Flurbiprofen PLGA-PEG nanospheres: role of hydroxy-ß-cyclodextrin on ex vivo human skin permeation and in vivo topical anti-inflammatory efficacy.

In this study, flurbiprofen (FB) loaded poly(d,l-lactide-co-glycolide) (PLGA) and PLGA with poly(ethylene glycol) (PLGA-PEG) nanospheres (NSs) with and without hydroxypropyl-ß-cyclodextrin (HPßCD) were developed as skin controlled delivery systems. X-ray diffraction was used to determine the physical state of the entrapped drug. Results showed that the drug in PLGA NSs was present in the form of a molecular dispersion (dissolved state) in the polymers, whereas in PLGA-PEG NSs, the drug was present in both molecular dispersion and crystalline forms. Furthermore, HPßCD provided solubilization of the free FB present on the surface of the PLGA-PEG NSs and a complete amorphosization of the drug was obtained. Optical analyses using TurbiscanLab(®) demonstrated that HPßCD provided an efficient steric stability of the NSs, preventing particle aggregation. The ex vivo permeation profiles of the NSs and conventional FB solution were evaluated using human skin. Results demonstrated that only PLGA-PEG NSs showed slight permeation improvement. However, after 24h, the FB retained in the skin was about 9-fold higher with NSs compared with the control solution, attributed to the reservoir effect of NSs acting as a depot, sustaining the drug and limiting its systemic absorption. In vivo performance of NSs was evaluated by assessing anti-inflammatory efficacy in TPA-induced mouse ear edema. Topically applied NSs significantly decreased in vivo inflammation compared to the control solution and the anti-inflammatory efficacy of HPßCD NSs was stronger than NSs without HPßCD. In vivo skin irritation evaluated by the in vivo Draize test showed no irritation of the formulations tested.

Role of hydroxypropyl-ß-cyclodextrin on freeze-dried and gamma-irradiated PLGA and PLGA-PEG diblock copolymer nanospheres for ophthalmic flurbiprofen delivery.

Poly(D,L-lactide-co-glycolide) and poly(D,L-lactide-co-glycolide) with poly(ethylene glycol) nanospheres (NSs) incorporating flurbiprofen (FB) were freeze-dried with several cryoprotective agents and sterilized by γ-irradiation. Only when 5.0% (w/v) hydroxypropyl-ß-cyclodextrin (HPßCD) was used, a complete resuspension by manual shaking and almost identical particle size of the NSs was obtained after freeze-drying. In vitro drug release and ex vivo corneal permeation of NSs with and without HPßCD were evaluated. The presence of HPßCD resulted in a reduction of burst effect, providing a more sustained release of the drug. A significant decrease in the FB transcorneal permeation of NSs containing HPßCD was obtained, related to the slower diffusion of FB observed in the in vitro results. The uptake mechanism of the NSs was examined by confocal microscopy, suggesting that NSs penetrate corneal epithelium through a transcellular pathway. Ocular tolerance was assessed in vitro and in vivo by the Eytex™ and Draize test, respectively. Long-term stability studies revealed that γ-irradiated NSs stored as freeze-dried powders maintained their initial characteristics. Stability studies of the resuspended NSs after 3 months of storage in the aqueous form showed that NSs were stable at 4°C, while formulations stored at 25°C and 40°C increased their initial particle size.

Improved and safe transcorneal delivery of flurbiprofen by NLC and NLC-based hydrogels.

Flurbiprofen (FB)-loaded nanostructured lipid carriers (NLCs) based on Compritol®888 ATO (C888; FB-C888NLC) were developed for anti-inflammatory ocular therapy. NLCs prepared by high-pressure homogenization technique following a factorial design had low particle size (<199 nm), high entrapment efficiency (∼90%), and long-term physical stability. Previously optimized NLCs based on stearic acid (SA; FB-SANLC) were prepared for comparison studies. Both formulations were dispersed in freshly prepared carbomer hydrogel (HG) to check the suitability of semisolid-based NLC HGs to enhance the corneal residence time. FB-C888NLC remained in the nanometric range, whereas FB-SANLC suffered an increase in particle size up to 5 µm after incorporation. Consequently, modifications in the crystalline lattice structure were observed for FB-SANLC-enriched HG (HG_FB-SANLC) by X-ray diffractometry. Both HG formulations showed plastic and low or no thixotropic properties, making them suitable for ocular application while maintaining its predominant elastic component as an indicator of good physicochemical stability. Formulations depicted sustained FB release. Ex vivo permeation analysis in isolated rabbit cornea revealed enhanced transcorneal drug permeation from the systems. In vivo ocular tolerance was confirmed by the Draize test. Therefore, NLC are promising and effective systems for ocular delivery of FB.

Interfacial interactions of hydrophobic peptides with lipid bilayers.

Four hydrophobic laminin-related peptides and their corresponding parent peptides were synthesized to use them to target liposomes to tumoral cells. The peptide sequence was YIGSR((NH(2))), and hydrophobic residues linked to the alpha-amino terminal end were decanoyl, myristoyl, stearoyl, and cholesteryl-succinoyl. Before use in biological systems, a physicochemical study was carried out in order to determine their interaction with DPPC bilayers that could compromise both the toxicity and the stability of liposomal preparations. The experiments were based on DSC, fluorescence polarization, outer-membrane destabilization, and vesicle leakage. These peptides showed in general a low interaction with the vesicles, promoting in all cases the rigidification of bilayers. This lack of strong disturbances in the ordered state of phospholipid molecules seems more likely due to the similarity of peptide acyl chains with those of lipids than to the absence of interactions. The bulkiness of cholesteryl derivative as well as its tendency toward aggregation resulted in weak interaction levels except in thermograms. The binding of peptides to the surface of liposomes loaded with doxorubicin resulted in preparations with good entrapment yields and small size, required for long circulating vesicles (especially for the myristoyl derivative). The alternative method based on the reaction of parent peptide to the surface of liposomes through an amide linkage was slightly more efficient when the peptide was linked to the carboxy-terminal end of the DSPE-PEG-COOH-containing liposomes. Nevertheless, the final decision must be made with the simplicity of the procedure and reduction in losses during all the steps of the processes taken into consideration.