Synthesis and characterization of amphiphilic polyphosphates with hydrophilic graft chains and cholesteryl groups as nanocarriers

Amphiphilic polyphosphate graft copolymers with varied densities of cholesteryl esters and hydrophilic graft chains were prepared, and the solution properties of the graft copolymers were evaluated. Polyphosphates were synthesized as backbones by ring-opening polymerization of 2-isopropyl-2-oxo-1,3,2-dioxaphospholane (IPP), 2-(2-oxo-1,3,2-dioxaphosphoroyloxyethyl-2-bromoisobutyrate) (OPBB), and 2-choresteryl-2-oxo-1,3,2-dioxaphospholane (ChOP) using triisobutylaluminum as an initiator. Three types of polyphosphates (PIBr(x)Ch(y), x = number of OPBB units in a polymer; y = number of ChOP units in a polymer) such as PIBr4, PIBr6Ch1, and PIBr3Ch2 were obtained. The molecular weights of these polymers were 2.4 x 10(4), 2.4 x 10(4), and 2.6 x 10(4) g/mol, respectively. 2-Methacryloyloxyethyl phosphorylcholine (MPC) was grafted from the OPBB sites in PIBr(x)Ch(y) via atom transfer radical polymerization (ATRP) in EtOH. In each polymer system, the molecular weight of the graft polymer was linear with conversion. Furthermore, the polymer radical concentration remained constant during polymerization; that is, the molecular weights of the graft chains were easily controllable with polymerization time. The solution properties of amphiphilic PIBr(x)Ch(y)-g-PMPCs were investigated by the methods of surface tension measurement, light scattering, and fluorescence probe. The transition point (cmc) of the surface tension of the PIBr(x)Ch(y)-g-PMPCs aqueous solution decreased with an increase in the number of ChOP units in a graft polymer. Particularly, PIBr3Ch2-g-PMPC14.9K formed nanosized associates (R(h) = 7.5 nm) with 2.2 molecules above 0.1 wt %. v79 cells were used to evaluate the cytotoxicity of the graft polymers, but no cytotoxicity was observed. The graft polymers containing cholesteryl groups effectively enhanced the solubility of paclitaxel in an aqueous solution.     

Synthesis and self-assembly of well-defined poly(amino acid) end-capped poly(ethylene glycol) and poly(2-methyl-2-oxazoline)

Poly(ethylene glycol) (PEG) and poly(2-methyl-2-oxazoline) (PMOx) are water-soluble, biocompatible polymers with stealth hemolytic activities. Poly(amino acid) (PAA) end-capped PEG and PMOx were prepared using amino-terminated derivatives of PEG and PMOx as macroinitiators for the ring-opening polymerization of γ-benzyl protected l-glutamate N-carboxyanhydride and S-benzyloxycarbonyl protected l-cysteine N-carboxyanhydride, respectively, in the presence of urea, at room temperature. The molecular weight of the PAA moiety was kept between M(n) = 2200 and 3000 g mol(-1). PMOx was polymerized by cationic ring-opening polymerization resulting in molecular weights of M(n) = 5000 and 10,000 g mol(-1), and PEG was a commercial product with M(n) = 5000 g mol(-1). Here, we investigate the self-assembly of the resulting amphiphilic block copolymers in water and the effect of the chemical structure of the block copolymers on the solution properties of self-assembled nanostructures. The PEG-block-poly(amino acid), PEG-b-PAA, and PMOx-block-poly(amino acid), PMOx-b-PAA, block copolymers have a narrow and monomodal molecular weight distribution (PDI < 1.3). Their self-assembly in water was studied by dynamic light scattering and fluorescence spectroscopy. In aqueous solution, the block copolymers associate into particles with hydrodynamic radii (R(H)) ranging in size from R(H) 70 to 130 nm, depending on the block copolymer architecture and the polymer molecular weight. Larger R(H) and critical association concentration values were obtained for copolymers containing poly(S-benzyloxycarbonyl-l-cysteine) compared to their poly(γ-benzyl-L-glutamate) analogue. FTIR investigations revealed that the poly(γ-benzyl-L-glutamate) block adopts a helical conformation, while the poly(S-benzyloxycarbonyl-L-cysteine) block exists as β-sheet.     

Low molecular weight linear polyethylenimine-b-poly(ethylene glycol)-b-polyethylenimine triblock copolymers: synthesis, characterization, and in vitro gene transfer properties

Novel ABA triblock copolymers consisting of low molecular weight linear polyethylenimine (PEI) as the A block and poly(ethylene glycol) (PEG) as the B block were prepared and evaluated as polymeric transfectant. The cationic polymerization of 2-methyl-2-oxazoline (MeOZO) using PEG-bis(tosylate) as a macroinitiator followed by acid hydrolysis afforded linear PEI-PEG-PEI triblock copolymers with controlled compositions. Two copolymers, PEI-PEG-PEI 2100-3400-2100 and 4000-3400-4000, were synthesized. Both copolymers were shown to interact with and condense plasmid DNA effectively to give polymer/DNA complexes (polyplexes) of small sizes (<100 nm) and moderate zeta-potentials (approximately +10 mV) at polymer/plasmid weight ratios > or =1.5/1. These polyplexes were able to efficiently transfect COS-7 cells and primary bovine endothelial cells (BAECs) in vitro. For example, PEI-PEG-PEI 4000-3400-4000 based polyplexes showed a transfection efficiency comparable to polyplexes of branched PEI 25000. The transfection activity of polyplexes of PEI-PEG-PEI 4000-3400-4000 in BAECs using luciferase as a reporter gene was 3-fold higher than that for linear PEI 25000/DNA formulations. Importantly, the presence of serum in the transfection medium had no inhibitive effect on the transfection activity of the PEI-PEG-PEI polyplexes. These PEI-PEG-PEI triblock copolymers displayed also an improved safety profile in comparison with high molecular weight PEIs, since the cytotoxicity of the polyplex formulations was very low under conditions where high transgene expression was found. Therefore, linear PEI-PEG-PEI triblock copolymers are an attractive novel class of nonviral gene delivery systems.     

Quartz crystal microbalance for the determination of daminozide using molecularly imprinted polymers as recognition element

As the daminozide (DM) and its metabolite have been identified to be potentially carcinogenic, rapid detection method for them is necessary for food safety. A type of piezoelectric crystal sensor has been prepared by using a molecularly imprinted polymer (MIP) as recognition element. The molecularly imprinted polymer was prepared by hot-induced precipitation polymerization, and then the polymer particles were fixed on the surface of the electrode. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were employed to evaluate the obtained imprinted polymer particles and the MIP sensitive film coated on the electrode. The results showed that a typical time-response curve of the MIP-coated crystal to the DM solution had been given, frequency shifts versus logarithm changes of DM showed good linear correlation within the concentration range of 1.0x10(-9) to 10(-6) mg/mL (y=11.38 lg x+115.45, r=0.9872) and 1.0x10(-6) to 10(-1) mg/mL (y=25.22lgx+209.44, r=0.9938), respectively. The detection limit was 5.0x10(-8) mg/mL (S/N=3), which is lower than that of conventional methods. Further, computer simulation technology was employed to investigate the interaction between methacrylic acid and DM for elucidating the recognition mechanism. The influencing factor pH has also been investigated. The injection experiments of DM structurally related compounds indicated that the obtained sensor has high sensitivity, excellent selectivity, low cost, good reproducibility, and reusable property by combining with piezoelectric crystal and molecularly imprinted polymer.     

Polyvalent side chain peptide-synthetic polymer conjugates as HIV-1 entry inhibitors

This report describes the synthesis and properties of a series of polyvalent side chain peptide-synthetic polymer conjugates designed to block the CD4 binding site on gp120 and inhibit HIV-1 entry into a host cell. The peptide sequences in the conjugates are based on the CDR H3 region of the neutralizing anti-HIV-1 antibody IgG1 b12. Using a consecutive ester-amide/thiol-ene postpolymerization modification strategy, a library of polymer conjugates was prepared. Evaluation of the HIV-1 inhibitory properties revealed that midsized polymer conjugates displayed the highest antiviral activity, while shorter and longer conjugates proved to be less efficacious inhibitors. The lower molecular weight conjugates may not have sufficient length to span the distance between two neighboring gp120 containing spikes, while the higher molecular weight conjugates may be compromised due to a higher entropic penalty that would accompany their binding to the viral envelope. Although the IC(50) values for these polymer conjugates are higher than that of the parent IgG1 b12 antibody, the strategy presented here may represent an interesting antiviral approach due to the attractive properties of such polymer therapeutics (relatively inexpensive production and purification costs, high thermal and chemical stability in storage conditions, long half-life in biological tissues, low immunogenicity, and protection from proteolytic degradation).     

Preparation of phenylalanine imprinted polymer by the sol–gel transition method

A phenylalanine (Phe) imprinted polymer was prepared by the wet-phase inversion and sol–gel transition method to endow a copolymer matrix with a large uptake capacity of template molecules and prominent adsorption selectivity at the high concentration of the racemate solution. A copolymer bead prepared by wet-phase inversion was shrunken in a hydrochloric acid solution containing a large amount of template molecules after swelling in a sodium hydroxide solution. Template molecules were effectively implanted in the polymer matrix during shrinking after swelling. The adsorption selectivities of Phe-imprinted copolymer bead were 2.1 and 1.33 at 1 g and 10 g Phe/l racemate solution, respectively, and the Phe uptake capacity reached about 1 g Phe/g dry weight of the copolymer. The adsorption selectivity of the copolymer was retained after five batches of adsorption/desorption in 1 g Phe/l solution composed of 5% D-Phe and 95% L-Phe.     

Highly efficient gene transfer with degradable poly(ester amine) based on poly(ethylene glycol) diacrylate and polyethylenimine in vitro and in vivo

BACKGROUND: Polyethylenimine (PEI) is toxic although it is one of the most successful and widely used gene delivery polymers with the aid of the proton sponge effect. Therefore, development of new novel gene delivery carriers having high efficiency with less toxicity is necessary. METHODS: In this study, a degradable poly(ester amine) carrier based on poly(ethylene glycol) diacrylate (PEGDA) and low molecular weight linear PEI was prepared. Furthermore, we compared the gene expression of the polymer/DNA complexes using two delivery methods: intravenous administration as an invasive method and aerosol as a non-invasive method. RESULTS: The synthesized polymer had a relatively small molecular weight (MW = 7980) with 25 h half-life in vitro. The polymer/DNA complexes were formed at an N/P ratio of 9. The particle sizes and zeta-potentials of the complexes were dependent on N/P ratio. Compared to PEI 25K, the newly synthesized polymer exhibited high transfection efficiency with low toxicity. Poly(ester amine)-mediated gene expression in the lung and liver was higher than that of the conventional PEI carrier. Interestingly, non-invasive aerosol delivery induced higher gene expression in all organs compared to intravenous method in an in vivo mice study. Such an expressed gene via a single aerosol administration in the lung and liver remained unchanged for 7 days. CONCLUSIONS: Our study demonstrates that poly(ester amine) may be applied as an useful gene carrier.     

Sugarcane bagasse derivative-based superabsorbent containing phosphate rock with water-fertilizer integration

To improve the water-fertilizer utilization ratio and mitigate the environmental contamination, an eco-friendly superabsorbent polymer (SPA), modified sugarcane bagasse/poly (acrylic acid) embedding phosphate rock (MSB/PAA/PHR), was prepared. Ammonia, phosphate rock (PHR) and KOH were admixed in the presence of acrylic acid to provide nitrogen (N), phosphorus (P) and potassium (K) nutrients, respectively. Impacts on water absorption capacity of the superabsorbent polymer (SAP) were investigated. The maximum swelling capacity in distilled water and 0.9wt.% (weight percent) NaCl solution reached 414gg(-1) and 55gg(-1) (water/prepared SAP), respectively. The available NPK contents of the combination system were 15.13mgg(-1), 6.93mgg(-1) and 52.05mgg(-1), respectively. Moreover, the release behaviors of NPK in the MSB/PAA/PHR were also studied. The results showed that the MSB/PAA/PHR has outstanding sustained-release plant nutrients property.     

Biodegradable molecularly imprinted polymers based on poly(ε-caprolactone)

Novel biodegradable molecularly imprinted polymers (MIPs) based on poly(ε-caprolactone) (PCL) were prepared by combining two important properties required of ideal biomaterials, biodegradability (with biocompatibility) and molecular recognition properties. Acrylate or methacrylate end-capped PCL macromers were synthesized through the reaction of PCL diol or triol with acryloyl or methacryloyl chloride. The synthesis of acrylate or methacrylate end-capped macromers was confirmed using FT-IR and H NMR spectroscopic techniques. These macromers were used to prepare biodegradable crosslinked networks by photopolymerization with functional monomer (acrylic acid) and a model template (theophylline). The theophylline-imprinted polymer showed higher binding capacity for theophylline compared with non-imprinted polymer (NIP), and also showed selectivity for theophylline over caffeine (similar structure molecules). PCL-based MIP degraded 8% of the initial weight in 30 days in phosphate-buffered saline (PBS) solution (pH 7.4) and over 90% of the initial weight within 24 h in 1 N NaOH at 37°C.     

Mucoadhesive vaginal tablets as veterinary delivery system for the controlled release of an antimicrobial drug,acriflavine

The aim of the study was the development of mucoadhesive vaginal tablets designed for the local controlled release of acriflavine, an antimicrobial drug used as a model. The tablets were prepared using drug-loaded chitosan microspheres and additional excipients (methylcellulose, sodium alginate, sodium carboxymethylcellulose, or Carbopol 974). The microspheres were prepared by a spray-drying method, using the drug to polymer weight ratios 1:1 and 1:2 and were characterized in terms of morphology, encapsulation efficiency, and in vitro release behavior, as MIC (Minimum Inhibitory Concentration), MBC (Minimum Bacterial Concentration), and killing time (KT). The tablets were prepared by direct compression, characterized by in vitro drug release and in vitro mucoadhesive tests. The microparticles have sizes of 4 to 12 microm; the mean encapsulation yields are about 90%. Acriflavine, encapsulated into the polymer, maintains its antibacterial activity; killing time of the encapsulated drug is similar to that of the free drug. In vitro release profiles of tablets show differences depending on the excipient used. In particular Carbopol 974, which is highly cross-linked, is able to determine a drug-controlled release from the matrix tablets for more than 8 hours. The in vitro adhesion tests, carried out on the same formulation, show a good adhesive behavior. The formulation containing microspheres with drug to polymer weight ratios of 1:1 and Carbopol 974 is characterized by the best release behavior and shows good mucoadhesive properties. These preliminary data indicate that this formulation can be proposed as a mucoadhesive vaginal delivery system for the controlled release of acriflavine.     

Novel Biodegradable Polyester Poly(Propylene Succinate): Synthesis and Application in the Preparation of Solid Dispersions and Nanoparticles of a Water-Soluble Drug

Poly(propylene succinate) (PPSu) polymers of average molecular weights from 2,800 to 13,100 g/mol were synthesized and characterized with regard to crystallinity, thermal properties, and cytocompatibility. Higher molecular weight samples exhibited lower degree of crystallinity and melted at lower temperatures. Melting of the polymer appeared to begin at 38°C. PPSu cytocompatibility was investigated based on human umbilical vein endothelial cells viability in the presence of increasing concentrations of polymer, and it was found that PPSu exhibited comparable cytocompatibility with poly(dl-lactide). The feasibility of applying PPSu as a drug carrier was shown for the first time, as solid dispersions and nanoparticles of sodium fluvastatin based in PPSu were prepared. Drug release rates decreased with increasing the molecular weight of PPSu in both solid dispersions and nanoparticles. For dispersions prepared from PPSu of the same molecular weight, drug release rates increased with drug loading. It appears that PPSu applicability as a drug carrier warrants further consideration.     

Heparin interferes with the inhibition of neutrophil elastase by its physiological inhibitors.

Heparin depresses the second-order rate constant kass for the inhibition of neutrophil elastase by alpha 1-proteinase inhibitor. For high and low molecular weight heparin the decrease in kass is 290-fold and 40-fold, respectively. This is due to a tight binding of the polymer to elastase: Kd = 3.3 nM or 89 nM for high or low molecular weight heparin respectively. In contrast heparin increases the rate of inhibition of elastase by mucus proteinase inhibitor. For low molecular weight heparin, there is a 27-fold increase in kass. This is due to a strong binding of the polymer to the inhibitor (Kd = 50 nM) which undergoes a conformational change.     

Texture properties of high and low acyl mixed gellan gels

The strength, deformability, and firmness of high acyl (H) and low acyl (L) mixed gellan gels were studied by compression tests. The gels were prepared with total polymer concentrations of 0.5, 1.0, and 1.5% at H/L weight ratios of 25/75, 50/50, 75/25, and calcium concentrations 2–80 mM. The mixed gels were much more deformable, with failure normal strains ranging from 0.6 to 1.5, but had similar strength compared to low acyl gellan gels. Both H/L ratio and total polymer concentration affected the textural properties, but H/L ratio was a more important factor. Maximum synergistic interaction was observed at H/L=50/50. The mixed gels exhibited excellent texture properties compared to other common food gels.     

Macromolecular Sieving by the Dormant Spore of Bacillus cereus

The threshold surface porosity in the dormant spore of Bacillus cereus strain T was assessed by measuring passive permeabilities to a series of polydisperse polyethylene glycol samples which increased in average molecular size. The apparent exclusion threshold at diffusional equilibrium corresponded to a polymer of number-average molecular weight ( M(n)) = 150,000 and equivalent hydrodynamic radius ( r(ES)) = 16 nm, which confirmed a previous report. However, analytical gel chromatography before and after uptake by the spores revealed that only the low molecular weight fractions in a polymer sample distribution were taken up. From graphical analyses of the changes in molecular weight distributions, a quasi-monodisperse exclusion threshold was determined corresponding to M(n) = 8,000 and r(ES) = 3.2 nm. Thus, the equivalent porosity in the limiting outer integument appeared much more restrictive than heretofore shown for spores, although still more open than the monodisperse equivalent for the cell wall of vegetative bacilli.     

Functionalized polycarbonate derived from tartaric acid: enzymatic ring-opening polymerization of a seven-membered cyclic carbonate

Enantiomerically pure functional polycarbonate was synthesized from a novel seven-membered cyclic carbonate monomer derived from naturally occurring L-tartaric acid. The monomer was synthesized in three steps and screened for polymerization with four commercially available lipases from different sources at 80 degrees C, in bulk. The ring-opening polymerization (ROP) was affected by the source of the enzyme; the highest number-average molecular weight, M(n) = 15500 g/mol (PDI = 1.7; [alpha]D(20) = +77.8, T(m) = 58.8 degrees C) optically active polycarbonate was obtained with lipase Novozyme-435. The relationship between monomer conversion, reaction time, molecular weight, and molecular weight distribution were investigated for Novozyme-435 catalyzed ROP. Deprotection of the ketal groups was achieved with minimal polymer chain cleavage (M(n) = 10000 g/mol, PDI = 2.0) and resulted in optically pure polycarbonate ([alpha]D(20) = +56) bearing hydroxy functional groups. Deprotected poly(ITC) shows T(m) of 60.2 degrees C and DeltaH(f) = 69.56 J/g and similar to that of the poly(ITC), a glass transition temperature was not found. The availability of the pendant hydroxyl group is expected to enhance the biodegradability of the polymer and serves in a variety of potential biomedical applications such as polymeric drug delivery systems.     

Polyethyleneimine derivative for nucleic acid model

Water-soluble polyethyleneimine (PE) derivatives containing nucleic acid bases and hydrophilic amino acids such as homoserine (Hse) and serine were prepared by the activated ester method as nucleic acid models. From spectroscopic measurements, the polymers were found to interact with DNA accompanied by an induction of conformational change. Hypochromicity in UV spectra indicated that a stable polymer complex was formed between poly (A) with PEI-Hse-Ura by complementary hydrogen bonding with equimolar nucleic base units (adenine∶uracil=1∶1). The induced conformation of DNA by the interaction with the polymer containing uracil and homoserine (PEI-Hse-Ura) was concluded to be a super triple helical structure. The formation of the polymer complex, DNA:PEI-Hse-Ura, was found to be affected by the presence of metal ions such as Ca²⁺ and Cu²⁺.     

Poly(ethylene glycol)-modification of the phospholipid vesicles by using the spontaneous incorporation of poly(ethylene glycol)-lipid into the vesicles

The critical micelle concentrations of 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[monomethoxy poly(ethylene glycol) (5000)] (PEG-DPPE) and its distearoyl analogue (PEG-DSPE) were 70 and 9 microM, respectively, in buffer solutions ([Tris] = 20 mM, [NaCl] = 140 mM, pH 7.4) at 37 degrees C. When these PEG-lipid micelle dispersions were mixed with the dispersions of phospholipid vesicles comprised of a C16 membrane, of which the carbon number is 16, or a C18 membrane, the PEG-lipid micelles were dissociated into monomers and then spontaneously incorporated into the surface of the preformed vesicles. The incorporation rates and the enthalpy changes during incorporation were measured with an isothermal titration microcalorimeter. The incorporation rate of PEG-DPPE was faster than that of PEG-DSPE, because the dissociation rate of the PEG-DPPE micelles was faster than that of PEG-DSPE micelles. The incorporation equilibrium constant of PEG-DSPE was larger than that of PEG-DPPE due to its slow dissociation rate from the membrane, caused by the stronger hydrophobic interaction. The combination of PEG-DSPE and the C18 membrane was the most thermodynamically stabilized pair. Furthermore, the dispersion stability of the surface-modified vesicles prepared by this spontaneous incorporation was analyzed by using the critical molecular weight of the polymer for the aggregation of vesicles. The aggregation of the vesicles was successfully supressed with an increase in the molecular weight of the PEG in the PEG-lipid and its incorporation ratio.     

Molecular dynamics study of the influence of solvents on the structure and mechanical properties of poly(vinyl alcohol) gels

Molecular dynamics (MD) simulations were employed to study the influence of solvents on the structure and mechanical properties of physically crosslinked poly(vinyl alcohol) (PVA) gels. Firstly, three kinds of PVA precursor gels were made by adding water, dimethyl sulfoxide (DMSO) and a mixture of DMSO and water (4:1 by weight), respectively. The solvents in the precursor gels were then exchanged with water to obtain three kinds of PVA hydrogels. Solvent in the precursor gel with a mixture of DMSO and water was also exchanged with ethanol and DMSO, respectively. It was found that the tensile strength and failure strain of the PVA hydrogel prepared from precursor gel with a mixture of DMSO and water was the highest, and the polymer network was more homogeneous than the other two PVA hydrogels. The polymer network of PVA gel with ethanol or with DMSO was more heterogenous than with water, and the tensile strength and failure strain were much lower. The torsional activity of polymer chains of PVA gel with ethanol was much stronger than PVA gel with water and DMSO.     

Preparation and copolymerization of a novel carbohydrate containing monomer

A novel carbohydrate containing monomer was prepared by simple reaction of 2-vinyl-4,4′-dimethylazlactone (VDMA) and 1,2;5,6-diisopropylidene--d-glucofuranose (DAG). The monomer was easily homopolymerized as well as copolymerized with methyl methacrylate (MMA) to give high molecular weight polymers using free radical conditions. Upon removal of the isopropylidene groups from the polymer, water contact angles decreased for polymeric films of the material, indicating a more hydrophilic surface. The addition of a carbohydrate moiety to MMA copolymers increases its hydrophilicity and allows for potential use of the new polymer as a biomaterial in a variety of applications.