De Marco Almeida Flavia, Silva Carolina Nunes, de Araujo Lopes Savia Caldeira, Santos Daniel Moreira, Torres Fernanda Silva, Cardoso Felipe Lima, Martinelli Patricia Massara, da Silva Elizabeth Ribeiro, de Lima Maria Elena, Miranda Lucas Antonio Ferreira, Oliveira Monica Cristina
Laboratorio de Tecnologia Farmaceutica, Departamento de Produtos Farmaceuticos, Faculdade de Farmacia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Laboratorio de Venenos e Toxinas Animais, Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Curr Drug Deliv. 2018;15(7):1064-1071. doi: 10.2174/1567201815666180108170206.
PnPP-19 is a 19-amino-acid synthetic peptide previously described as a novel drug for the treatment of erectile dysfunction.
The aim of this work was to evaluate the physicochemical properties of cationic transfersomes containing PnPP-19 and the skin permeation of free PnPP-19 and PnPP-19-loaded transfersomes.
Three different liposomal preparation methods were evaluated. Cationic transfersomes contained egg phosphatidyl choline: stearylamine (9:1 w/w) and Tween 20 (84.6:15.4 lipid:Tween, w/w). Lipid concentration varied from 20 to 40 mM. We evaluated the entrapment percentage, mean diameter, zeta potential and stability at 4 °C of the formulations. The skin permeation assays were performed with abdominal human skin using Franz diffusion cell with 3 cm2 diffusion area at 32 °C and a fluorescent derivative of the peptide, containing 5-TAMRA, bound to PnPP-19 C-terminal region, where an extra lysine was inserted.
Our results showed variable entrapment efficiencies, from 6% to 30%, depending on the preparation method and the lipid concentration used. The reverse phase evaporation method using a total lipid concentration equal to 40 mM led to the best entrapment percentage (30.2 + 4.5%). Free PnPP-19 was able to permeate skin at a rate of 10.8 ng/cm2/h. However, PnPP-19 was specifically hydrolyzed by skin proteases, generating a fragment of 15 amino acid residues. Encapsulated PnPP-19 permeated the skin at a rate of 19.8 ng/cm2/h.
The encapsulation of PnPP-19 in cationic transfersomes protected the peptide from degradation, favoring its topical administration.
PnPP - 19是一种由19个氨基酸组成的合成肽，先前被描述为一种治疗勃起功能障碍的新型药物。
本研究旨在评估含有PnPP - 19的阳离子传递体的物理化学性质，以及游离PnPP - 19和负载PnPP - 19的传递体的皮肤渗透性。
评估了三种不同的脂质体制备方法。阳离子传递体包含蛋黄卵磷脂：硬脂胺（9:1 w/w）和吐温20（脂质:吐温为84.6:15.4，w/w）。脂质浓度在20至40 mM之间变化。我们评估了制剂的包封率、平均直径、zeta电位和4°C下的稳定性。使用Franz扩散池在32°C下对腹部人体皮肤进行皮肤渗透试验，扩散面积为3 cm²，肽的荧光衍生物与PnPP - 19的C末端区域结合，该区域插入了一个额外的赖氨酸，其中含有5 - TAMRA。
我们的结果显示，根据制备方法和所用脂质浓度的不同，包封效率在6%至30%之间变化。使用总脂质浓度等于40 mM的反相蒸发法导致最佳包封率（30.2 + 4.5%）。游离PnPP - 19能够以10.8 ng/cm²/h的速率渗透皮肤。然而，PnPP - 19被皮肤蛋白酶特异性水解，产生一个15个氨基酸残基的片段。包封的PnPP - 19以19.8 ng/cm²/h的速率渗透皮肤。
将PnPP - 19包封在阳离子传递体中可保护该肽不被降解，有利于其局部给药。
