Preparation, Physicochemical Characterisation and DoE Optimisation of a Spray-Dried Dry Emulsion Platform for Delivery of a Poorly Soluble Drug, Simvastatin.

In the presented study, insight into the development and optimisation of the dry emulsion formulation and spray drying process is provided. The aim was to facilitate the dissolution of the poorly soluble, highly lipophilic drug, simvastatin, by forming spray-dried dry emulsion particles having adequate powder flow properties, while assuring sufficient drug content. Simvastatin and a mixture of caprylic, capric triglyceride and 1-oleoyl-rac-glycerol were employed as a model drug and solubilising oils, respectively. A matrix of the dry emulsions was composed at a fixed ratio mixture of mannitol and HPMC. Tween 20 was used in low amounts as the primary emulsion stabiliser. To facilitate process optimisation, a DoE surface response design was used to study the influence of formulation and process parameters on the particle size distribution, powder bulk properties, emulsion reconstitution ability, drug stability and process yield of spray-dried products. Two-fluid nozzle geometry was identified, studied and confirmed to be important for most product critical quality attributes. Models obtained after the study showed acceptable coefficients of determination and provided good insight in the relationship governing the process and product characteristics. Five model optimised products showed adequate process yield, suitable particle size distribution, good reconstitution ability and improved dissolution profile, when compared to a non-lipid-based tablet and the pure drug. However, the obtained dry emulsion powders exhibited poor flow character according to the Carr index. The optimised product was further analysed with NMR during lipolysis to gain insight into the species formed during digestion and the kinetics of their formation.

Rational design to biologics development: The polysorbates point of view.

Formulation development is an essential part of any biopharmaceuticals development programme, and this will affect quality, safety and efficacy of the final drug product. The vast majority of biopharmaceuticals on the market are therapeutic proteins; however, these are less stable compared to conventional pharmaceuticals. To counter aggregation, denaturation and surface adsorption of proteins in solution, surfactants are added to the formulations; however, the choice of the best formulation is a challenge that is faced during formulation development. Polysorbates are the most widely used surfactants in the pharmaceutical industry and are presented in >80% of commercial monoclonal antibody formulations. In this review, we provide a general overview of polysorbates and their issues, and the characteristics that have to be taken into account during formulation development. Degradation of polysorbates, namely by hydrolysis and/or oxidation, is one of the main concerns associated with their use. Furthermore, degradation of polysorbates is determined by formulation composition, pH and storage conditions, therefore underlining the importance and complexity of protein formulation development using polysorbates. A need-based approach should be used for correct selection of excipients in protein formulations that contain polysorbates.

Comparison of natural and synthetic surfactants at forming and stabilizing nanoemulsions: Tea saponin, Quillaja saponin, and Tween 80.

HYPOTHESIS: This study compared the interfacial and emulsification properties of tea saponins, quillaja saponins, and Tween 80. We hypothesized that tea saponins are an effective and sustainable source of plant-based emulsifiers that could replace synthetic or animal-based emulsifiers in many commercial applications. EXPERIMENTS: Interfacial tension measurements were used to characterize the behavior of the three surfactants at an oil-water interface. The emulsifying properties of the surfactants were determined by preparing oil-in-water emulsions containing 10 wt% medium chain triglycerides (MCT) and varying surfactant levels (0.1-2 wt%) using high-pressure homogenization (pH 7). The impact of surfactant type on emulsion formation and stability was determined by measuring particle size, zeta-potential, microstructure, and creaming stability. FINDINGS: The tea saponins were capable of producing nano-scale droplets (d32 < 200 nm) at low surfactant-to-oil ratios (SOR < 0.1). Emulsions containing tea saponins remained stable to droplet aggregation when exposed to various temperatures (30-90 °C), salt levels (0-200 mM NaCl), and pH values (3-9). However, droplet flocculation and/or coalescence occurred under highly acidic (pH 2) and high ionic strength (300-500 mM NaCl) conditions. Tea saponin-coated oil droplets appeared to be mainly stabilized by a combination of electrostatic and steric repulsion. The tea saponins behaved similarly or better than the other two emulsifiers under most conditions. These results suggest that tea saponins are effective plant-based surfactants that may have applications in commercial products.

The effect of formulative parameters on the size and physical stability of SLN based on "green" components.

Cocoa butter (CB) is a largely used excipient in pharmaceutical field. Aim of this work was to set formulative parameters for the preparation of SLN based on "green" lipid matrix for drug delivery as natural, both human and environmental safe systems. Double emulsion technique (w1/o/w2) was selected for SLN preparation. The effect on the dimensional properties of different surfactants (Tween 80 and PEG 40 monostearate) and co-surfactants (PEG400 monostearate, Emulium® Kappa2 and Plurol®Stearique) at different concentrations was evaluated. Stability tests were performed. SLN dispersions were exsiccated and the effect of the dried process on SLN size was evaluated. The influence of temperature on SLN dimensions was investigated at 37 °C. MTT test was performed on raw materials and formulations. The w1/o/w2 is suitable, rapid and economic technique for the preparation of CB SLN. Tween 80-Plurol Stearique combination gives the best results: particles size less than 400 nm and PI of about 0.4 are obtained when PS 2% is used. Both raw materials and formulations are safe. The importance to evaluate the effect of different surfactant and/or co-surfactant on the dimensional properties of SLN is evident by selecting substances with preferable safety profiles, and favorable environmental properties to develop stable "green" SLN.

In Vitro Anti-inflammatory and Antimicrobial Activities of Azithromycin After Loaded in Chitosan- and Tween 20-Based Oil-in-Water Macroemulsion for Acne Management.

The objectives of the current investigation are (1) to prepare and characterize (particle size, surface charge (potential zeta), surface morphology by transmission electron microscopy, drug content, and drug release) the azithromycin (AZM, 100 mg)-loaded oil-in-water (o/w) macroemulsion, (2) to assess the toxicity of macroemulsion with or without AZM using RBC lysis test in comparison with AZM in phosphate buffer solution of pH 7.4, (3) to compare the in vitro antimicrobial activity (in Escherichia coli using zone inhibition assay) of AZM-loaded macroemulsion with its aqueous solution, and (4) to assess the in vitro anti-inflammatory effect (using egg albumin denaturation bioassay) of the AZM-loaded macroemulsion in comparison with diclofenac sodium in phosphate buffer solution of pH 7.4. The AZM-loaded macroemulsion possessed the dispersed oil droplets with a mean diameter value of 52.40 ± 1.55 µm. A reversal in the zeta potential value from negative (-2.16 ± 0.75 mV) to positive (+6.52 ± 0.96 mV) was noticed when AZM was added into the macroemulsion. At a 1:5 dilution ratio, 2.06 ± 0.03 mg of drug was released from macroemulsion followed by 1.01 ± 0.01 and 0.25 ± 0.08 mg, respectively, for 1:10 and 1:40 dilution ratios. Antimicrobial activity maintenance and significant reduction of RBC lysis property were noticed for AZM after loaded in the macroemulsion. However, an increment in the absorbance values for emulsion-treated samples in comparison to the control samples was noticed in the anti-inflammatory test. This speculates the potential of the AZM-loaded emulsion to manage inflammatory conditions produced at Acne vulgaris.

Quantitative analysis of oleic acid and three types of polyethers according to the number of hydroxy end groups in Polysorbate 80 by hydrophilic interaction chromatography at critical conditions.

A quantitative characterization of Polysorbate 80 is crucial for its many applications. In this paper we report a quick RP-HPLC method for the quantitative determination of Polysorbate 80. The hydrolysis of Polysorbate 80 to release oleic acid and three types of polyethers was first carried out. A chromatographic method based on liquid chromatography at critical conditions (LCCC) was then developed for an endgroup-based separation of low-molecular-mass polyether. With this method the polyether, irrespective of its molecular-mass, is separated according to endgroups (functionality) due to interactions of the polar endgroups with the hydrophilic stationary phase. The different types of polyethers and oleic acid were identified using on-line electrospray ionization mass spectrometry and quantified by evaporative light scattering detection.

Low molecular weight polyethylenimine-graft-Tween 85 for effective gene delivery: synthesis and in vitro characteristics.

The development of safe and efficient gene delivery systems is still a challenge for successful gene therapy. In this work, low molecular weight polyethylenimine (PEI 2K) was modified by Tween 85, which bears three oleate chains. Tween 85 modified PEI 2K (TP) could condense DNA efficiently, and TP/DNA complexes (TPCs) showed high resistance to salt-induced aggregation and enzymatic degradation. In addition, TP did not show the obvious cytotoxicity. The introduction of Tween 85 led to a significant increase in the cellular uptake of complexes with higher transfection efficiency, which was strongly inhibited by the addition of free Tween 85 in MCF-7/ADR cells, but not in MCF-7 cells. These results indicated that TP could be a potentially safe and effective copolymer for gene delivery, and TPCs could be taken up mainly by Tween 85-mediated endocytosis in MCF-7/ADR cells.

Synthesis of aminated polysorbate 80 for polyplex-mediated gene transfection.

To develop novel gene delivery carriers, aminated polysorbate 80 (P80-NH(2)) was synthesized with strong positively charged properties through the introduction of three amine groups. The resulting P80-NH(2) and DNA polyplex exhibited superb condensation abilities due to the high densities of positively charged amines groups. Size and surface charge of polyplex were shown to be well suited for cellular internalization. In addition, the P80-NH(2) /DNA polyplex demonstrated acceptable transfection efficiency in HeLa cells and was nontoxic relative to the conventional 25-kDa polyethyleneimine system.

Synthesis and properties of methacrylic-functionalized tween monomer networks.

Tween surfactants possess very interesting properties such as biodegradability, biocompatibility, and low toxicity. The synthesis of acrylate monomers by means of the chemical modification of polysorbate surfactants Tween 20, 40, and 60 with unsaturated groups is described. Monomers were obtained as a result of the reaction of methacrylic anhydride with different grades of Tween surfactants. Further polymerization was carried out in tetrahydrofuran, dimethylformamide, and a mixture of water-tetrahydrofuran. Physicochemistry properties of the polymer networks were investigated, and the obtained results reveal that they strongly depend on the type of solvent used during the polymerization, as well as on the concentration of the casting solution. In particular, our study demonstrated that, depending on the solvent boiling point, i.e., the facility to remove the solvent from the polymer matrix, it is possible to predict properties of the network morphology. Moreover, in vitro studies on controlled release were accomplished to demonstrate the possibility of utilizing these new materials as drug delivery systems. All resulting networks represent a novel class of cross-linked polymeric materials useful both in pharmaceutical and chemical applications.

Asymmetrically functionalized, four-armed, poly(ethylene glycol) compounds for construction of chemically functionalizable non-biofouling surfaces.

Asymmetrically functionalized, four-armed, Tween 20 derivatives that formed stable monomolecular films on solid substrates were designed and synthesized. Thiol-modified Tween 20 was used for forming self-assembled monolayers (SAMs) on gold, and maleimide-modified Tween 20 was introduced onto SiO(2) surfaces with SAMs of (3-mercaptopropyl)trimethoxysilane through Michael addition. These structurally modified Tween 20 compounds gave the original characteristics of Tween 20, non-biofouling (from ethylene glycol groups) and functionalizable (from OH groups) properties, to each substrate. The non-biofouling properties of the Tween 20-coated gold and SiO(2) surfaces were investigated by surface plasmon resonance spectroscopy and ellipsometry, and these surfaces showed strong resistance against nonspecific adsorption of proteins. In addition, the biospecific binding of streptavidin was achieved after coupling of (+)-biotinyl-3,6,9-trioxaundecanediamine onto the non-biofouling surfaces through amide-bond formation.

An investigation into the role of surfactants in controlling particle size of polymeric nanocapsules containing penicillin-G in double emulsion.

Preparation, characterization and drug release behavior of loaded polybutyl adipate (PBA) nanocapsules with penicillin-G are described here. The nanocapsules were produced using a double emulsion solvent evaporation technique, using dichloromethane as an organic solvent and Tween and Span as surfactants. In this process, a mixture of glycerin and water was used instead of the traditional stabilizer system in the preparation of double emulsion. The influence of surfactants on the property of nanocapsules was discussed in detail. The effects of Span and Tween to modify the size of the nanocapsules were different. The mean diameters of penicillin-G loaded nanocapsules ranged from 75 nm to 638 nm and were dependent on the types and content of the surfactants. The encapsulation efficiencies and drug release rates were also affected by the surfactants in the preparation process. It was found that the encapsulation efficiencies of penicillin-G enhanced up to 76.8% with the increase in Span and Tween contents. Increasing Span concentration as an inner surfactant results in the remaining of penicillin-G mostly sealed in the inner aqueous phase and increasing Tween concentration as the outer surfactant enhanced the viscosity of external water phase, which decreased the rate of penicillin-G diffusion from the inner water phase to the outer water phase. Interestingly, the in vitro drug release profiles exhibited a significant burst release, followed by a lag phase of little or no release. Penicillin-G loaded nanocapsules with low concentrations of both surfactants tend to have higher burst release. Under optimum formulation conditions, the encapsulation of penicillin-G can reach up to 60% and the burst release can also fall below 45%. In this case, the fact that the nanocapsules have only 130 nm diameter will be important.

Optimization of lipase-catalyzed synthesis of sorbitan acrylate using response surface methodology.

In this study, we have synthesized sorbitan acrylate through response surface methodology, using sorbitan and vinyl acrylate that catalyze immobilized lipase. In order to optimize the enzymatic synthesis of the sorbitan acrylate, we applied response surface techniques to determine the effects of five-level-four-factors and their reciprocal interactions with the biosynthesis of sorbitan acrylate. Our statistical model predicted that the highest conversion yield of sorbitan acrylate would be approx 100%, under the following optimized reaction conditions: a reaction temperature of 40.1 degrees C, a reaction time of 237.4 min, an enzyme concentration of 8%, and a 4.49:1 acyl donor/acceptor molar ratio. Using these optimal conditions in three independent replicates, the conversion yield reached 97.6+/-1.3%.

Enzymatic synthesis of sorbitan methacrylate according to acyl donors.

Recently, sugar polymers have been considered for use as biomaterials in medical applications. These biomaterials are already used extensively in burn dressings, artificial membranes, and contact lenses. In this study, we investigated the optimum conditions under which the enzymatic synthesis of sorbitan methacrylate can be affected using Novozym 435 in t-butanol from sorbitan and several acyl donors (ethyl methacrylate, methyl methacrylate, and vinyl methacrylate). The enzymatic synthesis of sorbitan methacrylate, catalyzed by Novozym 435 in t-butanol, reached an approx 68% conversion yield at 50 g/L of 1,4-sorbitan, 5% (w/v) of enzyme content, and a 1:5 molar ratio of sorbitan to ethyl methacrylate, with a reaction time of 36 h. Using methyl methacrylate as the acyl donor, we achieved a conversion yield of approx 78% at 50 g/L of 1,4-sorbitan, 7% (w/v) of enzyme content, at a 1:5 molar ratio, with a reaction time of 36 h. Sorbitan methacrylate synthesis using vinyl methacrylate as the acyl donor was expected to result in a superior conversion yield at 3% (w/v) of enzyme content, and at a molar ratio greater than 1:2.5. Higher molar ratios of acyl donor resulted in more rapid conversion rates. Vinyl methacrylate can be applied to obtain higher yields than are realized when using ethyl methacrylate or methyl methacrylate as acyl donors in esterification reactions catalyzed by Novozym 435 in organic solvents. Enzyme recycling resulted in a drastic reduction in conversion yields.

Aplicación de una técnica colorimétrica para la evaluación del efecto de preparados de base tensiactiva sobre el estrato córneo / Application of a colorimetric technique for the evaluation of the action of tensiactive based formulations on the stratum corneum

Se propone una técnica basada en una unción con piranina, colorante fluorescente, para explorar y cuantificar alteraciones infraclínicas del estrato cómeo causadas por la aplicación previa y repetida de tensiactivos habitualmente utilizados en formulación de preparados dermatológicos y cosméticos de higiene. La intensidad de la fluorescencia, detectada mediante un programa de análisis de imágenes, es más elevada cuando la eficacia de la barrera hidrolipidica se reduce por la acción de los tensiactivos aplicados. El protocolo estudiado puede resultar de utilidad para la realización de screenings en la fase de preformulación de este tipo de productos (AU)

MF59. Design and evaluation of a safe and potent adjuvant for human vaccines.

MF59 is a safe, practical, and potent adjuvant for use with human vaccines. The formulation is easily manufactured, may be sterilized by filtration, and is both compatible and efficacious with all antigens tested to date. MF59 has been shown to be a potent stimulator of cellular and humoral responses to subunit antigens in both animal models and clinical studies. Toxicology studies in animal models and Phase I-III studies in humans have demonstrated the safety of MF59 with HSV, HIV, and influenza vaccines.