New and more efficient multivalent glyco-ligands for asialoglycoprotein receptor of mammalian hepatocytes.
ABSTRACT New multi-valent, carbohydrate ligands that contain terminal N-acetylgalactosamine (GalNAc) or lactose (Lac) were prepared using a nitrilotriacetic acid (NTA) derivative of L-lysine as scaffold. Tri-valent structures were prepared by attaching an ω-amino glycoside of GalNAc or Lac to each of the three carboxyl groups of N(ε)-protected N(α)-dicarboxymethyl-L-lysine. In addition, a hexa-valent lactoside was synthesized by attaching N(ε)-deprotected trivalent lactoside to each of the carboxyl group of N(α)-(trifluoroacetamido)hexanoyl L-aspartic acid. Tri-valent GalNAc glycosides and the hexa-valent lactoside had high affinity (dissociation constants approaching nM) for rat hepatocytes. The hexa-valent lactoside, after de-N(ε)-protection, was modified with a chelator, diethylenetriaminepentaacetic acid (DTPA), through which a fluorescent or radioactive tag, such as europium or indium, can be firmly attached. Intravenous infusion of (111)Indium-tagged hexa-valent lactoside to rats and mice resulted in nearly exclusive accumulation of radioactivity in the liver.
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ABSTRACT: C-type lectin receptors (CLRs) represent a large receptor family including collectins, selectins, lymphocyte lectins, and proteoglycans. CLRs share a structurally homologous carbohydrate-recognition domain (CRD) and often bind carbohydrates in a Ca(2+)-dependent manner. In innate immunity, CLRs serve as pattern recognition receptors (PRRs) and bind to glycan structures of pathogens but also self-antigens. In nature, the low affinity of CLR/carbohydrate interactions is overcome by multivalent ligand presentation at the surface of cells or pathogens. Thus, multivalency is a promising strategy for targeting CLR-expressing cells and, indeed, carbohydrate-based targeting approaches have been employed for a number of CLRs, including asialoglycoprotein receptor (ASGPR) in the liver, or DC-SIGN expressed by dendritic cells. Since CLR engagement mediates endocytosis but also influences intracellular signaling pathways, CLR targeting may allow for cell-specific drug delivery and also the modulation of cellular functions. Glyconanoparticles, glycodendrimers, and glycoliposomes were successfully used as tools for CLR-specific targeting. This review will discuss different approaches for multivalent CLR ligand presentation and aims to highlight how CLR targeting has been employed for cell specific drug delivery. Major emphasis is directed towards targeting of CLRs expressed by antigen-presenting cells to modulate immune responses.Advanced drug delivery reviews 05/2013; · 11.96 Impact Factor
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ABSTRACT: We have synthesized poly-γ-glutamic acid (PGA) modified with a synthetic trivalent glyco-ligand (TriGalNAc) for the hepatocyte asialoglycoprotein receptor (ASGP-R). We investigated in vivo distribution of unmodified PGA and TriGalNAc-modified PGA (TriGalNAc-PGA) in mice after intravenous injection. Most of unmodified PGA administered was transported to the bladder over 20-80min, suggesting a rapid excretion of unmodified PGA into urine. In contrast, TriGalNAc-PGA was found exclusively in the liver over the same period of time. We further synthesized TriGalNAc-PGA-primaquine conjugate (TriGalNAc-PGA-PQ), and investigated binding, uptake, and catabolism of the conjugate by rat hepatocytes. Our studies indicated that approximately 250ng per million cells of the conjugate bound to one million rat hepatocytes at 0°C, and approximately 2μg per million cells of the conjugate was taken up over 7h incubation at 37°C. Furthermore, our results suggested that TriGalNAc-PGA-PQ was almost completely degraded over 24h, and small degradation products were secreted into cell culture medium. The results described in this report suggest that the TriGalNAc ligand can serve as an excellent targeting device for delivery of PGA-conjugates to the liver hepatocytes, and rat hepatocytes possess sufficient capacity to digest PGA even modified with other substituents.Bioorganic & medicinal chemistry 06/2013; · 2.82 Impact Factor
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ABSTRACT: Gelatin is one of the most versatile natural biopolymers widely used in pharmaceutical industries due to its biocompatibility, biodegradability, low cost and numerous available active groups for attaching targeting molecules. These advantages led to its application in the synthesis of nanoparticles for drug and gene delivery during the last thirty years. The current article entails a general review of the different preparation techniques of gelatin nanoparticles (GNPs): desolvation, coacervation-phase separation, emulsification-solvent evaporation, reverse phase microemulsion, nanoprecipitation, self-assembly and layer-by-layer coating, from the point of view of the methodological and mechanistic aspects involved. Various crosslinkers used to improve the physicochemical properties of GNPs including aldehydes, genipin, carbodiimide/N-hydroxysuccinimide, and transglutaminase are reported. An analysis is given of the physicochemical behavior of GNPs including drug loading, release, particle size, zeta-potential, cytotoxicity, cellular uptake and stability. This review also attempts to provide an overview of the major applications of GNPs in drug delivery and gene therapy and their in vivo pharmacological performances, as well as site-specific drug targeting using various ligands modifying the surface of GNPs. Finally, nanocomplexes of gelatin with polymers, lipids or inorganic materials are also discussed.Journal of Controlled Release 10/2013; · 7.63 Impact Factor