A quantitative UHPLC-MS/MS method for the growth hormone-releasing peptide-6 determination in complex biological matrices and transdermal formulations.

Growth hormone-releasing peptide-6 (GHRP-6) is part of a group of small synthetic peptides with potent GH-releasing activity that have gained attention in the last two decades by virtue of their cyto- and cardioprotective effects. Despite numerous preclinical studies highlighting the potential cardiovascular benefits of GHRP-6, confirmation of clinical efficacy is still awaited. Recent advances in transdermal drug delivery systems have been made to address challenges related to the poor skin permeation rate of peptides by using pain-free microneedle (MN) devices. Accordingly, highly sensitive and validated analytical methods are required for the potential clinical translation of MN-based peptides. The ultra-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) methods developed in this study aimed to quantify GHRP-6 in biological matrices (plasma, skin) and dissolving polymeric MNs. UHPLC/MS-MS method detection limits of 0.1, 1.1, 0.9 and 1.5 ng/mL were achieved in neat solution, plasma, MN polymer solution, and skin matrices, respectively. Method validation also involved assessment of precision, accuracy, limits of quantification, linearity of matched calibration curves (R2 > 0.990), extraction recovery, matrix effect, stability studies, selectivity, and carry-over effect. Additionally, quality control samples were analyzed at three concentration levels to determine recovery (85-109%) and accuracy/bias (3.2-14.7%). Intra- and inter-day precision were within the range of acceptance (RSDs of 3.0-13.9% and 0.4-14.5%, respectively). The validity and applicability of such methods were successfully demonstrated for transdermal GHRP-6 delivery using GHRP-6-loaded MN patches applied to pig skin.

Preparation, Characterization and Evaluation of Organogel-Based Lipstick Formulations: Application in Cosmetics.

1,3:2,4-Dibenzylidene-D-sorbitol (DBS) and 12-hydroxystearic acid (12-HSA) are well-known as low-molecular-weight organogelators (LMOGs) capable of gelling an organic liquid phase. Considering their unique chemical and physical properties, we assessed their potential effects in new lipstick formulations by discrimination testing; in vitro measurements of the sun protection factor (SPF); and thermal, mechanical and texture analyzes. DBS and 12-HSA were used to formulate four types of lipsticks: L1 (1% DBS), L2 (10% 12-HSA), L3 (1.5% DBS) and L4 (control, no LMOGs). The lipsticks were tested for sensory perception with an untrained panel of 16 consumers. LMOG formulations exhibited higher UVA protection factor (UVA-PF) and in vitro SPF, particularly in the 12-HSA-based lipstick. Regarding thermal properties, the 12-HSA-based lipstick and those without LMOGs were more heat-amenable compared to thermoresistant DBS-based lipsticks. The results also showed the viscoelastic and thermally reversible properties of LMOGs and their effect of increasing pay-off values. In general, the texture analysis indicated that 12-HSA-based lipstick was significantly harder to bend compared to control, while the other formulations became softer and easier to bend throughout the stability study. This work suggests the potential use of LMOGs as a structuring agent for lipsticks, paving the way towards more photoprotective and sustainable alternatives.

Preparation and characterization of 12-HSA-based organogels as injectable implants for the controlled delivery of hydrophilic and lipophilic therapeutic agents.

Organogels prepared with low molecular weight organogelators to structure liquid oils represent excellent matrices for the controlled delivery of a wide variety of drug molecules. Although studies on organogel systems are reported in the literature, relatively few investigate their potential as gels formed in situ intended for drug delivery. This study reports the development of injectable subcutaneous 12- hydroxystearic acid (12-HSA) organogels for the delivery of both lipophilic and hydrophilic drugs. The rheological characterization (flow, dynamic temperature ramp and amplitude oscillatory measurements) and physicochemical properties (syringeability, swelling and degradation studies), as well as permeability and cytotoxicity were analyzed to gain insights into the influence of the gel composition (surfactant addition, organogelator concentration) on the gelation process and organogel properties. Sol-gel phase transition temperature (Tgel) and gel-sol phase transition temperature (Tmelt) were determined by the tube-inverting method and complementary rheology studies. An increase in 12-HSA concentration led to an augmentation in gel strength and storage (G') and loss (G″) moduli values, evidencing the self-assembly of crystalline gelator structure entrapping the oil phase into a three-dimensional (3D) network. The addition of polysorbate 80 (Tween 80, T80) surfactant molecules in the system caused a weaker gel-like structure, with lower flow rate during syringeability assays, despite its lower apparent viscosity compared to those of 12-HSA organogels. In addition, the swelling studies of 12-HSA/12-HSA T80 organogels as a function of time in phosphate buffered saline (PBS) revealed that the erosion rates were modulated by the organogel compositions. The permeability of acyclovir (ACV) and clotrimazole (CTM), hydrophilic and lipophilic model drugs, respectively, loaded in 12-HSA-based organogels, was assessed in Franz diffusion cells. Organogel-loaded drugs presented lower in vitro release rates and ex vivo drug permeabilities compared to the corresponding drug solutions. Furthermore, 12-HSA T80 organogel could slow down the release of ACV by a factor of about 2.6-fold, up to 6 h, compared to CTM-loaded 12-HSA organogels. Finally, the cytotoxicity of 12-HSA-based organogels was evaluated through in vitro cell viability assays in human foreskin fibroblasts (HFF). Increased 12-HSA concentration resulted in higher cytotoxic effect, with a higher test sensitivity observed for the 3D collagen-embedded cell layer setup matrix versus 2-D cell cultures. Our results support the hypothesis that 12-HSA-based organogels are promising systems for controlled drug delivery as in situ-forming implants.

Atheroprotective and atheroregressive potential of azapeptide derivatives of GHRP-6 as selective CD36 ligands in apolipoprotein E-deficient mice.

BACKGROUND AND AIMS: Scavenger receptor class B member 3, also known as cluster of differentiation-36 (CD36) receptor, is involved in the uptake and accumulation of modified lipoprotein in macrophages, driving atherosclerosis progression. Azapeptide analogs of growth hormone-releasing peptide-6 (GHRP-6) have been developed as selective CD36 ligands and evaluated for their anti-atherosclerotic properties in apoe-/- mice. METHODS: From 4 to 19 weeks of age, male apoe-/- mice were fed a high fat high cholesterol (HFHC) diet, then switched to normal chow and treated daily with 300 nmol/kg of MPE-001 ([aza-Tyr4]-GHRP-6) or MPE-003 ([aza-(N,N-diallylaminobut-2-ynyl)Gly4]-GHRP-6) for 9 weeks. In another protocol, mice were fed a HFHC diet throughout the study. RESULTS: Azapeptides decreased lesion progression in the aortic arch and reduced aortic sinus lesion areas below pre-existing lesions levels in apoe-/- mice which were switched to chow diet. In mice fed a HFHC throughout the study, azapeptides reduced lesion progression in the aortic vessel and sinus. The anti-atherosclerotic effect of azapeptides was associated with a reduced ratio of iNOS+/CD206+ macrophages within lesions, and lowered plasma inflammatory cytokine levels. Monocytes from azapeptide-treated mice showed altered mitochondrial oxygen consumption rates, consistent with an M2-like phenotype. These effects were dependent on CD36, and not observed in apoe-/-cd36-/- mice. CONCLUSIONS: Azapeptides MPE-001 and MPE-003 diminished aortic lesion progression and reduced, below pre-existing levels, lesions in the aortic sinus of atherosclerotic mice. A relative increase of M2-like macrophages was observed in lesions, associated with reduced systemic inflammation. Development of CD36-selective azapeptide ligands merits consideration for treating atherosclerotic disease.

Organogels, promising drug delivery systems: an update of state-of-the-art and recent applications.

Organogels are semi-solid systems with an organic liquid phase immobilized by a three-dimensional network composed of self-assembled, crosslinked or entangled gelator fibers. Organogel applications are various, including chemistry, pharmaceuticals, cosmetics, biotechnologies and food technology. In pharmacology, they are used as drug and vaccine delivery platforms for active ingredients via diverse routes such as transdermal, oral and parenteral. In a close past, their uses as drug delivery systems have been unfortunately hampered by the toxicity of the selected organic solvents. More recently, the synthesis of more biocompatible organogels has strengthened the development of several biomedical and pharmaceutical applications. This review provides a global view of organogels, such as nature, syntheses, characterizations and properties. An emphasis is placed on the most recent technologies used in the design of organogels as potential controlled delivery systems. A particular attention is provided to their newest therapeutic applications.