Cancer is one of the leading causes of death in the world and protein therapeutics play an important role in combating this disease. Novel nanocarriers are needed for optimal delivery, enhanced therapeutic effect, and protection of proteins. Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles are commonly used, since they are non-toxic, biodegradable, and allow sustained release of the active pharmaceutical ingredient (API). Accurate quantification of the therapeutic inside these nanocarriers is essential for further development and precise in vivo experiments, especially for determining correct therapeutic dose. Bicinchoninic acid (BCA) assay is one of the most popular methods of protein quantification, known for its low sensitivity to common surfactants. However, large discrepancies between published results are often observed, with determined protein encapsulation efficiencies (EE) varying from 20 to 80%. We investigate the interference of excipients or the combination of excipients, on accurate EE determination of two different PLGA nanoparticle formulations using the microBCA assay. The EE was determined using multiple methods: by measuring the un-encapsulated protein (indirect approach) and directly by extracting the protein using sodium hydroxide and dimethyl sulfoxide extraction. We show significant differences between the methods, highlight the most common pitfalls, and show the importance of using correct standards in assessing EE.

High-throughput emulsion electrospinning is a technology that can enable practical nanofiber application for drug delivery. Core-shell structure of the electrospun fibers allows the encapsulation of the active pharmaceutical ingredients (APIs), protects their activity, and controls their release rate. However, electrospinning using high flow rates usually requires high electric fields that may negatively affect the activity of the biomolecules. Moreover charged APIs tend to migrate to the surface of the fibers during the electrospinning process leading to the high burst release. That is disadvantageous when long-term sustained release is needed. We have investigated the influence of the electrospinning parameters such as distances between the electrode and collector and the applied voltages to both, activity of the encapsulated proteins and their burst release. We have also tested the influence of number of the stabilizers, e.g. trehalose, pluronic, polyvinylpyrrolidone, on their ability to preserve the protein activity, and the influence of the different molecular weights of polyvinyl alcohol on the ability to sustain the release. Our results demonstrate the importance of the water phase composition to both activity and release and are critical for further understanding of the processes taking place during the emulsion electrospinning.

The determination of the in vitro release profile is a useful tool during product development, since it can provide essential data on semisolid product microstructure. The present work aimed at providing the assumptions to assist a sustainable improvement of an anti-inflammatory semisolid dosage form pharmaceutical performance. The qualitative/quantitative composition and the production process were already well-established, however, there were some parameters that lacked optimization, namely: (i) sodium hydroxide content (ii) carbopol viscosity and finally (iii) mixing time. Since these parameters induced fluctuations in the microstructure profile, they were identified as critical. Efforts were then made to rationalize, predict and ultimately maximize the effects of these parameters on the product pharmaceutical performance. For that, a 2³ full factorial design was employed to assess the impact of the above mentioned parameters, on the pharmaceutical performance. During this optimization phase, all manufactured batches were produced at a laboratory scale.

Our results enabled the identification of three main formulation groups: high (659.53-723.67 µg/cm²/√h), moderate (598.99-651.14 µg/cm²/√h) and low release rate formulations (499.30-507.59 µg/cm²/√h). These results were noteworthy correlated with sodium hydroxide content (higher sodium hydroxide inducing higher release), and on a smaller degree to the carbopol viscosity (higher viscosity yielding superior drug release).

To confirm these assumptions, the obtained outcomes were translated from lab to industrial scale, with the sole aim to validate the working conditions in line with the predefined quality target product profile (QTPP). As widely known, semisolid microstructure is highly dependent on batch production size. The same trends were observed, even though IVRR suffered a slight decrease when compared to the lab scale batches. Nevertheless, quality target and response constraints monitoring during DoE laboratory studies, enabled a successful implementation of the optimal production conditions at an industrial scale. By doing so, a product with superior pharmaceutical performance was attained.

The present investigation was carried out to develop taste-masked orally disintegrating tablet containing Dexketoprofen for evaluating the effect of the coating amount on product’s quality attributes via Quality by Design (QbD) systematically roadmap.

Dexketoprofen, S(+)-enantiomer of bitter taste ketoprofen, involves in arylalkil group which is the most frequently used analgesic in the management of acute and chronic pain. Bitter-taste active pharmacological ingredients should involve taste masking approach. For this purpose, the bitter taste dexketoprofen particles were coated pH-dependent methacrylates polymer in which one of the methods of taste-masking as a taste-masking agent.

The experimental design was enforced with four-factor three-level Box-Behnken method within the framework of response surface modeling (RSM). Ready to use matrix excipient, Eudragit RS 30D, dextrates, aroma, tablet pressing force were chosen as independent factors, and were assessed on four dependent factors dissolution rate, disintegration time, tablet hardness, and friability.

Our findings indicate that when tablet pressing force is applied as 250 PSI, the tablets disintegrate below 1 minute and friability value's is under 1%. Disintegration time increases as the coating amount increases. However the pareto charts shows engrossingly that the dissolution rate is affected mostly by tablet pressing force in first, third, fifth time points, and by matrix excipient and coating in tenth, fifteenth, twentieth, thirtieth time points.

It was concluded that QbD study helped to understand hoe coating amount and process variables impacting the dissolution rate, disintegration time, tablet hardness and friability of Dexketoprofen orally disintegrating tablet (ODT) finished product.

Optimization of transdermal formulations requires solving simultaneous challenges as the selection of release polymers. The interactions between the formulation components must be taken as a way to modulate its performance. Selection of acrylic polymers with different functionalizations for the transdermal formulation of a tertiary amine drug (ropinirole HCl) have been investigated. Aim of this work is to characterize the influence over drug release of certain experimental interactions. Solubility-crystalization and pharmacopoeial release tests have been used to evaluate the influence of drug loading and the pH of the release media. Area under the curve of dissolved amounts and percentage of release have been used as discriminant variables in mutual influence with the physical state of the drug. Elucidation of release mechanisms has been performed with data fitting of relevant modelystic equations. Fickian release and erosion contribution have been related with drug loading and the risk of burst effects. In conclusion, a rationale to select the best suitable polymer for ropinirole HCl has been demonstrated in terms of efficiency and extent of release.

Osteoarthritis is frequently treated in veterinary with non-steroidal anti-inflammatory drugs such as carprofen (CP). The enhancement of its action over the articular cartilage can be achieved by increasing drug uptake into the cartilage, alongside its site of action, and anticipating its rapid distribution towards bloodstream. A pharmacokinetic study to evaluate carprofen nanoparticles (NP) after intraarticular administration (IA) in rabbits has been performed through a modeling allometric approach. The pharmacokinetic analysis of plasma profiles showed a rapid CP distribution outwards synovial chamber but mainly remaining in plasma (Vc = 0.14 L/5 Kg), according to its high protein-binding. The absorption data modeling showed the occurrence of two different release-absorption rate processes after nanoparticles administration in the synovial space, i.e., a fast rate process causing a burst effect and involving the 59.5% of the total CP absorbed amount and a slow rate process, involving the 40.5%. Interestingly, the CP burst effect inside the joint space enhances its diffusion towards cartilage resulting in CP accumulation about three times higher concentrations than in plasma. In line with these results, the normalised-by-dose AUC values after IA administration were 80% lower than those observed after the intravenous. Moreover, the slower slope of the concentration-time terminal phase after IA administration vs. IV suggested a flip-flop phenomenon (0.35 h-1 vs. 0.19 h-1). Of note, CP clearances are predictive of the PK profile of CP in healthy humans (0.14 L/h/5 Kg vs. 2.92 L/h/70 Kg) although an over-estimation has been detected for cats or dogs (10 times and 4 times respectively). This fact could probably be attributed to inter-species metabolic differences. In summary, despite the limited number of animals used, this study shows that the rabbit model could be suitable for a predictive evaluation of the release enhancement of CP-NP towards the biophase in arthritic diseases not due to sterical retention of the formulation.

Many neuroactive drugs are characterized by poor solubility, hampering their therapeutic potential and clinical research studies. For instance, the lipophilic molecules dimethylfumarate, retinyl palmitate, progesterone and URB597 can be employed in the treatment of relapsing remitting multiple, early brain injury, learning deficits and/or traumatic brain injuries. In this study the possibility to encapsulate these drugs in lipid nanoparticles is investigated. Solid lipid nanoparticles and nanostructured lipid carriers have been produced by melt and ultrasonication of stearic triglyceride or a mixture of stearic triglyceride and caprylic/capric triglycerides. Mean diameters and morphology of lipid particles were studied by photon correlation spectroscopy, cryo-transmission electron microscopy and x-ray diffraction, while encapsulation efficiency and in vitro drug release have been determined by HPLC. A behavioural study was conducted in rats to study the capability of lipid nanoparticles containing URB597 to alter behaviours relevant to psychiatric disorders after intranasal administration. At this regard the nanoparticle surface has been modified by polysorbate 80, in order to obtain “stealth” nanoparticles. The nanoencapsulation strategy allowed to increase drug solubility with respect to unphysiological solvent or solvent mixtures usually employed for animal and clinical studies. In particular, retinyl palmitate solubility in nanostructured lipid carriers has been increased up to 8-fold. Moreover rat behavioral effects observed by nanoencapsulated URB597 administered intranasally suggest the therapeutic potential of this non-invasive route to treat social dysfunctions, such as autism.

Certain challenges like the presence of highly complex structure (blood-brain barrier (BBB)), P-glycoprotein efflux, and the particular enzymatic activity stand in the way of the successful delivery of the drug moieties to the brain and make them fruitless. Many efforts have been conducted to overcome the previous. Direct delivery of drugs to the brain after the intranasal application is one of those strategies since it holds a great hope to raise the chances of drug moieties to the brain. Nanoparticles could be the potential to improve nose-to-brain drug delivery since they are able to protect the encapsulated drugs from biological and/or chemical degradation and increase their penetration across biological barriers. Based on the fact that neuroinflammation is associated with neuron death and neurodegenerative diseases like Alzheimer’s, nonsteroidal anti-inflammatory drugs (NSAIDs) might play a positive role in the disease. The present study aimed to employ the QbD approach for the first time in optimizing polymeric and lipid-based nanoparticles for the nose-to-brain delivery of Meloxicam (MEL), and to perform a comparison between the pure drug and the formulated nanosystems regarding dissolution profiles, permeability, and mucoadhesiveness.

The mouth can be affected by important inflammatory processes resulting from localized or systemic diseases such as diabetes, AIDS and leukemia among others, and are manifested in various types of buccal sores typically presenting pain [1]. The present work focuses on the design, formulation, and characterization of four semi-solid formulations for oral mucosa in order to symptomatically treat these painful processes. The formulations have two active pharmaceutical ingredients, triamcinolone acetonide (TA) and lidocaine hydrochloride (LIDO). The formula also contains Orabase^® as an excipient which is a protective, hydrophobic, and anhydrous adhesive vehicle, used to retain or facilitate the application of active pharmaceutical ingredients (API) to the oral mucosa. After designing the formulations, the validation of the analytical method was performed to achieve reliable analytical results. Franz-type diffusion cells were used to perform drug release studies using synthetic membrane, and permeation studies using buccal mucosa, permitting the estimation of the amount and rate of TA permeated across this mucous membrane. As well, the amount of TA retained within the tissue was estimated, where it performs its anti-inflammatory activity, and showing no significant differences between the 0.05% TA + LIDO and 0.1% TA + LIDO formulations (p > 0.05). Therefore, results evidence the suitability of the administration of the lowest concentration of TA tested, achieving similar efficacy, and decreasing the potential systemic effects of corticoid administration. Besides, sublingual permeation studies were carried out to evaluate a scenario of a continuous contact of the tongue with the applied formulation. The four formulations studied show a pseudoplastic and thixotropic behaviour, ideal for topical application. These results evidence the potential of these topical formulations for the treatment of inflammatory processes in the buccal mucosa.

The past few years have witnessed major developments in nanotechnology with great potential in powering new therapeutic tools for cancer management. Our goal in this study was to develop a biocompatible nanoshuttle for the efficient delivery of 5FU in colorectal cancer patients. Silk fibroin/Poly(ethylene glycol) nanoparticles (SF/PEG NPs) were obtained and further loaded with 5FU. These nanoshuttles were characterized in terms of: morphological properties, size and size-distribution, drug uptake and release potential as well as in vitro cytotoxicity potential screening. The SF/PEG + 5FU NPs cytotoxicity was determined on HT-29 cells after determination of the lethal dose 50 and targeted the evaluation of the cells viability, proliferation potential and migration and invasion potential. The inflammatory profile of RAW 264.7 macrophage cells was also detrmined by flowcytometry. The basic cytotoxicity screening revealed that the pristine SF/PEG NPs displayed a good biocompatibility while the 5FU loaded NPs induced cytotoxic effects on HT-29 cells. More, the 5FU loaded SF/PEG NPs significantly reduced the migration and invasion processes as compared with the unloaded NPs. Lastly, we observed that the cytokines inflammatory profile was significantly altered after the treatment with the 5FU loaded SF/PEG NPs as compared with the unloaded nanoshuttles.