温度敏感的PLGA-PEG-PLGA水凝胶的合成、表征和药物释放

用聚乙二醇PEG1000和4600引发乙交酯(GA)和L-丙交酯(L-LA)开环共聚合得到一系列数均分子量为3 000~7 000的PLGA-PEG-PLGA水凝胶材料.综合应用动态粘弹谱仪和相图,系统报道了该凝胶力学性质和溶胶-凝胶转变的关系,凝胶区间的模量在102~104Pa之间.用荧光光谱证明了该三嵌段聚合物形成胶束的性质并测定了临界胶束浓度,验证了凝胶由胶束形成的机理.凝胶中的头孢他定释放呈现一定程度的缓释作用.

Gene delivery using dimethyldidodecylammonium bromide-coated PLGA nanoparticles

In this present work we describe a poly(lactic-co-glycolic acid) (PLGA) nanoparticle formulation for intracellular delivery of plasmid DNA. This formulation was developed to encapsulate DNA within PLGA nanoparticles that combined salting out and emulsion evaporation processes. This process reduced the requirement for sonication which can induce degradation of the DNA. A monodispersed nanoparticle population with a mean diameter of approximately 240 nm was produced, entrapping a model plasmid DNA in both supercoiled and open circular structures. To induce endosomal escape of the nanoparticles, a superficial cationic charge was introduced using positively charged surfactants cetyl trimethylammonium bromide (CTAB) and dimethyldidodecylammonium bromide (DMAB), which resulted in elevated zeta potentials. As expected, both cationic coatings reduced cell viability, but at equivalent positive zeta potentials, the DMAB coated nanoparticles induced significantly less cytotoxicity than those coated with CTAB. Fluorescence and transmission electron microscopy demonstrated that the DMAB coated cationic nanoparticles were able to evade the endosomal lumen and localise in the cytosol of treated cells. Consequently, DMAB coated PLGA nanoparticles loaded with a GFP reporter plasmid exhibited significant improvements in transfection efficiencies with comparison to non-modified particles, highlighting their functional usefulness. These nanoparticles may be useful in delivery of gene therapies to targeted cells. (C) 2010 Elsevier Ltd. All rights reserved.

Antibody Targeting of Camptothecin-Loaded PLGA Nanoparticles to Tumor Cells

Antibody targeting of drug substances can improve the efficacy of the active molecule, improving distribution and concentration of the drug at the site of injury/disease. Encapsulation of drug substances into polymeric nanoparticles can also improve the therapeutic effects of such compounds by protecting the molecule until its action is required. In this current study, we have brought together these two rationales to develop a novel immunonanoparticle with improved therapeutic effect against colorectal tumor cells. This nanoparticle comprised a layer of peripheral antibodies (Ab) directed toward the Fas receptor (CD95/Apo-1) covalently attached to poly(lactide-co-glycolide) nanoparticles (NP) loaded with camptothecin. Variations in surface carboxyl density permitted up to 48.5 mu g coupled Ab per mg of NP and analysis of nanoparticulate cores showed efficient camptothecin loading. Fluorescence visualization studies confirmed internalization of nanoconstructs into endocytic compartments of HCT 116 cells, an effect not evident in NP without superficial Ab. Cytotoxicity studies were then carried out against HCT116 cells. After 72 h, camptothecin solution resulted in an IC50 of 21.8 ng mL(-1). Ab-directed delivery of NP-encapsulated camptothecin was shown to be considerably more effective with an IC50 of 0.37 ng mL(-1). Calculation of synergistic ratios for these nanoconstructs demonstrated synergy of pharmacological relevance. Indeed, the results in this paper suggest that the attachment of anti-Fas antibodies to camptothecin-loaded nanoparticles may result in a therapeutic strategy that could have potential in the treatment of tumors expressing death receptors.

New grafted PLA-g-PEG polymeric nanoparticles used to improve bioavailability of oral drugs.

Les nanoparticules (NPs) de polymère ont montré des résultats prometteurs pour leur utilisation comme système de transport de médicaments pour une libération contrôlée du médicament, ainsi que pour du ciblage. La biodisponibilité des médicaments administrés oralement pourrait être limitée par un processus de sécrétion intestinale, qui pourrait par la suite être concilié par la glycoprotéine P (P-gp). La dispersion de la Famotidine (modèle de médicament) à l’intérieur des nanoparticules (NPs) pegylées a été évaluée afin d’augmenter la biodisponibilité avec du polyéthylène glycol (PEG), qui est connu comme un inhibiteur de P-gp. L’hypothèse de cette étude est que l’encapsulation de la Famotidine (un substrat de P-gp) à l’intérieur des NPs préparées à partir de PEG-g-PLA pourrait inhiber la fonction P-gp. La première partie de cette étude avait pour but de synthétiser quatre copolymères de PEG greffés sur un acide polylactide (PLA) et sur un squelette de polymère (PLA-g-PEG), avec des ratios de 1% et 5% (ratio molaire de PEG vs acide lactique monomère) de soit 750, soit 2000 Da de masse moléculaire. Ces polymères ont été employés afin de préparer des NPs chargés de Famotidine qui possède une faible perméabilité et une solubilité aqueuse relativement basse. Les NPs préparées ont été analysées pour leur principaux paramètres physicochimiques tels que la taille et la distribution de la taille, la charge de surface (Potentiel Zeta), la morphologie, l’efficacité d’encapsulation, le pourcentage résiduel en alcool polyvinylique (PVA) adsorbé à la surface des NPs, les propriétés thermiques, la structure cristalline et la libération du médicament. De même, les formules de NPs ont été testées in vitro sur des cellules CaCo-2 afin dʼévaluer la perméabilité bidirectionnelle de la Famotidine. Généralement, les NPs préparées à partir de polymères greffés PLA-g-5%PEG ont montré une augmentation de la perméabilité du médicament, ce par l’inhibition de l’efflux de P-gp de la Famotidine dans le modèle CaCo-2 in vitro. Les résultats ont montré une baisse significative de la sécrétion de la Famotidine de la membrane basolatéral à apical lorsque la Famotidine était encapsulée dans des NPs préparées à partir de greffes de 5% PEG de 750 ou 2000 Da, de même que pour d’autres combinaisons de mélanges physiques contenant du PEG5%. La deuxième partie de cette étude est à propos de ces NPs chargées qui démontrent des résultats prometteurs en termes de perméabilité et d’inhibition d’efflux de P-gp, et qui ont été choises pour développer une forme orale solide. La granulation sèche a été employée pour densifier les NPs, afin de développer des comprimés des deux formules sélectionnées de NPs. Les comprimés à base de NPs ont démontré un temps de désintégration rapide (moins d’une minute) et une libération similaire à la Famotidine trouvée sur le marché. Les résultats de l’étude du transport de comprimés à base de NPs étaient cohérents avec les résultats des formules de NPs en termes d’inhibition de P-gp, ce qui explique pourquoi le processus de fabrication du comprimé n’a pas eu d’effet sur les NPs. Mis ensemble, ces résultats montrent que l’encapsulation dans une NP de polymère pegylé pourrait être une stratégie prometteuse pour l’amélioration de la biodisponibilité des substrats de P-gp.

Effect of Polymer Architecture on the Structural and Biophysical Properties of PEG-PLA Nanoparticles

Polymers made of poly(ethylene glycol) chains grafted to poly(lactic acid) chains (PEG-g-PLA) were used to produce stealth drug nanocarriers. A library of comb-like PEG-g-PLA polymers with different PEG grafting densities was prepared in order to obtain nanocarriers with dense PEG brushes at their surface, stability in suspension, and resistance to protein adsorption. The structural properties of nanoparticles (NPs) produced from these polymers by a surfactant-free method were assessed by DLS, zeta potential, and TEM and were found to be controlled by the amount of PEG present in the polymers. A critical transition from a solid NP structure to a soft particle with either a “micelle-like” or “polymer nano-aggregate” structure was observed when the PEG content was between 15 to 25% w/w. This structural transition was found to have a profound impact on the size of the NPs, their surface charge, their stability in suspension in presence of salts as well as on the binding of proteins to the surface of the NPs. The arrangement of the PEG-g-PLA chains at the surface of the NPs was investigated by 1H NMR and X-ray photoelectron spectroscopy (XPS). NMR results confirmed that the PEG chains were mostly segregated at the NP surface. Moreover, XPS and quantitative NMR allowed quantifying the PEG chain coverage density at the surface of the solid NPs. Concordance of the results between the two methods was found to be remarkable. Physical-chemical properties of the NPs such as resistance to aggregation in saline environment as well as anti-fouling efficacy were related to the PEG surface density and ultimately to polymer architecture. Resistance to protein adsorption was assessed by isothermal titration calorimetry (ITC) using lysozyme. The results indicate a correlation between PEG surface coverage and level of protein interactions. The results obtained lead us to propose such PEG-g-PLA polymers for nanomedecine development as an alternative to the predominant polyester-PEG diblock polymers.

Zidovudine-Loaded PLA and PLA-PEG Blend Nanoparticles: Influence of Polymer Type on Phagocytic Uptake by Polymorphonuclear Cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of praziquantel-loaded PLGA nanoparticles and evaluation of intestinal permeation by the everted gut Sac model

Praziquantel has been shown to be highly effective against all known species of Schistosoma infecting humans. Spherical nanoparticles made of poly(D,L-lactide-co-glycolide) acid with controlled size were designed as drug carriers. Praziquantel, a hydrophobic drug, was entrapped into the polymeric nanoparticles with 30% (w/w) of theoretical loading. The nanoparticles size was approximately of 350 nm with 66% of encapsulation efficiency. The everted gut sac model shows to be efficient to evaluate the drug permeation through the intestinal membrane. The results show that free praziquantel presents 4-fold times more permeation than praziquantel-loaded PLGA nanoparticles and physical mixture. For this drug, in special, this result can be interesting, since the nanoparticulate system can behave as a drug reservoir and/or to have a more localized effect in intestinal membrane for a prolonged period of time, since great amounts of parasites can be usually found in the mesenteric veins.

Praziquantel-loaded PLGA nanoparticles: preparation and characterization

Nanoparticles containing praziquantel made of Poly (D,L-lactide-co-glycolide) were designed by an emulsion-solvent evaporation method. Two organic solvents were separately utilized as disperse phase: methylene chloride and ethyl acetate. The size of the particles prepared with the former solvent was bigger than the particles prepared with the latter. The entrapment efficiency was bigger when methylene chloride was used, 79.82% in comparison with 29.27% by using ethyl acetate. DSC and infrared studies showed that no strong chemical interaction between drug and polymer occurred. Release kinetics of praziquantel, used as a model drug, was governed not only by actual drug loading but also by particles size. The higher the drug content and the smaller the particle size resulted in faster drug release.

PLGA nanoparticles containing praziquantel: effect of formulation variables on size distribution

Praziquantel has been shown to be highly effective against all known species of Schistosoma infecting humans. Spherical nanoparticulate drug carriers made of poly(D,L-lactide-co-glycolide) acid with controlled size were designed. Praziquantel, a hydrophobic molecule, was entrapped into the nanoparticles with theoretical loading varying from 10 to 30% (w/w). This investigates the effects of some process variables on the size distribution of nanoparticles prepared by emulsion-solvent evaporation method. The results show that sonication time, PLGA and drug amounts, PVA concentration, ratio between aqueous and organic phases, and the method of solvent evaporation have a significant influence on size distribution of the nanoparticles. (C) 2004 Elsevier B.V. All rights reserved.

Intranasal delivery of zidovudine by PLA and PLA-PEG blend nanoparticles

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Thermoanalytical study of praziquantel-loaded PLGA nanoparticles

Polymeric nanoparticles have received great attention as potential controlled drug delivery systems. Biodegradable polymers has been extensively used in the development of these drug carriers, and the polyesters such as polylactic acid, polyglycolic acid and their copolymers as poly-lactide-co- glycolide are the most used, considering its biocompatibility and biodegradability. Thermal analysis techniques have been used for pharmaceutical substances for more than 30 years and are routine methods for screening drug-excipient interactions. The aim of this work is to use thermal analysis to characterize PLGA nanoparticles containing a hydrophobic drug, praziquantel. The results show that the drug is in an amorphous state or in disordered crystalline phase of molecular dispersion in the PLGA polymeric matrix and that the microencapsulation process did not interfere with the chemical structure of the polymer, mantaining the structural drug integrity.

Antioxidant Effects of Quercetin and Catechin Encapsulated into PLGA Nanoparticles

Polymeric nanoparticles (PLGA) have been developed for the encapsulation and controlled release of quercetin and catechin. Nanoparticles were fabricated using a solvent displacementmethod. Physicochemical properties were measured by light scattering, scanning electron microscopy and zeta-potential, X-ray diffraction, infrared spectroscopy and differential scanning calorimetry. Encapsulation efficiency and in vitro release profiles were obtained from differential pulse voltammetry experiments. Antioxidant properties of free and encapsulated flavonoids were determined by TBARS, fluorescence spectroscopy and standard chelating activity methods. Relatively small (d approximate to 400 nm) polymeric nanoparticles were obtained containing quercetin or catechin in a non-crystalline form (EE approximate to 79%) and the main interactions between the polymer and each flavonoid were found to consist of hydrogen bonds. In vitro release profiles were pH-dependant, the more acidic pH, the faster release of each flavonoid from the polymeric nanoparticles. The inhibition of the action of free radicals and chelating properties, were also enhanced when quercetin and catechin were encapsulated within PLGA nanoparticles. The information obtained from this study will facilitate the design and fabrication of polymeric nanoparticles as possible oral delivery systems for encapsulation, protection and controlled release of flavonoids aimed to prevent oxidative stress in human body or food products.

Curcumin-loaded into PLGA nanoparticles

The incorporation of the curcumin into poly(lactic-co-glycolic)acid (PLGA) nanospheres by the nanoprecipitation technique, the characterization of the nanoparticles and the schistosomicidal activity of the curcumin-loaded into PLGA nanospheres were reported. The incorporation process occurred with high efficiency and the images of field-emission scanning electron microscopy (FESEM) revealed the production of spherically shaped particles. According to the dynamic light scattering measurements, the particles are nanometric and monodisperse. The curcumin-loaded PLGA nanoparticles (50 and 100 mu M) caused the death of all worms and a separation between 50% and 100% of Schistosoma mansoni couples at concentrations from 30 mu M. Moreover, the curcumin-loaded PLGA nanoparticles also decreased the motor activity and caused partial alterations in the tegument of adult worms. This study marks the first time that schistosomicidal activity has been reported for curcumin-loaded PLGA nanoparticles.

Cationic Bovine Serum Albumin (CBA) conjugated Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles for extended delivery of methotrexate into brain tumor

Current treatment strategies for the treatment of brain tumor have been hindered primarily by the presence of highly lipophilic insurmountable blood-brain barrier (BBB). The purpose of current research was to investigate the efficiency of engineered biocompatible polymeric nanoparticles (NPs) as drug delivery vehicle to bypass the BBB and enhance biopharmaceutical attributes of anti-metabolite methotrexate (MTX) encapsulated NPs. The NPs were prepared by solvent diffusion method using cationic bovine serum albumin (CBA), and characterized for physicochemical parameters such as particle size, polydispersity index, and zeta-potential; while the surface modification was confirmed by FTIR, and NMR spectroscopy. Developed NPs exhibited zestful relocation of FITC tagged NPs across BBB in albino rats. Further, hemolytic studies confirmed them to be non-toxic and biocompatible as compared to free MTX. In vitro cytotoxicity assay of our engineered NPs on HNGC1 tumor cells proved superior uptake in tumor cells; and elicited potent cytotoxic effect as compared to plain NPs and free MTX solution. The outcomes of the study evidently indicate the prospective of CBA conjugated poly (D,L-lactide-co-glycolide) (PLGA) NPs loaded with MTX in brain cancer bomber with amplified capability to circumvent BBB.

Preparation and in vivo evaluation of insulin-loaded biodegradable nanoparticles prepared from diblock copolymers of PLGA and PEG

The aim of this study was to design a controlled release vehicle for insulin to preserve its stability and biological activity during fabrication and release. A modified, double emulsion, solvent evaporation, technique using homogenisation force optimised entrapment efficiency of insulin into biodegradable nanoparticles (NP) prepared from poly (dl-lactic-co-glycolic acid) (PLGA) and its PEGylated diblock copolymers. Formulation parameters (type of polymer and its concentration, stabiliser concentration and volume of internal aqueous phase) and physicochemical characteristics (size, zeta potential, encapsulation efficiency, in vitro release profiles and in vitro stability) were investigated. In vivo insulin sensitivity was tested by dietinduced type II diabetic mice. Bioactivity of insulin was studied using Swiss TO mice with streptozotocin-induced type I diabetic profile. Insulin-loaded NP were spherical and negatively charged with an average diameter of 200–400 nm. Insulin encapsulation efficiency increased significantly with increasing ratio of co-polymeric PEG. The internal aqueous phase volume had a significant impact on encapsulation efficiency, initial burst release and NP size. Optimised insulin NP formulated from 10% PEG-PLGA retained insulin integrity in vitro, insulin sensitivity in vivo and induced a sustained hypoglycaemic effect from 3 hours to 6 days in type I diabetic mice.