Water-soluble nanoparticles from random copolymer and oppositely charged surfactant, 3a. Nanoparticles of poly(ethylene glycol)-based cationic random copolymer and fatty acid salts

In this report, we investigate the nanoparticle formation between random copolymers (RCPs) of methoxy-poly(ethylene glycol) monomethacrylate (MePEGMA) and (3-(methacryloylamino)propyl)trimethylammonium chloride (MAPTAC) and oppositely charged natural surfactants, sodium oleate and sodium laurate, using turbidimetric titration, steady-state fluorescence, dynamic light scattering, and electron microscopy. Though sodium oleate and sodium laurate are sparingly soluble in water, the nanoparticle complexes formed between the RCPs and these surfactants are soluble in the entire range of compositions studied here, including the stoichiometric electronetural complexes. The spherical nature of these nanoparticle complexes is revealed by electron microscopic (EM) analysis. Dynamic light scattering (DLS) showed that the average diameters of the nanoparticles are in the range 50 to 150 nm, which is supported by EM analysis. Pyrene fluorescence experiments suggested that these soluble nanoparticles have hydrophobic cores, which may solubilize hydrophobic drug molecules. The polarity index (I(1)/I(3)) obtained from the pyrene fluorescence spectra and the conductometric measurements showed that the critical concentration of fatty acid salts needed to obtain nanoparticles are in the order of 10(-4) M. Further, the complexation of such poorly water-soluble amphiphilic surfactants with polymers offers a useful method for the immobilization of hydrophobic compounds towards water-soluble drug carrier formulations. The formation of water-soluble nanoparticles by the self-assembly of fatty acid salts upon interacting with oppositely charged poly(ethylene glycol)-based polyions.     

Water-soluble complexes from random copolymer and oppositely charged surfactant. 2. Complexes of poly(ethylene glycol)-based cationic random copolymer and bile salts

The complexes formed between the positively charged random copolymers (RCPs) of methoxy-poly(ethylene glycol) monomethacrylate (MePEGMA) and (3-(methacryloylamino)propyl)trimethylammonium chloride (MAPTAC) with oppositely charged biosurfactants (bile salts) were studied using turbidimetric titration, steady-state fluorescence, dynamic light scattering, and electron microscopy. Studies showed that the complexes of the RCPs of MAPTAC and MePEGMA with less than 68 mol % of PEG content precipitate in water, whereas the complexes of the copolymer with 89 and 94 mol % of PEG content do not precipitate in the entire range of composition of the mixture including stoichiometric compositions when the electroneutral complexes are formed. The complexes with true hydrophobic domains, which are a prerequisite characteristic to serve as a carrier, can be obtained at much lower concentration than the critical micelle concentration of the corresponding surfactant. For a particular surfactant, hydrophobic domains are obtained at lower Z-/+ for the random copolymer with lower PEG content. The hydrodynamic radii of these complexes vary over a range of 20-35 nm. Overall results reveal that these complexes are qualitatively similar to the polyion complex micelles or block ionomer complexes obtained from the block copolymers and oppositely charged surfactants. As the surfactants used in this study are biocompatible, we hope that these soluble particles will be promising vectors in the field of drug delivery.     

Improved complementary polymer pair system: switching for enzyme activity by PEGylated polymers

The development of technology for on/off switching of enzyme activity is expected to expand the applications of enzyme in a wide range of research fields. We have previously developed a complementary polymer pair system (CPPS) that enables the activity of several enzymes to be controlled by a pair of oppositely charged polymers. However, it failed to control the activity of large and unstable α-amylase because the aggregation of the complex between anionic α-amylase and cationic poly(allylamine) (PAA) induced irreversible denaturation of the enzyme. To address this issue, we herein designed and synthesized a cationic copolymer with a poly(ethylene glycol) backbone, poly(N,N-diethylaminoethyl methacrylate)-block-poly(ethylene glycol) (PEAMA-b-PEG). In contrast to PAA, α-amylase and β-galactosidase were inactivated by PEAMA-b-PEG with the formation of soluble complexes. The enzyme/PEAMA-b-PEG complexes were then successfully recovered from the complex by the addition of anionic poly(acrylic acid) (PAAc). Thus, dispersion of the complex by PEG segment in PEAMA-b-PEG clearly plays a crucial role for regulating the activities of these enzymes, suggesting that PEGylated charged polymer is a new candidate for CPPS for large and unstable enzymes.     

Dendronised block copolymers as potential vectors for gene transfection

A series of block copolymers containing a dendronised cationic block for efficient DNA binding and a poly(ethylene glycol) block for encapsulation of the complex were synthesised in a modular fashion using a combination of click chemistry and ring-opening metathesis polymerisation. DNA binding experiments, investigated using gel electrophoresis, dynamic light scattering and transmission electron microscopy, showed that all polymers prepared in this study strongly complex DNA and self-assemble into polyion complex micelles with apparent hydrodynamic radii ranging from 20-120 nm at physiological pH (7.4). The in vitro transfection efficiency and toxicity of these potential non-viral vectors were also evaluated in HeLadouble dagger cells using plasmid DNA encoding for green fluorescent protein as the reporter gene.     

Novel biodegradable polymers as gene carriers

This study investigated two new biodegradable polymers as gene controlled-released coatings for gene transfer. Poly(ethylene glycol)-co-poly(D,L-lactic acid) (PELA) and poly(ethylene glycol)-co-poly(lactic acid)-co-poly(glycolic acid) random copolymer (PELGA) were synthesized and used as microspheres matrices with encapsulated plasmid pCH110. The plasmid loading efficiency, cytotoxicity, transfection efficiency and in vitro degradation and release profiles of microsphere complexes were evaluated in details. The biodegradable polymers showed high DNA loading efficiency and low cytotoxicity as gene controlled-released coatings, and the poly(ethylene glycol) (PEG) contents of polymer matrices influenced the diameter, loading efficiency and transfection efficiency of plasmid DNA within the microspheres. The average diameters of PELA and PELGA microspheres were between 0.5 and 1.5 microm, and the plasmid loading efficiency was 62 and 73% for PELA and PELGA microspheres with 10% PEG content, respectively. In vitro testing showed a gradual release profile of DNA from polymeric matrices. The polymers/DNA microspheres had high transfection efficiency and early gene expression and maintenance of gene expression level for up to 96 h, although transfection efficiency were slightly lower than that of liposome in the initial 24 h. The biodegradable polymeric materials possess potential superiority as gene carriers.     

Enhanced dispersion stability of supramolecular complexes of single-walled carbon nanotubes with fluorene-based conjugated polymers

Supramolecular complexes of single-walled carbon nanotubes (SWNTs) with poly(9,9-didodecylfluorene-2,7-diyl) (PF) derivatives were prepared using a solution dispersion process. A series of novel conjugated PF polymers with carboxyl or hydroxyl end groups at both ends were synthesized by the Yamamoto-type coupling of 2,7-dibromo-9,9-didodecylfluorene using Ni(COD)₂ as a catalyst, and further end-capped with either 4-bromobenzoic acid or 4-bromobenzyl alcohol to obtain the end-functionalized PF with different terminal groups. An α-monocarboxy-ω-mono-methoxy poly(ethylene glycol) was connected to both ends of the PF-containing hydroxyl end groups to produce triblock copolymers of poly(ethylene glycol)-b-polyfluorene-b-poly(ethylene glycol) (PEO-b-PF-b-PEO). These SWNTs were completely wrapped with the conjugated polymers through π–π interactions, which enhanced the solubility of the SWNT complexes in organic media, and prevented the aggregation of the polymer–SWNT complexes into large bundles. This indicates that the dispersion stability of SWNTs is enhanced by the addition of the conjugated polymers.     

Novel Biodegradable Block Copolymers of Poly(ethylene glycol) (PEG) and Cationic Polycarbonate: Effects of PEG Configuration on Gene Delivery

A novel amine‐functionalized polycarbonate was synthesized and its excellent gene transfection ability in vitro is demonstrated. In the framework of adapting the cationic polycarbonate for in vivo gene delivery applications, here the design and synthesis of biodegradable block copolymers of poly(ethylene glycol) (PEG) and amine‐functionalized polycarbonate with a well‐defined molecular architecture and molecular weight is achieved by metal‐free organocatalytic ring‐opening polymerization. Copolymers in triblock cationic polycarbonate‐block‐PEG‐block‐cationic polycarbonate and diblock PEG‐block‐cationic polycarbonate configurations, in comparison with a non‐PEGylated cationic polycarbonate control, are investigated for their influence on key aspects of gene delivery. Among the polymers with similar molecular weights and N content, the triblock copolymer exhibit more favorable physicochemical (i.e., DNA binding, size, zeta‐potential, and in vitro stability) and biological (i.e., cellular uptake and luciferase reporter gene expression) properties. Importantly, the various cationic polycarbonate/DNA complexes are biocompatible, inducing minimal cytotoxicities and hemolysis. These results suggest that the triblock copolymer is a more useful architecture in future cationic polymer designs for successful systemic therapeutic applications.     

Inclusion complex formation between α,γ‐cyclodextrins and organic–inorganic star‐shaped poly(ethylene glycol) from an octafunctional silsesquioxane core

A series of organic–inorganic star‐shaped poly(ethylene glycol)s from octafunctional silsesquioxane (POSS) cores were synthesized by allylation and hydrosilylation, and they were found to form crystalline inclusion complexes with α,γ‐cyclodextrins. The results from X‐ray diffraction, cross‐polarity/magic‐angle‐spinning ¹³C NMR, differential scanning calorimetry, and Fourier transform infrared suggested that a channel‐type structure was established in the inclusion complexes. The characterization results also revealed that the segments of poly(ethylene glycol) arms near the POSS cores were uncovered by the cyclodextrins, whereas the cyclodextrins were closely packed along the remaining portion of the poly(ethylene glycol) arms. Molecular dynamic simulation in a Dreiding force field with Cerius 2 software gave comparable results. © 2004 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 42: 1173–1180, 2004     

Microporous structure and drug release kinetics of polymeric nanoparticles

The aim of the present study was to characterize pegylated nanoparticles (NPs) for their microporosity and study the effect of microporosity on drug release kinetics. Blank and drug-loaded NPs were prepared from three different pegylated polymers, namely, poly(ethylene glycol)1%-graft-poly(D,L)-lactide, poly(ethylene glycol)5%-graft-poly(D,L)-lactide, and the multiblock copolymer (poly(D,L)-lactide-block-poly(ethylene glycol)-block-poly(D,L)-lactide)n. These NPs were characterized for their microporosity using nitrogen adsorption isotherms. NPs of the multiblock copolymer showed the least microporosity and Brunauer-Emmett-Teller (BET) surface area, and that of PEG1%-g-PLA showed the maximum. Based on these results, the structural organization of poly(D,L)-lactide (PLA) and poly(ethylene glycol) (PEG) chains inside the NPs was proposed and was validated with differential scanning calorimetry (DSC) and X-ray photoelectron spectroscopy (XPS) surface analysis. An in vitro drug release study revealed that PEG1%-g-PLA NPs exhibited slower release despite their higher surface area and microporosity. This was attributed to the presence of increased microporosity forming tortuous internal structures, thereby hindering drug diffusion from the matrix. Thus, it was concluded that the microporous structure of NPs, which is affected by the molecular architecture of polymers, determines the release rate of the encapsulated drug.     

Specific features of nonstoichiometric interpolymer complexes of poly(acrylic acid) and poly(ethylene glycol) as protectors of copper nanoparticles in aqueous sols

Advantages of interpolymer complexes for use as amphiphilic protectors of nanoparticles during the formation and stabilization of sols are considered. The effects of the ratio of poly(acrylic acid) and poly(ethylene glycol) and the molecular mass of poly(ethylene glycol) on the mean size and size distribution of copper nanoparticles in sols formed via the reduction of divalent copper ions in mixed aqueous solutions of these polymers are investigated. It is shown that sols of metal nanoparticles with small sizes and narrow size distributions are formed even when poly(ethylene glycols) with chain lengths below the “critical” chain length and a small PEG-to-PAA base-molar ratio are used. This is evidence for efficient protection of the formed copper nanoparticles by the interpolymer complex PEG-PAA under conditions of its instability and for self-organization of oligomeric PEG chains in complex macromolecular shields of nanoparticles.     

Thermogelling polymers composed of poly(cyclohexylenedimethylene adipate) and poly(ethylene glycol)

Block copolymers composed of hydrophilic poly(ethylene glycol) (PEG) and hydrophobic biodegradable polyesters have been reported as thermogelling polymers, because they feature temperature-dependent sol-to-gel or gel-to-sol transitions in aqueous solutions. In this study, a series of thermogelling poly(ethylene glycol methyl ether)-block–poly(cyclohexylenedimethylene adipate)-block–poly(ethylene glycol methyl ether) triblock copolymers and PEG-block–poly(cyclohexylenedimethylene adipate) multiblock copolymers was synthesized by reacting hydroxyl-terminated poly(cyclohexylenedimethylene adipate) (PCA) with poly(ethylene glycol methyl ether) and PEG, respectively, using 1,6-diisocyanatohexane as the coupling agent. Two hydroxyl-terminated PCAs, i.e., poly(1,4-cyclohexylenedimethylene adipate) and poly(1,3/1,4-cyclohexylenedimethylene adipate), were synthesized by the condensation reaction of adipic acid (AA) with 1,4-cyclohexanedimethanol (CHDM) and 1,3/1,4-CHDM, respectively, and used as the hydrophobic polyester blocks of these thermogelling copolymers to compare the effect of crystallinity on the sol-to-gel transition behavior.The polymers were characterized using proton nuclear magnetic resonance, Fourier transform infrared spectroscopy, gel permeation chromatography, differential scanning calorimetry, solubility testing, and rheological analysis. Experimental results revealed that the structure of the PCA block (crystalline vs. amorphous), the molecular weights of the hydrophobic PCA and hydrophilic PEG blocks, and the type of thermogelling polymer (triblock vs. multiblock) influenced the solubility, polymer micelle packing characteristics, maximum storage modulus, and sol-to-gel temperature of the polymers. Among all the samples at 40 wt.% aqueous solutions, triblock copolymer TB3 showed sol-to-gel temperature at 22 °C, and had the highest maximum storage modulus about 170 Pa.     

Solvent-free synthesis and purification of poly[2-(dimethylamino)ethyl methacrylate] by atom transfer radical polymerization

Solvent-free synthesis of well-defined poly[2-(dimethylamino)ethyl methacrylate] (PDMAEMA) (co)polymers was performed by atom transfer radical polymerization conducted under very mild conditions (in bulk at 25 degrees C). The pH-dependence and the thermo-responsive behaviour of PDMAEMA in aqueous solution were operated to isolate and purify the (co)polymers without using any organic solvent or further catalyst extraction. The viscosity in aqueous solution of so-purified PDMAEMA homopolymers and their block copolymers with poly(ethylene glycol) (PEG) was studied as a function of molar mass and concentration and a typical polyelectrolyte behaviour was observed, these catalyst-deprived polycations are able to form stable and non toxic complexes with DNA, showing good transfection efficacies in gene therapy.     

Decorated rods: a "bottom-up" self-assembly of monomolecular DNA complexes

Fluorescence correlation spectroscopy (FCS) and gel electrophoresis measurements are performed to investigate both the number and size of complexes of linear double-stranded DNA (dsDNA) fragments with 1:1 diblock copolymers consisting of a cationic moiety, branched polyethyleneimine (bPEI) of 2, 10, or 25 kDa, covalently bound to a neutral shielding moiety, poly(ethylene glycol) (PEG; 20 kDa). By systematically decreasing the bPEI length, the PEG grafting density along the DNA chain can be directly controlled. For 25 and 10 kDa bPEI-PEG copolymers, severe aggregation is observed despite the presence of the shielding PEG. Upon decreasing the bPEI length to 2 kDa, controlled self-assembly of monomolecular DNA nanoparticles is observed. The resulting complexes are in quantitative agreement with a theoretical model based on a single DNA encased in a dense PEG polymer brush layer. The resulting PEGylated complexes show high stability against both salt and protein and hence are of potential use for in vivo gene delivery studies.     

Threading, growth, and aggregation of pseudopolyrotaxanes

The formation of supramolecular inclusion compounds (pseudopolyrotaxanes) produced by cyclodextrins and polymers can be monitored through turbidimetry. In this paper, we studied the kinetics of the threading of linear poly(ethylene glycol) chains of different molecular weights and of a four-arm star-like polymer as a function of temperature. The main thermodynamic parameters are extracted. The aggregation and precipitation of pseudopolyrotaxanes are described in terms of the Avrami-Erofe'ev model, which provides relevant information on the mechanism of these processes. SAXS and TGA experiments confirm the structure and hydration of the final products obtained from the different polymers. A new hypothesis for the interaction between pseudopolyrotaxanes that leads to aggregation and precipitation, based on the spatial dielectric anisotropy, is proposed.     

Thermodynamic Study of Poly(ethylene glycol) in Water/1-Propanol Solutions by Viscometry

In this work, the intrinsic viscosities of poly(ethylene glycol) with a molar mass of 20 kg⋅mol⁻¹ were measured in water/1-propanol solutions from 283.1 to 313.1 K. The expansion factors of the polymer chains were calculated from the intrinsic viscosity data. The thermodynamic parameters entropy of dilution parameter, the heat of dilution parameter, theta temperature, polymer–solvent interaction parameter and second osmotic virial coefficient were derived from the temperature dependence of the polymer chain expansion factor. The thermodynamic parameters indicate that mixtures of water/1-propanol become weaker solvents for poly(ethylene glycol) with increasing temperature. Also, the thermodynamic parameters indicate that the solvent ability of mixed water/1-propanol for poly(ethylene glycol) is less than that of pure water.     

Desorption electrospray ionisation mass spectrometry and tandem mass spectrometry of low molecular weight synthetic polymers

A range of low molecular weight synthetic polymers has been characterised by means of desorption electrospray ionisation (DESI) combined with both mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Accurate mass experiments were used to aid the structural determination of some of the oligomeric materials. The polymers analysed were poly(ethylene glycol) (PEG), polypropylene glycol (PPG), poly(methyl methacrylate) (PMMA) and poly(alpha-methyl styrene). An application of the technique for characterisation of a polymer used as part of an active ingredient in a pharmaceutical tablet is described. The mass spectra and tandem mass spectra of all of the polymers were obtained in seconds, indicating the sensitivity of the technique.     

LASER FLASH SPECTROSCOPY OF METHYLENE BLUE WITH NUCLEIC ACIDS

Absorption spectra, fluorescence spectra and quantum yields, triplet state absorption spectra, yields and rates of decay have been obtained for methylene blue (MB+), MB+/CT-DNA complexes and complexes of MB+ with the synthetic polynucleotides poly[d(G-C)] and poly[d(A-T)]. Strong fluorescence quenching is observed in the complexes of the dye with CT-DNA and poly[d(G-C)] concomitant with a decrease in the triplet yield. The fluorescence and triplet yield of MB+ with poly[d(A-T)] are similar to those of the free dye. The triplet decay times are increased in all three polymer/dye complexes and show a decreased sensitivity to oxygen quenching. These results are interpreted in terms of the binding of the dye to these polymers and the implications of polymer binding on the photosensitizing properties of the dye are discussed.     

Poly(ethylene glycol)‐block‐Poly(glycidyl methacrylate) with Oligoamine Side Chains as Efficient Gene Vectors

Well‐defined diblock copolymers, poly(ethylene glycol)‐block‐poly(glycidyl methacrylate)s (PEG‐b‐PGMAs), with different poly(glycidyl methacrylate) (PGMA) chains, were prepared via atom transfer radical polymerization (ATRP) from the same macromolecular initiator 2‐bromoisobutyryl‐terminated poly(ethylene glycol) (PEG). Ethyldiamine (EDA), diethylenetriamine (DETA), triethylenetetramine (TETA), and polyethyleneimine (PEI) with an of 400 (PEI₄₀₀) were used to decorate PEG‐b‐PGMAs to get the cationic polymers PEG‐b‐PGMA‐ oligoamines. These cationic polymers possessed high buffer capability and could condense plasmid DNA (pDNA) into nanoscaled complexes of 125–530 nm. These complexes showed the positive zeta potential of 20–35 mV at N/P ratios of 10–50. Most of them exhibited very low cytotoxicity and good transfection efficiency in 293T cells. The presence of the serum medium did not decrease the transfection efficiency due to the steric stabilization of the PEG chains.

     

Investigation of DNA condensing properties of amphiphilic triblock cationic polymers by atomic force microscopy

Introduction of nucleic acids into cells is an important biotechnology research field which also holds great promise for therapeutic applications. One of the key steps in the gene delivery process is compaction of DNA into nanometric particles. The study of DNA condensing properties of three linear cationic triblock copolymers poly(ethylenimine-b-propylene glycol-b-ethylenimine), namely, LPEI(50)-PPG(36)-LPEI(50), LPEI(19)-PPG(36)-LPEI(19), and LPEI(14)-PPG(68)-LPEI(14), indicates that proper DNA condensation is driven by both the charge and the size of the respective cationic hydrophilic linear polyethylenimine (LPEI) and neutral hydrophobic poly(propylene glycol) (PPG) parts. Atomic force microscopy was used to investigate the interactions of the triblock copolymers with plasmid DNA at the single molecule level and to enlighten the mechanism involved in DNA condensation.     

Synthesis and Characterization of α,ω‐bi[2,4‐dinitrophenyl (DNP)] poly(2‐methoxystyrene) Functional Polymers. Initial Evaluation of the Interaction of the Functional Polymers with RBL Mast Cells

Two series of functional polymers, α,ω‐bi[2,4‐dinitrophenyl][poly(ethylene oxide)‐b‐poly(2‐methoxystyrene)‐b‐poly(ethylene oxide)] (DNP‐PEO‐P2MS‐PEO‐DNP) and α,ω‐bi[2,4‐dinitrophenyl caproic][poly(ethylene oxide)‐b‐poly(2‐methoxystyrene)‐b‐poly(ethylene oxide)] (CDNP‐PEO‐P2MS‐PEO‐CDNP), were synthesized by anionic living polymerization. The polymers were characterized by FT‐IR, ¹H‐NMR and Gel Permeation Chromatography (GPC). The molecular weight distributions for the lower molecular weight functional polymers were slightly broad (1.3–1.5). However, the molecular weight distributions for higher molecular weight polymers were narrower (1.1–1.2). Differential scanning calorimetry (DSC) studies showed thermal transitions indicative of the presence of microphases in the polymer solid state. The polymers were white powders and soluble in tetrahydrofuran. The binding affinity of DNP‐PEO‐P2MS‐PEO‐DNP ligands towards anti DNP IgE was determined by titrations with fluorescently labeled FITC‐IgE. A water soluble CDNP‐PEO‐P2MS‐PEO‐CDNP/DMEG (dimethoxyethylene glycol) complex binds and achieves steady state binding with solution IgE within a few seconds. This strongly suggests that CDNP functional polymers with improved water solubility have potential in therapeutics. Higher molecular weight (water insoluble) CDNP‐PEO‐P2MS‐PEO‐CDNP polymers were electrosprayed as fibers (500 nm) on silicon surface. Fluorescence spectroscopy clearly showed that RBL mast cells were interacting with the fibers suggesting that the cell‐surface receptors were clustered along the fiber surface. These observations suggest that the functional polymers hold promise for developing an antibody detection device.