Novel and Modified Neutrophil Elastase Inhibitor Loaded in Topical Formulations for Psoriasis Management

Abstract

Human neutrophil elastase (HNE) is a serine protease that degrades matrix proteins. An excess of HNE may trigger several pathological conditions, such as psoriasis. In this work, we aimed to synthesize, characterize and formulate new HNE inhibitors with a 4-oxo-β-lactam scaffold with less toxicity, as well as therapeutic index in a psoriasis context. HNE inhibitors with 4-oxo-β-lactam scaffolds were synthesized and characterized by NMR, FTIR, melting point, mass spectrometry and elemental analysis. In vitro cytotoxicity and serine protease assays were performed. The compound with the highest cell viability (AAN-16) was selected to be incorporated in an emulsion (AAN-16 E) and in a microemulsion (AAN-16 ME). Formulations were characterized in terms of organoleptic properties, pH, rheology, droplet size distribution, in vitro drug release and in vivo psoriatic activity. All compounds were successfully synthesized according to analytical methodology, with good yields. Both formulations presented suitable physicochemical properties. AAN-16 E presented the most promising therapeutic effects in a murine model of psoriasis. Overall, new HNE inhibitors were synthesized with high and selective activity and incorporated into topical emulsions with potential to treat psoriasis.

Investigations of Olive Oil Industry By-Products Extracts with Potential Skin Benefits in Topical Formulations

“…The average droplet size for these formulations is higher than that of the Control Cream ( d (0.5) = 16 μm), possibly due to the incorporation of the extracts in the formulations. Additionally, the droplet size results are in accordance with the results obtained in the literature for this type of formulations, which usually ranges from 0.1 to 50 μm [ 42 , 43 , 60 , 61 ].…”

“…The cytotoxicity of the OIBPE was evaluated on the human keratinocyte cell line HaCaT (Cell line Service GmbH, Eppelheim, Germany) and mouse fibroblasts cell line L929 (ATCC ® CCL-1™), using the endpoint MTT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide) reduction method [ 43 , 44 ]. The cells were seeded at 2 × 10 5 cells/well in sterile 96-well culture plates (Greiner, Frickenhausen, Germany), in RPMI 1640 culture medium (Life Technologies, Warrington, UK), supplemented with 10% fetal bovine serum, 100 units of penicillin G (sodium salt) and 100 μg of streptomycin sulfate, and 2 mM L-glutamine (Life Technologies, Warrington, UK), at 37 °C and 5% CO 2 .…”

“…Fluorometric inhibition assays for Human Neutrophil Elastase (HNE, Merck, Darmstadt, Germany) were conducted as previously described [ 43 ], in 0.1 M HEPES pH 7.5 (assay buffer) at 25 °C, in a total volume of 200 μL (containing 20 μL of 0.17 μM HNE in assay buffer, 155 μL of buffer and 5 μL of each extract sample at a final concentration of 0.025 mg/mL).…”

Perspective trends of topical therapy of patients with psoriasis

“…Повышение активности эластазы может, с одной стороны, приводить к разрушению белков клеточного матрикса кожи, с другой -к активации IL-36, тем самым инициируя воспалительный процесс. В пораженной коже больных псориазом установлен высокий уровень экспрессии как нейтрофильной эластазы, так и активированных с ее помощью агонистов IL-36 [12,13].…”

“…Повреждение собственных белков может быть триггером различных иммуноопосредованных заболеваний, в том числе и псориаза. Подавление повышенной активности данного фермента рассматривается как одно из перспективных направлений терапии больных псориазом [12].…”