-arabinose-based cationic glycolipids (Ara-DiC12MA, Ara-DiC14MA, Ara-DiC16MA and Ara-DiC18MA) containing quaternary ammonium as hydrophilic headgroup and two alkane chains as hydrophobic domain were synthesized and characterized. They were observed to have strong affinities for plasmid DNA (pDNA) and siRNA, the pDNA can be completely condensed at N/P ratio less than 2, and the siRNA can be completely retarded at N/P ratio less than 3. The dynamic light scattering (DLS) experiment and atomic force microscopy (AFM) experiment demonstrated that cationic lipids and their lipoplexes possessed suitable particle sizes with near-spherical shape and proper ζ-potentials for cell transfection. The Ara-DiC16MA liposome was found to have good trans-fection efficacy in HEK293, PC-3 and Mat cells compared with other three kinds of lipo-somes, and also maintain low cytotoxicity and better uptake capability in vitro. Furthermore, the gene silencing assay showed that Ara-DiC14MA and Ara-DiC16MA liposomes have demonstrated effective delivery and higher gene knockdown activity (>80%) in the above mentioned cells than Lipofectamine 2000. These results indicated Ara-DiC16MA can be developed for efficient and low toxic gene delivery.

Evaluation of Maltose-Based Cationic Liposomes with Different Hydrophobic Tails for Plasmid DNA Delivery

“…For Malt-DiC16MA liposome, the density of GFP cells increased with increase of N/P ratio but the fluorescent protein expression was weaker integrally than Malt-DiC14MA lipoplexes. However, the transfection efficiency of glucose-based lipids with the hydrophobic tails (C14 or C16) on the positively charged nitrogen atoms were as good as or better than lipo2000 [ 37 ]. Simultaneously, the lack of transfections of Malt-DiC12MA liposomes under different N/P ratios and the other two liposomes (Malt-DiC14MA and Malt-DiC16MA) at N/P of 2:1 were also observed, which corresponded with the glucose-based cationic liposomes with hydrophobic tails (C12) at all the N/P ratios and (C14 and C16) at low N/P ratios.…”

“…Among these cationic lipids, carbohydrates-based cationic lipids have been drawn great attention [ 28 , 29 , 30 , 31 ] because carbohydrates and their derivatives (such as, glycans, proteoglycans, glycoproteins, or glycolipids [ 32 , 33 ]) played key roles in a wide range of biological processes including cell-cell informational reception and transmission, cell growth and metastasis, cell adhesion intercellular identification and immunity [ 34 , 35 , 36 ]. Furthermore, lots of studies have shown that glycoside ligand could be acted as a targetable ligand for receptors, such as galactose, mannose, glucose and hyaluronic acid [ 30 , 31 , 37 , 38 , 39 ], which overexpressed in the surface of cancer cells. Therefore, carbohydrates may be conjugated in targeted gene delivery system with high transfer efficiency and safety.…”

“…In a previous study [ 37 ], we have demonstrated that glucose-based cationic lipids with hydrophobic tails linked to the positively charged nitrogen atom were higher than that linked to the sugar ring. To investigate the affection of sugar skeleton on the transfection efficiencies of sugar-based cationic lipids, in this work, three cationic lipids of Malt-DiC12MA ( IX a ), Malt -DiC14MA ( IX b ) and Malt-DiC16MA ( IX c ) ( Scheme 1 ), with hydrophobic chains linked to the positively charged nitrogen atoms, were successfully synthesized and the corresponding structures were confirmed by 1 H-NMR, 13 C-NMR, 1 H- 1 H COSY (2D-correlation spectroscopy) and 1 H- 13 C HSQC (Heteronuclear Single Quantum Coherence).…”

Synthesis and evaluation of L-arabinose-based cationic glycolipids as effective vectors for pDNA and siRNA in vitro

“…Guided by the structure-function relationship, lots of biodegradable and commercial available chemicals such as amino acids [41], oligopeptide [42], cholesterol [43, 44], cyclen [45] and carbohydrates [46, 47] were commonly selected as the backbone of cationic lipids to improve the transfection efficiency and targeting delivery. In recently, more and more carbohydrates-based cationic lipids [48], were designed and synthesized for the reason that carbohydrates are involved in many important physiological processes, including inflammatory and immunological responses, tumor metastasis, cell-cell signaling, apoptosis, cell adhesion, bacterial and viral recognition, and anticoagulation [49]. In addition, carbohydrates are specific ligands to targeting cell surface receptors, which receive significant attention in recent decades.…”

“…Receptor-mediated endocytosis is promoted to cellular uptake capacity in gene delivery [50]. The ligands corresponding to cell surface receptors including glucose [48, 51], galactose [50, 52], mannose[53], hyaluronic acid (HA) [54] and so on, which have been conjugated in gene delivery vectors.…”