Francesca Sansone

Physical characteristics and aerosol performance of naringin dry powders for pulmonary delivery prepared by spray-drying

The aim of the present work was to develop dry powders containing naringin for a direct administration to the lung to combat oxidative stress. Naringin microparticles were prepared by spray-drying the neat flavonoid (2-5% w/v) from different water/ethanol co-solvents. The spray-dried powders were characterised for morphology, density, particle size distribution, residual humidity, crystallinity, solubility, thermal behaviour and respirable fraction. The fine fraction of the powders was measured by single-stage glass impinger and Andersen cascade impactor, using the Turbospin device for the deposition tests, wherein the dose to be aerosolised was premetered in a gelatine capsule. By increasing the ethanol content, the feed liquid turned from a suspension into a solution: the spray of flavonoid suspensions led to powders with high crystallinity degree, low water solubility and high bulk density, while the spray of drug solutions led to more amorphous particles, with higher solubility, lower density and improved aerodynamic behaviour. The optimisation of the operative parameters produced enhanced aerosol performance of the flavonoid powders containing only the active compound.

Screening of a polar extract of Paeonia rockii: Composition and antioxidant and antifungal activities

ETHNOPHARMACOLOGICAL RELEVANCE The genus Paeonia (Paeoniaceae), is one of the most important source of crude drugs in traditional Chinese medicine and investigation on many species is large. Up to now studies on Paeonia rockii, one of the eight species recognized in the section Moutan, are very limited. AIM OF THE STUDY This research aimed to investigate the composition of Paeonia rockii roots and to evaluate the in vitro free-radical scavenging and antifungal activities of a polar extract (PPR) and its major constituents. MATERIALS AND METHODS PPR was obtained from defatted dried roots of Paeonia rockii using MeOH as extraction solvent. Its n-BuOH soluble portion (PPR-B) was purified by Sephadex LH-20 followed by RP-HPLC to give nineteen compounds belonging to the classes polyphenols, monoterpenes and triterpenes. Their structure were spectrally characterized (UV, 1D and 2D NMR, MS). The polyphenols content of PPR and PPR-B was examined by the Folin-Ciocalteau colorimetric assay and HPLC method. Both extracts (PPR and PPR-B) and their major constituents were tested for the free-radical scavenging activity by DPPH-test, and for the antifungal activity by three methods (micro-broth dilution method, XTT assay and Candida albicans morphological analysis). RESULTS 5-Butylhydroxy-γ-lactone (1), and ethyl-arabinopyranoside (2) have been isolated for the first time as naturally occurring compounds and taxifolin (3) was reported for the first time in Paeonia spp. Nine polyphenols, four monoterpenes and three triterpenes were also identified. Both the extracts PPR and PPR-B had high polyphenol content, and high concentration of gallic acid derivatives and paeoniflorin, chemotaxonomic characteristic markers of the genus. PPR, gallic acid and methyl-gallate displayed high potency in scavenging free-radicals (DPPH test, EC(50) 13.3, 1.2, 1.9 μg/ml, respectively). Both the extracts and gallic acid individually showed an interesting antifungal property (MIC(50) at 24 h 25, 0.9 and 30 μg/ml, respectively) and notably, a combination of paeoniflorin/gallic acid (MIC(50)=0.5+20 μg/ml, respectively) was more active than the single compound in inhibiting Candida growth. CONCLUSION The polar methanolic extract (PPR), its n-BuOH soluble fraction and constituents of Paeonia rockii were extensively investigated. Both extracts and some of their compounds have the ability to scavenge free-radicals and to inhibit Candida albicans growth.

Nanospray technology for an in situ gelling nanoparticulate powder as a wound dressing.

In the current study the feasibility of the novel nano spray drying technique for the production of stable nanoparticulate dry powder, able to gel when administered locally on a wound, is explored. Gentamicin sulphate (GS) was loaded into alginate/pectin nanoparticles as highly soluble (hygroscopic) model drug with wide range antibacterial agent for wound dressing. The influence of process variables, mainly spray mesh size and feed concentration, on particle size and morphology, powder wound fluid uptake ability and gelling rate, as well as hydrogel water vapour transmission at wound site were studied. Particles morphology was spherical with few exceptions as slightly corrugated particles when the larger nozzle was used. Production of spherical nanoparticles (d50 ∼ 350 nm) in good yield (82-92%) required 4 μm spray mesh whereas 7 μm mesh produced larger wrinkled particles. Nano spray-dried particles showed high encapsulation efficiency (∼ 80%), good flowability, high fluid uptake, fast gel formation (15 min) and proper adhesiveness to fill the wound site and to remove easily the formulation after use. Moreover, moisture transmission of the in situ formed hydrogel was between 95 and 90 g/m(2)/h, an optimum range to avoid wound dehydration or occlusion phenomena. Release of the encapsulated GS, monitored as permeation rate using Franz cells in simulated wound fluid (SWF) was related to particle size and gelling rate. Sustained permeation profiles were obtained achieving total permeation of the drug between 3 and 6 days. However, all nano spray-dried formulations presented a burst effect, suitable to prevent infection spreading at the beginning of the therapy. Antimicrobial tests against Staphylococcus aureus and Pseudomonas aeruginosa showed stronger and prolonged antimicrobial effect of the nanoparticles compared to pure GS both shortly after administration and over time (till 12 days).

Hesperidin Gastroresistant Microparticles by Spray-Drying: Preparation, Characterization, and Dissolution Profiles

Gastroresistant microparticles for oral administration of hesperidin (Hd) were produced by spray-drying using cellulose acetate phthalate (CAP) as enteric polymer in different polymer/Hd weight ratio (1:1, 3:1, and 5:1), and a series of enhancers of the dissolution rate, such as sodium carboxymethylcellulose crosslinked (CMC), sodium dodecylbenzene sulfonate (SDBS), or Tween85. The raw materials and the microparticles were investigated by differential-scanning calorimetry, X-ray diffraction, infrared spectroscopy and imaged using scanning electron and fluorescence microscopy. In vitro dissolution tests were conducted using a pH-change method to investigate the influence of formulative parameters on the dissolution/release properties of the drug. CAP/Hd microparticles showed a good gastro-resistance but incomplete drug dissolution in the simulated intestinal fluid (SIF). The presence of the enhancers in the formulation produced well-formed microparticles with different size and morphology, containing the drug well coated by the polymer. All the enhancers were able to increase the dissolution rate of Hd in the simulated intestinal environment without altering CAP ability to protect Hd in the acidic fluid. The spray-drying technique and process conditions selected were effective in microencapsulating and stabilizing the flavonoid giving satisfactory encapsulation efficiency, product yield, and microparticles morphology, and a complete drug release in the intestine.

Preparations and release characteristics of naringin and naringenin gastro-resistant microparticles by spray-drying

To prevent the drug release and degradation in gastric medium, naringin and naringenin gastro-resistant microparticles were formulated. (CAP) as coating material and spray-drying technique were tested to obtain a controlled drug release to the intestine. The influence of parameters such as the composition of the feed solution and polymer concentration on the particle yield, behavior and morphology was investigated. The microparticles were characterized by scanning electron microscopy (SEM), fluorescence microscopy (FM), and differential scanning calorimetry (DSC). In vitro dissolution studies, carried out using a pH change method, revealed that gastro-resistant naringin- and naringenin-loaded microparticles are obtainable from 2% buffer aqueous feed solutions in different polymer/drug ratios (1:1, 3:1, 5:1) by spray-drying. The method appears to be an efficient means of stabilizing these labile drugs and of obtaining a biphasic release trend due to the adequate coating properties and to the pH dependent solubility of the polymer.

Non-steroidal anti-inflammatory drug for pulmonary administration: design and investigation of ketoprofen lysinate fine dry powders.

Pulmonary inflammation is an important therapeutic target in cystic fibrosis (CF) patients, aiming to limit and delay the lung damage. The purpose of the present research was to produce respirable engineered particles of ketoprofen lysinate, a non-steroidal anti-inflammatory drug able to fight lung inflammatory status by direct administration to the site of action. Micronized drug powders containing leucine as dispersibility enhancer were prepared by co-spray drying the active compound and the excipient from water or hydro-alcoholic feeds. Microparticles were fully characterized in terms of process yield, particle size distribution, morphology and drug content. The ability of the drug to reach the deepest airways after aerosolization of spray-dried formulations was evaluated by Andersen cascade impactor, using the monodose DPI as device. In order to investigate the behaviour of the drug once in contact with lung fluid, an artificial CF mucus was prepared. Drug permeation properties were evaluated interposing the mucus layer between the drug and a synthetic membrane mounted in Franz-type diffusion cells. Finally, the effect of the engineered particles on vitality of human airway epithelial cells of patients homozygous for ΔF 508 CF (CuFi1) was studied and compared to that of raw active compound. Results indicated that powders engineering changed the diameter and shape of the particles, making them suitable for inhalation. The mucus layer in the donor compartment of vertical diffusion cells slowed down drug dissolution and permeation, leucine having no influence. Cell proliferation studies evidenced that the spray drying process together with the addition of leucine reduced the cytotoxic effect of ketoprofen lysine salt as raw material, making the ketoprofen lysinate DPI a very promising product for the inflammation control in CF patients.

Effect of flavonoids on the Aβ(25-35)-phospholipid bilayers interaction.

Amyloid-beta peptide (Abeta) is the major component of amyloid deposits found in the brain tissue of Alzheimer patients. The tendency of amyloid peptide to form amyloid plaques is known to be related to the features of the plasma membrane. Flavonoids, a group of naturally occurring molecules, exert beneficial properties to human health thanks to their antioxidant property; this property depends on their capacity to interact and permeate the cell membrane lipid bilayer. In the present research we report an Electron Paramagnetic Resonance (EPR) investigation of 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) membranes interacting with the beta-amyloid fragment Abeta(25-35), in the presence of flavonoids rutin, quercetin, naringin and naringenin. Our results, evidencing a flavonoid-dependent rigidifying effect of the bilayer, may provide the molecular basis to explain the known neuroprotective effect of flavonoid compounds.

Hazelnut (Corylus avellana L.) Shells Extract: Phenolic Composition, Antioxidant Effect and Cytotoxic Activity on Human Cancer Cell Lines

Hazelnut shells, a by-product of the kernel industry processing, are reported to contain high amount of polyphenols. However, studies on the chemical composition and potential effects on human health are lacking. A methanol hazelnut shells extract was prepared and dried. Our investigation allowed the isolation and characterization of different classes of phenolic compounds, including neolignans, and a diarylheptanoid, which contribute to a high total polyphenol content (193.8 ± 3.6 mg of gallic acid equivalents (GAE)/g of extract). Neolignans, lawsonicin and cedrusin, a cyclic diarylheptanoid, carpinontriol B, and two phenol derivatives, C-veratroylglycol, and β-hydroxypropiovanillone, were the main components of the extract (0.71%–2.93%, w/w). The biological assays suggested that the extract could be useful as a functional ingredient in food technology and pharmaceutical industry showing an in vitro scavenging activity against the radical 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) (EC50 = 31.7 μg/mL with respect to α-tocopherol EC50 = 10.1 μg/mL), and an inhibitory effect on the growth of human cancer cell lines A375, SK-Mel-28 and HeLa (IC50 = 584, 459, and 526 μg/mL, respectively). The expression of cleaved forms of caspase-3 and poly(ADP-ribose) polymerase-1 (PARP-1) suggested that the extract induced apoptosis through caspase-3 activation in both human malignant melanoma (SK-Mel-28) and human cervical cancer (HeLa) cell lines. The cytotoxic activity relies on the presence of the neolignans (balanophonin), and phenol derivatives (gallic acid), showing a pro-apoptotic effect on the tested cell lines, and the neolignan, cedrusin, with a cytotoxic effect on A375 and HeLa cells.

Alginate beads as a carrier for omeprazole/SBA-15 inclusion compound: A step towards the development of personalized paediatric dosage forms.

The treatment of gastro-esophageal reflux disease (GERD) shows several issues among paediatric patients. This work aims to the formulation of enteric alginate beads loaded with omeprazole (OME) allowing age- and weight-related personalized dosages in children. OME was entrapped in SBA-15 mesoporous compound, characterized and loaded into alginate beads by prilling at different OME and alginate concentrations. The beads resulted of homogeneous size, spherical morphology and very consistent in drug loading and distribution. Formulations demonstrated limited swelling and release (about 10%) in simulated gastric fluid (SGF) after 2h and a prolonged release in simulated intestinal fluid (SIF), till 6h, due to a mixed diffusion-case II transport mechanism. The beads were superior to the market product, which showed lower release in SGF but immediate dissolution in SIF. The high alginate beads uniformity and release properties make them a potential novel tool for a personalized treatment of GERD in children.

HRMS Profile of a Hazelnut Skin Proanthocyanidin-rich Fraction with Antioxidant and Anti-Candida albicans Activities.

Roasted hazelnut skins (RHS) represent a byproduct of kernel industrial processing. In this research, a RHS extract (RHS-M) and its fraction RHS-M-F3 enriched in proanthocyanidins (PAs), with antioxidant activity, were characterized in terms of total phenolic compound and PA contents. RHS-M and RHS-M-F3 showed antifungal properties against Candida albicans SC5314 (MIC2 = 3.00 and 0.10 μg/mL and MIC0 = 5.00 and 0.50 μg/mL, respectively), determined by the microbroth dilution method and Candida albicans morphological analysis. No cytotoxic effect on HEKa and HDFa cell lines was exhibited by RHS-M and RHS-M-F3. The metabolite profiling of RHS-M and RHS-M-F3 was performed by thiolysis followed by HPLC-UV-HRMS analysis and a combination of HRMS-FIA and HPLC-HRMS(n). Extract and fraction contain oligomeric PAs (mDP of 7.3 and 6.0, respectively, and DP up to 10) mainly constituted by B-type oligomers of (epi)-catechin. Also, (epi)-gallocatechin and gallate derivatives were identified as monomer units, and A-type PAs were detected as minor compounds.