Pharmaceutical Nanotechnology (v.1, #3)

Editorial: by Ijeoma Uchegbu (163-163).

Spotlight: Stefaan De Smedt by Kevin Braeckmans (164-164).

Current Trends in the Use of Cationic Polymer Assemblies for siRNA and Plasmid DNA Delivery by Gemma Navarro, Sara Movassaghian, Vladimir P. Torchilin (165-183).
Gene therapy is a promising approach for disease prevention and therapy. Efficient gene delivery is an importantfactor limiting gene delivery. Due to the poor stability of nucleotide molecules in vivo, they require an associationwith delivery systems to overcome extracellular and intracellular barriers and allow access to the site of action. In this review,we discuss the challenges currently encountered in the delivery of DNA and RNAi-based therapies using the mostcommon cationic polymer assemblies including polylysines, chitosan, polyethylenimines, dendrimers, cyclodextrins andillustrate with examples how chemical modification of cationic assemblies may contribute to their successful applicationin the clinic.

Folic Acid Conjugated Chitosan Nanoparticles for Tumor Targeting of Therapeutic and Imaging Agents by Driton Vllasaliu, Luca Casettari, Giulia Bonacucina, Marco Cespi, Giovanni Filippo Palmieri, Lisbeth Illum (184-203).
Anticancer drugs are typically distributed non-specifically in the body, where they affect both cancerous andnormal cells. This limits the drug level achievable within the tumor, compromising the therapeutic efficacy, and results inpotential toxic effects on normal tissues. Targeted delivery of chemotherapeutics exclusively to cancer cells is the focus ofintensive research for improvement of anticancer therapy. Various drug delivery systems have been investigated for thispurpose, with therapeutic-carrying polymeric nanoparticulate systems designed for specific targeting of tumor cells receivingspecial interest. Chitosan, a natural polymer derived from crustacean shells, has attracted particular attention as adrug carrier. The simple and mild preparation methods, low toxicity, good stability, controlled drug release and the abilityto overcome biological barriers have made chitosan-based nanoparticles popular in drug and gene delivery applications.Chitosan nanoparticles have been fabricated with optimal size and surface characteristics in order to tailor the behaviorwithin the biological system, including circulation time, as well as passive and active cancer targeting. Folic acid is widelyemployed as a ligand targeting cancerous cells as its receptor which ‘shuttles’ folic acid into the cells via endocytosis isover-expressed on the surface of many human epithelial cancer cells. Incorporating folic acid into chitosan-based drug andgene delivery formulations renders the systems with an efficient targeting capacity. Furthermore, it is possible to formulatechitosan nanocarriers that display multiple useful characteristics extending beyond targeted delivery. The versatilityof these systems is also being exploited in nanotheranostics.

The present work explores the preparation and characterization of chitosan/lecithin nanoparticles loaded witharteether (ART) and arteether entrapped in cyclodextrins (ART-β CD) to boost its anti-malarial activity. Arteether, an antimalarialdrug was chosen by the Steering Committee of the Scientific Working Group on Malaria Chemotherapy of theWHO (CHEMAL) for treatment of cerebral malaria. Unfortunately, ART is water-insoluble drug (17 μg/ml at roomtemp.) and its therapeutic efficacy is greatly hampered due to poor bioavailability (~40%, degradation in stomach acids).Moreover, Arteether have short plasma half-life which requires frequent administration. Formation of nanoparticles ofART can be a suitable solution to improve their Biopharmaceutical properties. The nanoparticles prepared using modifiedsolvent evaporation method depicted a particle size in the range of 299-354 nm for arteether and 157-212 nm for ART-βCD loaded nanoparticles. 100 mg loaded ART and ART-β CD formulations showed maximum drug entrapment efficiency.Prepared nanoparticles reflected spherical shape inTEM images. Disappearance of decomposition endotherm inDSC scans of nanoparticles revealed the increased physical stability. FT-IR spectra showed small changes in major peaksof drug negating any chemical change in the drug when entrapped in the nanoparticle formulation. In vitro drug releasestudies suggested the controlled release as well as improved pattern. Enhanced antimalarial activity was observed in ARTand ART-β CD containing nanoparticles.

The use of nanovesicles for enhanced topical/transdermal delivery of therapeutic agents has been extensivelyexplored in recent years. In our previous study, nanovesicles ethosomes based gel formulation for topical delivery of anticancerdrug 5-FU was developed for the treatment of actinic keratosis and non melanoma skin cancers. The exact mechanismof better skin permeation and deposition of drugs from ethosomes is not yet fully understood. Therefore, the investigationwas aimed to understand the mechanism for better inter and intracellular drug delivery from ethosomes by quantitativeestimation of skin lipids and microscopic evaluation of nanovesicles treated skin for lipid perturbation effects. Marketed5-FU cream and drug solution were used as control for comparison purpose. Results of the biochemical estimationshowed that nanovesicles gel formulation produced maximum perturbation of skin lipids as evidenced by highest quantityof cholesterol (30.2±1.7%) and triglycerides (26.4±0.9%) extracted after 24 h from excised rat skin. In comparison, percentageof cholesterol and triglyceride extracted with marketed cream and drug solution was found to be 5.2±0.2%,2.3±0.3% and 4.4±0.1, 1.9±0.2%, respectively. Microscopic study revealed that nanovesicles gel formulation influencedthe ultra structure of the skin. Distinct regions with lamellar stacks derived from vesicles were observed in the intracellularregion of deeper skin layers. Results demonstrated that 5-FU nanovesicles gel formulation does not act only on superficiallayers of the stratum corneum, but may also induce lipid perturbations in deeper layers of the skin, whereas the marketed5-FU cream formulation remain fused on the top of stratum corneum causing an additional barrier to diffusion of thedrug. The results of the present study demonstrated that the nanovesicles can forge paths in the disordered stratum corneum,change its biochemical constituents and finally release the drug in the deeper layers of the skin.

Poly(allylamine) Magnetomicelles for Image Guided Drug Delivery by Christopher M. Barnett, Martin R. Lees, Anthony D.M. Curtis, Paul Kong Thoo Lin, Woei Ping Cheng, Clare Hoskins (224-238).
Polymeric micelles have received considerable interest for their use as drug delivery vehicles for hydrophobicdrug solubilisation. Inorganic metallic nanoparticles have already been exploited clinically in diagnostics for their contrastability, using magnetic resonance imaging. The combination of these two platforms results in a multifunctional drug carrierfor image-guided drug delivery. Here we report the synthesis and evaluation of a new class of poly(allylamine) (PAA)polymer grafted with hydrophobic oxadiazole (Ox) pendant group in a 5% molar monomer: pendant ratio. Further, thethiol-containing pendant group facilitated the attachment of hybrid iron oxide-gold nanoparticles (HNPs) via dative covalentbonding. Physicochemical characterisation of both PAA-Ox5 and PAA-Ox5-HNP polymers was carried out usingelemental analysis, nuclear magnetic resonance (NMR), fourier transform infrared spectroscopy (FTIR) and photon correlationspectroscopy (PCS). The drug loading potential of these novel aggregates was investigated, through direct conjugationof hydrophilic and encapsulation of hydrophobic drugs, respectively. The model hydrophobic drugs 2,6-diisopropylphenol (propofol) and (2S,6'R)-7-chloro-2',4,6-trimethoxy-6'-methyl-3H,4'H-spiro[1-benzofuran-2,1'-cyclohex[2]ene]-3,4'-dione (griseofulvin), and the chemotherapeutic agents bisnapthalamidopropyldiaminooctane (BNIPDaoct)and 6-Thioguanine (6-TG) were used. The data showed that the addition of HNPs onto the PAA-Ox5 structure resultedin aggregates of 175 nm in diameter. The PAA-Ox5-HNP nano-aggregates were capable of high drug solubilisationcapacities (25.79 mgmL-1, 1.68 mgmL-1 and 0.92 mgmL-1) for propofol, griseofulvin and BNIPDaoct, respectively. 6-TGwas also successfully conjugated into the polymer structure (2.8 mgmL-1). In vitro assays on human pancreatic adenocarcinomacells (BxPC-3) showed increased drug uptake and decreased IC50 values using the novel formulations comparedwith free drug. This study highlights the potential of PAA-Ox5-HNP as a bi-functional imaging and drug delivery platform.

Quatiepine, an effective atypical anti psychotic administered as fumarate or hemifumarate salts by intravenousinjection shows poor brain uptake due to low partitioning and/or Pgp efflux. In order to improve their brain availability,solid lipid nanoparticles (SLN) loaded with Quetiapine fumarate or hemifumarate were prepared using glycerylmonostearate (GMS), poloxamer 407 and hydrogenated soya phosphatidylcholine (HSPC) as stabilizers—using a hot meltemulsification high- pressure homogenization technique. They were characterized for physical characteristics like particlesize, polydispersity (PDI), shape and entrapment efficiency (EE). Fomulation and process parameters were optimizedbased on particle size, PDI and EE. SLNs with a mean particle size of 101.1 nm were obtained for quetiapine fumarateand 93.6 nm for quetiapine hemifumarate. In-vitro drug release study showed the release followed Higuchi kinetics modelfor both the formulations. In-vivo studies showed a significant increase in the percentage of drug reaching the brain whenadministered in the form of SLN’s as compared to the respective drug solutions and the increase was greater in case ofquetiapine hemifumarate salt.