ABSTRACT
A modular dendrimer-based drug delivery platform was designed to improve upon existing limitations in single dendrimer systems. Using this modular strategy, a biologically active platform containing receptor mediated targeting and fluorescence imaging modules was synthesized by coupling a folic acid (FA) conjugated dendrimer with a fluorescein isothiocyanate (FITC) conjugated dendrimer. The two different dendrimer modules were coupled via the 1,3-dipolar cycloaddition reaction ("click" chemistry) between an alkyne moiety on the surface of the first dendrimer and an azide moiety on the second dendrimer. Two simplified model systems were also synthesized to develop appropriate "click" reaction conditions and aid in spectroscopic assignments. Conjugates were characterized by (1)H NMR spectroscopy and NOESY. The FA-FITC modular platform was evaluated in vitro with a human epithelial cancer cell line (KB) and found to specifically target the overexpressed folic acid receptor.
Subject(s)
Dendrimers/metabolism , Drug Carriers/metabolism , Drug Delivery Systems , Drug Design , Folate Receptors, GPI-Anchored/analysis , Folic Acid/metabolism , Click Chemistry , Dendrimers/chemical synthesis , Dendrimers/chemistry , Drug Carriers/chemical synthesis , Drug Carriers/chemistry , Fluorescent Dyes/chemistry , Folate Receptors, GPI-Anchored/biosynthesis , Folic Acid/chemistry , Humans , Isothiocyanates/chemistry , KB Cells , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Structure , Surface Properties , Tumor Cells, CulturedABSTRACT
It has long been recognized that cationic nanoparticles induce cell membrane permeability. Recently, it has been found that cationic nanoparticles induce the formation and/or growth of nanoscale holes in supported lipid bilayers. In this paper, we show that noncytotoxic concentrations of cationic nanoparticles induce 30-2000 pA currents in 293A (human embryonic kidney) and KB (human epidermoid carcinoma) cells, consistent with a nanoscale defect such as a single hole or group of holes in the cell membrane ranging from 1 to 350 nm(2) in total area. Other forms of nanoscale defects, including the nanoparticle porating agents adsorbing onto or intercalating into the lipid bilayer, are also consistent; although the size of the defect must increase to account for any reduction in ion conduction, as compared to a water channel. An individual defect forming event takes 1-100 ms, while membrane resealing may occur over tens of seconds. Patch-clamp data provide direct evidence for the formation of nanoscale defects in living cell membranes. The cationic polymer data are compared and contrasted with patch-clamp data obtained for an amphiphilic phenylene ethynylene antimicrobial oligomer (AMO-3), a small molecule that is proposed to make well-defined 3.4 nm holes in lipid bilayers. Here, we observe data that are consistent with AMO-3 making approximately 3 nm holes in living cell membranes.
Subject(s)
Cations , Cell Membrane/metabolism , Nanoparticles , Cell Line , Humans , NanotechnologyABSTRACT
Both the R and S enantiomers of the amphibian alkaloid noranabasamine were prepared in >30% overall yield with 80% ee and 86% ee, respectively. An enantioselective iridium-catalyzed N-heterocyclization reaction with either (R)- or (S)-1-phenylethylamine and 1-(5-methoxypyridin-3-yl)-1,5-pentanediol was employed to generate the 2-(pyridin-3-yl)-piperidine ring system in 69-72% yield.
Subject(s)
Alkaloids/chemical synthesis , Iridium/chemistry , Piperidines/chemical synthesis , Pyridines/chemical synthesis , Alkaloids/chemistry , Amphibians , Animals , Catalysis , Molecular Structure , Piperidines/chemistry , Pyridines/chemistry , StereoisomerismABSTRACT
The interaction of generation 5 (G5) and 7 (G7) poly(amidoamine) (PAMAM) dendrimers with mica-supported Survanta bilayers is studied with atomic force microscopy (AFM). In these experiments, Survanta forms distinct gel and fluid domains with differing lipid composition. Nanoscale defects are induced by the PAMAM dendrimers. The positively charged dendrimers remove lipid from the fluid domains at a significantly greater rate than for the gel domains. Dendrimer accumulation on lipid edges and terraces preceding lipid removal has been directly imaged. Immediately following lipid removal, the mica surface is clean, indicating that lipid defects are not induced by dendrimers binding to the mica substrate and displacing the lipid.
