Gennari Chiara G M, Franzè Silvia, Pellegrino Sara, Corsini Emanuela, Vistoli Giulio, Montanari Luisa, Minghetti Paola, Cilurzo Francesco
Department of Pharmaceutical Sciences, Università degli Studi di Milano , Via Giuseppe Colombo, 71-20133 Milan, Italy.
Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano , Via Giuseppe Balzaretti, 9-20133 Milan, Italy.
Biomacromolecules. 2016 Jan 11;17(1):46-55. doi: 10.1021/acs.biomac.5b01524. Epub 2015 Dec 14.
This study aimed to identify a new skin penetrating peptide (SPP) able to enhance unfractionated heparin (UFH) permeation through human epidermis by screening a phage display peptide library. The effects of the synthesized heptapeptide (DRTTLTN) on human stratum corneum organization were investigated by ATR-FTIR spectroscopy and molecular dynamics simulation. The DRTTLTN penetration within the human epidermis caused both a fluidization of the stratum corneum lipids and the extension of keratins due to the increase of the contribution of α-helices. The coadministration of DRTTLTN with UFH resulted ineffective in increasing skin penetration due to UFH affinity for keratins. The conjugation of DRTTLTN to UFH by N-(3-(dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride and sodium N-hydroxysulfosuccinimide led to an increase of the flux of 24-36-fold with respect to raw UFH, depending on the adopted synthetic procedure. The new compounds showed a decrease of the antifactor Xa activity of about 4-5 times. DRTTLTN also permitted to increase the fluxes of small model molecules. In conclusion, these data support the use of SPP to enhance the skin penetration of poorly absorbed compounds even in the case of macromolecules as polysaccharides.
本研究旨在通过筛选噬菌体展示肽库，鉴定一种能够增强普通肝素（UFH）透过人表皮渗透的新型皮肤穿透肽（SPP）。通过衰减全反射傅里叶变换红外光谱（ATR-FTIR）和分子动力学模拟研究了合成的七肽（DRTTLTN）对人角质层组织的影响。DRTTLTN在人表皮内的渗透导致角质层脂质的流化以及由于α-螺旋贡献的增加而使角蛋白伸展。由于UFH对角蛋白的亲和力，DRTTLTN与UFH共同给药在增加皮肤渗透方面无效。通过N-（3-（二甲基氨基）丙基）-N'-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚磺酸钠将DRTTLTN与UFH缀合，相对于未处理的UFH，通量增加了24-36倍，这取决于所采用的合成方法。新化合物的抗Xa因子活性降低了约4-5倍。DRTTLTN还能够增加小分子模型分子的通量。总之，这些数据支持使用SPP来增强难吸收化合物的皮肤渗透，即使在多糖等大分子的情况下也是如此。
