1,185 research outputs found
Synthesis of Fluorine-18 Functionalized Nanoparticles for use as in vivo Molecular Imaging Agents
Nanoparticles containing fluorine-18 were prepared from block copolymers made by ring opening metathesis polymerization (ROMP). Using the fast initiating ruthenium metathesis catalyst (H_2IMes)(pyr)_2(Cl)_2Ru=CHPh, low polydispersity amphiphilic block copolymers were prepared from a cinnamoyl-containing hydrophobic norbornene monomer and a mesyl-terminated PEG-containing hydrophilic norbornene monomer. Self-assembly into micelles and subsequent cross-linking of the micelle cores by light-activated dimerization of the cinnamoyl groups yielded stable nanoparticles. Incorporation of fluorine-18 was achieved by nucleophilic displacement of the mesylates by the radioactive fluoride ion with 31% incorporation of radioactivity. The resulting positron-emitting nanoparticles are to be used as in vivo molecular imaging agents for use in tumor imaging
Degradation versus self-assembly of block copolymer micelles
The stability of micelles self-assembled from block copolymers can be altered
by the degradation of the blocks. Slow degradation shifts the equilibrium size
distribution of block copolymer micelles and change their properties.
Quasi-equilibrium scaling theory shows that the degradation of hydrophobic
blocks in the core of micelles destabilize the micelles reducing their size,
while the degradation of hydrophilic blocks forming coronas of micelles favors
larger micelles and may, at certain conditions, induce the formation of
micelles from individual chains.Comment: Published in Langmuir http://pubs.acs.org/doi/pdf/10.1021/la204625
Quantum dot loaded immunomicelles for tumor imaging
<p>Abstract</p> <p>Background</p> <p>Optical imaging is a promising method for the detection of tumors in animals, with speed and minimal invasiveness. We have previously developed a lipid coated quantum dot system that doubles the fluorescence of PEG-grafted quantum dots at half the dose. Here, we describe a tumor-targeted near infrared imaging agent composed of cancer-specific monoclonal anti-nucleosome antibody 2C5, coupled to quantum dot (QD)-containing polymeric micelles, prepared from a polyethylene glycol/phosphatidylethanolamine (PEG-PE) conjugate. Its production is simple and involves no special equipment. Its imaging potential is great since the fluorescence intensity in the tumor is twofold that of non-targeted QD-loaded PEG-PE micelles at one hour after injection.</p> <p>Methods</p> <p>Para-nitrophenol-containing (5%) PEG-PE quantum dot micelles were produced by the thin layer method. Following hydration, 2C5 antibody was attached to the PEG-PE micelles and the QD-micelles were purified using dialysis. 4T1 breast tumors were inoculated subcutaneously in the flank of the animals. A lung pseudometastatic B16F10 melanoma model was developed using tail vein injection. The contrast agents were injected via the tail vein and mice were depilated, anesthetized and imaged on a Kodak Image Station. Images were taken at one, two, and four hours and analyzed using a methodology that produces normalized signal-to-noise data. This allowed for the comparison between different subjects and time points. For the pseudometastatic model, lungs were removed and imaged <it>ex vivo </it>at one and twenty four hours.</p> <p>Results</p> <p>The contrast agent signal intensity at the tumor was double that of the passively targeted QD-micelles with equally fast and sharply contrasted images. With the side views of the animals only tumor is visible, while in the dorsal view internal organs including liver and kidney are visible. <it>Ex vivo </it>results demonstrated that the agent detects melanoma nodes in a lung pseudometastatic model after a 24 hours wash-out period, while at one hour, only a uniform signal is detected.</p> <p>Conclusions</p> <p>The targeted agent produces ultrabright tumor images and double the fluorescence intensity, as rapidly and at the same low dose as the passively targeted agents. It represents a development that may potentially serve to enhance early detection for metastases.</p
A 13C-NMR study of the role of Asn-155 in stabilizing the oxyanion of a subtilisin tetrahedral adduct
Enhancing Specific Disruption of Intracellular Protein Complexes by Hydrocarbon Stapled Peptides Using Lipid Based Delivery
Linear peptides can mimic and disrupt protein-protein interactions involved in critical cell signaling pathways. Such peptides however are usually protease sensitive and unable to engage with intracellular targets due to lack of membrane permeability. Peptide stapling has been proposed to circumvent these limitations but recent data has suggested that this method does not universally solve the problem of cell entry and can lead to molecules with off target cell lytic properties. To address these issues a library of stapled peptides was synthesized and screened to identify compounds that bound Mdm2 and activated cellular p53. A lead peptide was identified that activated intracellular p53 with negligible nonspecific cytotoxicity, however it still bound serum avidly and only showed a marginal improvement in cellular potency. These hurdles were overcome by successfully identifying a pyridinium-based cationic lipid formulation, which significantly improved the activity of the stapled peptide in a p53 reporter cell line, principally through increased vesicular escape. These studies under score that stapled peptides, which are cell permeable and target specific, can be identified with rigorous experimental design and that these properties can be improved through use with lipid based formulations. This work should facilitate the clinical translation of stapled peptides
Small Angle Scattering and Zeta Potential of Liposomes Loaded with Octa(carboranyl)porphyrazine
In this work the physicochemical characterization of liposomes loaded with a
newly synthesised carboranyl porphyrazine (H2HECASPz) is described. This
molecule represents a potential drug for different anticancer therapies, such
as Boron Neutron Capture Therapy, Photodynamic Therapy and Photothermal
Therapy. Different loading methods and different lipid mixtures were tested.
The corresponding loaded vectors were studied by Small Angle Scattering (SANS
and SAXS), light scattering and zeta potential. The combined analysis of
structural data at various length scales and the measurement of the surface
charge allowed to obtain a detailed characterization of the investigated
systems. The mechanisms underlying the onset of differences in relevant
physicochemical parameters (size, polydispersity and charge) were also
critically discussed
Effect of Liposome Characteristics and Dose on the Pharmacokinetics of Liposomes Coated with Poly(amino acid)s
Long-circulating liposomes, such as PEG-liposomes, are frequently studied for drug delivery and diagnostic purposes. In our group, poly(amino acid) (PAA)-based coatings for long-circulating liposomes have been developed. These coatings provide liposomes with similar circulation times as compared to PEG-liposomes, but have the advantage of being enzymatically degradable. For PEG-liposomes it has been reported that circulation times are relatively independent of their physicochemical characteristics. In this study, the influence of factors such as PAA grafting density, cholesterol inclusion, surface charge, particle size, and lipid dose on the circulation kinetics of PAA-liposomes was evaluated after intravenous administration in rats. Prolonged circulation kinetics of PAA-liposomes can be maintained upon variation of liposome characteristics and the lipid dose given. However, the use of relatively high amounts of strongly charge-inducing lipids and a too large mean size is to be avoided. In conclusion, PAA-liposomes represent a versatile drug carrier system for a wide variety of applications
Catechol Polymers for pH-Responsive, Targeted Drug Delivery to Cancer Cells
A novel cell-targeting, pH-sensitive polymeric carrier was employed in this study for delivery of the anticancer drug bortezomib (BTZ) to cancer cells. Our strategy is based on facile conjugation of BTZ to catechol-containing polymeric carriers that are designed to be taken up selectively by cancer cells through cell surface receptor-mediated mechanisms. The polymer used as a building block in this study was poly(ethylene glycol), which was chosen for its ability to reduce nonspecific interactions with proteins and cells. The catechol moiety was exploited for its ability to bind and release borate-containing therapeutics such as BTZ in a pH-dependent manner. In acidic environments, such as in cancer tissue or the subcellular endosome, BTZ dissociates from the polymer-bound catechol groups to liberate the free drug, which inhibits proteasome function. A cancer-cell-targeting ligand, biotin, was presented on the polymer carriers to facilitate targeted entry of drug-loaded polymer carriers into cancer cells. Our study demonstrated that the cancer-targeting drug-polymer conjugates dramatically enhanced cellular uptake, proteasome inhibition, and cytotoxicity toward breast carcinoma cells in comparison with nontargeting drug-polymer conjugates. The pH-sensitive catechol-boronate binding mechanism provides a chemoselective approach for controlling the release of BTZ in targeted cancer cells, establishing a concept that may be applied in the future toward other boronic acid-containing therapeutics to treat a broad range of diseases
Self-assembled nanogel made of mannan : synthesis and characterization
Amphiphilic mannan (mannan-C16) was synthesized by the Michael addition of hydrophobic 1-hexadecanethiol (C16)
to hydroxyethyl methacrylated mannan (mannan-HEMA). Mannan-C16 formed nanosized aggregates in water by selfassembly
via the hydrophobic interaction among C16molecules as confirmed by hydrogen nuclearmagnetic resonance (1H
NMR), fluorescence spectroscopy, cryo-field emission scanning electron microscopy (cryo-FESEM), and dynamic light
scattering (DLS). The mannan-C16 critical aggregation concentration (cac), calculated by fluorescence spectroscopy with
Nile red and pyrene, ranged between 0.04 and 0.02mg/mL depending on the polymer degree of substitution ofC16 relative
to methacrylated groups. Cryo-FESEM micrographs revealed that mannan-C16 formed irregular spherical macromolecular
micelles, in this work designated as nanogels, with diameters ranging between 100 and 500 nm. The influence of the
polymer degree of substitution, DSHEMA andDSC16, on the nanogel size and zeta potential was studied byDLS at different
pH values and ionic strength and as a function of mannan-C16 and urea concentrations. Under all tested conditions, the
nanogel was negatively charged with a zeta potential close to zero. Mannan-C16 with higher DSHEMA and DSC16
values
formed larger nanogels andwere also less stable over a 6month storage period and at concentrations close to the cac.When
exposed to solutions of different pH and aggressive conditions of ionic strength and urea concentration, the size of
mannan-C16 varied to some extent but was always in the nanoscale range.International Iberian
Nanotechnology Laboratory (INL)Fundação para a Ciência e a Tecnologia (FCT
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