Solubility and Permeability Enhancement of Oleanolic Acid by Solid Dispersion in Poloxamers and γ-CD.

Oleanolic acid (OA) is a pentacyclic triterpenoid widely found in the Oleaceae family, and it represents 3.5% of the dry weight of olive leaves. OA has many pharmacological activities, such as hepatoprotection, anti-inflammatory, anti-oxidant, anti-diabetic, anti-tumor, and anti-microbic activities. Its therapeutic application is limited by its poor water solubility, bioavailability, and permeability. In this study, solid dispersions (SDs) were developed to overcome these OA limitations. Solubility studies were conducted to evaluate different hydrophilic polymers, drug-to-polymer ratios, and preparation methods. Poloxamer 188, Poloxamer 407, and γ-CD exhibited the highest increases in terms of OA solubility, regardless of the method of preparation. Binary systems were characterized using differential scanning calorimetry (DSC), X-ray diffraction (XRPD), and Fourier transform infrared spectroscopy (FTIR). In addition, pure compounds and SDs were analyzed using scanning electron microscopy (SEM) in order to observe both the morphology and the particle surface. In vitro dissolution studies were performed for P407, P188, and γ-CD SDs. Preparation using the solvent evaporation method (SEM) produced the highest increase in the dissolution profiles of all three polymers with respect to the OA solution. Finally, the effect of SDs on OA permeability was evaluated with an in vitro parallel artificial membrane permeability assay (PAMPA). The formulation improved passive permeation across the simulated barrier due to OA increased solubility. The dissolution and PAMPA results indicate that the amorphization of OA by SD preparation could be a useful method to enhance its oral absorption, and it is also applicable on an industrial scale.

Evaluation of the increase of the thymoquinone permeability formulated in polymeric micelles: In vitro test and in vivo toxicity assessment in Zebrafish embryos.

Thymoquinone (TQ) is a natural compound present in the essential oil and in the fixed oil of Nigella sativa L. Like many natural substances, it is characterized by poor aqueous solubility and low stability which limit its bioavailability. Soluplus®-Solutol® HS15 polymeric micelles (TQ-MP) were developed to increase the permeability of TQ with particular attention to overcoming intestinal barrier and the blood brain barrier, for possible oral and parenteral administration. The optimized micelles have dimensions < 100 nm and PdI < 0.2 indicating that the formulation was homogeneous as confirmed also by TEM experiments. EE% was 92.4 ± 0.3%. Stability studies showed a stable formulation following subsequent dilutions and in the gastric-intestinal media. In vitro studies have revealed that the carrier enhances the permeability of TQ in the intestine and in the blood-brain barrier using Parallel Artificial Membrane Permeability Assay (PAMPA) assay and cellular tests with Caco-2 cells and hCMEC/D3 monolayer cells. Up-take study, cell viability and cytotoxicity studies were also conducted. Fluorescent micelles (FITC-MP), were also optimized to perform in vitro up-take study in Caco-2 cells and to study their toxicity in Zebrafish model. The toxicity was evaluated on three lines of Zebrafish: wild type, transgenic line Tg(Myl7:EGFP) in which cardiomyocytes are marked with green fluorescence protein and Tg(flk1-GFP) line which expresses GFP under the control of the vascular endothelial growth factor receptor 2 (vegfr2) promoter.