A simple HPLC method for the determination of halcinonide in lipid nanoparticles: development, validation, encapsulation efficiency, and in vitro drug permeation

ABSTRACT Halcinonide is a high-potency topical glucocorticoid used for skin inflammation treatments that presents toxic systemic effects. A simple and quick analytical method to quantify the amount of halcinonide encapsulated into lipid nanoparticles, such as polymeric lipid-core nanoparticles and solid lipid nanoparticles, was developed and validated regarding the drug's encapsulation efficiency and in vitro permeation. The development and validation of the analytical method were carried out using the high performance liquid chromatography with the UV detection at 239 nm. The validation parameters were specificity, linearity, precision and accuracy, limits of detection and quantitation, and robustness. The method presented an isocratic flow rate of 1.0 mL.min-1, a mobile phase methanol:water (85:15 v/v), and a retention time of 4.21 min. The method was validated according to international and national regulations. The halcinonide encapsulation efficiency in nanoparticles was greater than 99% and the in vitro drug permeation study showed that less than 9% of the drug permeated through the membrane, indicating a nanoparticle reservoir effect, which can reduce the halcinonide's toxic systemic effects. These studies demonstrated the applicability of the developed and validated analytical method to quantify halcinonide in lipid nanoparticles.

Computationally Prediction of Candidate Agents for Preventing Organ Dysfunction After Brain Death.

BACKGROUND Our aim was to explore the mechanism of post-transplant organ function decrease induced by brain death (BD) and discover a potential candidate drug for improving the survival and organ function after BD. MATERIAL AND METHODS The microarray data developed from the liver tissues after BD were further analyzed by bioinformatics methods. The differentially expressed genes (DEGs) were computationally predicted and the DEGs that involved biological functions were explored by gene ontology (GO) analysis. The candidate agents that could induce the reverse gene signature were predicted based on the Connectivity Map (CMap) database. RESULTS There were total 1374 DEGs, including 589 up-regulated genes and 785 down-regulated genes. Function analysis showed that DEGs were mainly enriched in biological process-related GO terms, such as regulation of transcription, DNA-dependent, inflammatory response, and regulation of phosphorus metabolic process. The down-regulated genes were significantly enriched in transcription factor activity and transcription regulator activity-related molecular function. The down-regulated GO terms exhibited close interaction with each other. CONCLUSIONS The organ function decrease may be attributed by transcription alteration, inflammation response, and metabolic alteration in liver after BD. Spaglumic acid and halcinonide may be potential drugs for preventing organ damage during the BD process.

Clobetasol and Halcinonide Act as Smoothened Agonists to Promote Myelin Gene Expression and RxRγ Receptor Activation.

One of the causes of permanent disability in chronic multiple sclerosis patients is the inability of oligodendrocyte progenitor cells (OPCs) to terminate their maturation program at lesions. To identify key regulators of myelin gene expression acting at the last stages of OPC maturation we developed a drug repositioning strategy based on the mouse immortalized oligodendrocyte (OL) cell line Oli-neu brought to the premyelination stage by stably expressing a key factor regulating the last stages of OL maturation. The Prestwick Chemical Library of 1,200 FDA-approved compound(s) was repositioned at three dosages based on the induction of Myelin Basic Protein (MBP) expression. Drug hits were further validated using dosage-dependent reproducibility tests and biochemical assays. The glucocorticoid class of compounds was the most highly represented and we found that they can be divided in three groups according to their efficacy on MBP up-regulation. Since target identification is crucial before bringing compounds to the clinic, we searched for common targets of the primary screen hits based on their known chemical-target interactomes, and the pathways predicted by top ranking compounds were validated using specific inhibitors. Two of the top ranking compounds, Halcinonide and Clobetasol, act as Smoothened (Smo) agonists to up-regulate myelin gene expression in the Oli-neuM cell line. Further, RxRγ activation is required for MBP expression upon Halcinonide and Clobetasol treatment. These data indicate Clobetasol and Halcinonide as potential promyelinating drugs and also provide a mechanistic understanding of their mode of action in the pathway leading to myelination in OPCs. Furthermore, our classification of glucocorticoids with respect to MBP expression provides important novel insights into their effects in the CNS and a rational criteria for their choice in combinatorial therapies in de-myelinating diseases.

Identification of select glucocorticoids as Smoothened agonists: potential utility for regenerative medicine.

Regenerative medicine holds the promise of replacing damaged tissues largely by stem cell activation. Hedgehog signaling through the plasma membrane receptor Smoothened (Smo) is an important process for regulating stem cell proliferation. The development of Hedgehog-related therapies has been impeded by a lack of US Food and Drug Administration (FDA)-approved Smo agonists. Using a high-content screen with cells expressing Smo receptors and a beta-arrestin2-GFP reporter, we identified four FDA-approved drugs, halcinonide, fluticasone, clobetasol, and fluocinonide, as Smo agonists that activate Hedgehog signaling. These drugs demonstrated an ability to bind Smo, promote Smo internalization, activate Gli, and stimulate the proliferation of primary neuronal precursor cells alone and synergistically in the presence of Sonic Hedgehog protein. Halcinonide, fluticasone, clobetasol, and fluocinonide provide an unprecedented opportunity to develop unique clinical strategies to treat Hedgehog-dependent illnesses.

A qualitative estimate of the influence of halcinonide concentration and urea on the reservoir formation in the stratum corneum.

The existence of a stratum corneum reservoir for topically applied substances is well known. Data concerning the stratum corneum retention time are important for the elaboration of optimal topical treatment. We used the re-occlusion technique followed by skin colour measurements (Chromametry) for the evaluation of the stratum corneum retention time of halcinonide. We found a significant reservoir for halcinonide up to 5 days after the initial application. The retention was found to be corticosteroid concentration and formulation dependent.

[The effect of urea in a topical corticoid preparation on vasoconstriction]. / Einfluss von Urea in einem Kortikoid-Dermatikum auf die Vasokonstriktion.

It was checked by vasoconstriction tests, whether a new topical preparation containing 0.1% halcinonide in combination with 10% urea possibly caused an increase in vasoconstriction. There was no significant difference found between the preparations with and without urea.

[Comparison of 3 corticoid preparations with the vasoconstriction and erythema test]. / Vergleich dreier Kortikoidpräparate mit Vasokonstriktions- und Erythemtest.

The efficacy of a new topic preparation of the corticosteroid Halcinonide containing 5% Urea has been examined and then compared with two commercial preparations of Halcinonide and Betamethasonevalerate respectively. Although these two corticosteroids usually do not belong to the same class of potency, there could not be found any significant difference neither with the vasoconstriction assay nor with the Pyrexal erythema. The influence of the three most important parameters, that are vehicle, occlusion and skin condition, is a subject of discussion.

The influence of topical steroid application on tuberculin skin reactions in healthy persons.

Thirty-seven healthy persons were studied in order to evaluate the influence of topical steroid application on tuberculin skin reactions. Four areas measuring 4 cm in diameter were each treated with 50 micrograms of hydrocortisone cream 1%, or 50% micrograms of halcinonide (Halog) cream 0.1%, or 50 micrograms of unguentum cetacei simplex (cold cream), or not treated. The creams were applied once daily for 3 days before and one day after a tuberculin skin test. After 24 and 48 hours the area of induration were measured. We observed that application of unguentum cetacei simplex increased the size of the induration at the 24-hour reading, but not after 48 hours. Hydrocortisone cream 1% gave the same effect, whereas halcinonide cream (Halog) 0.1% caused ischaemia of the skin and reduced the induration of the skin test after 24 hours, but not after 48 hours. In 12 persons we found that simple rubbing of the skin with halcinonide cream base did not affect the size of the tuberculin skin reaction. In the present study we found that even very potent local steroid application on intact skin could only delay the development of tuberculin skin reactions, but could not diminish their size.