Keum Taekwang, Noh Gyubin, Seo Jo-Eun, Bashyal Santosh, Lee Sangkil
College of Pharmacy, Keimyung University, 1095 Dalgubeol-daero, Dalseo-gu, Daegu 42601, Korea.
Center for Forensic Pharmaceutical Science, 1095 Dalgubeol-daero, Dalseo-gu, Daegu 42601, Korea.
Pharmaceuticals (Basel). 2020 Nov 21;13(11):408. doi: 10.3390/ph13110408.
Buccal tissues are considered one of the potential alternative delivery route because of fast drug absorption and onset of action due to high vascularization and a non-keratinized epithelial membrane. In this study, the effect of Penetratin on the permeation of salmon calcitonin (sCT), a model macromolecular peptide drug, through TR146 buccal cells and porcine buccal tissues has been evaluated. To observe permeation profile of sCT, TR146 buccal cells were treated with Alexa 647 conjugated sCT (Alexa 647-sCT) with different concentrations of fluorescein isothiocyanate -labeled Penetratin (FITC-Penetratin) ranging from 0 to 40 μM, and analyzed using flow cytometry and confocal laser scanning microscopy. Intracellular penetration of FITC-Penetratin rapidly increased at low concentrations from 0 to 15 μM and it gradually increased at concentrations above 15 μM. Intracellular penetration of Alexa 647-sCT enhanced with the increase of FITC-Penetratin concentration. When TR146 cell layers and buccal tissues were co-treated with sCT and Penetratin as permeation enhancer, the flux of sCT increased as per Penetratin concentration. Compared to the control, 12.2 μM of Penetratin enhanced the flux of sCT in TR146 cell layers and buccal tissues by 5.5-fold and 93.7-fold, respectively. These results strongly suggest that Penetratin may successfully act as a non-invasive permeation enhancer for macromolecular peptide drug delivery through buccal routes.
由于颊组织血管丰富且上皮膜无角化，药物吸收快、起效迅速，因此被认为是一种潜在的替代给药途径。在本研究中，评估了穿膜肽对鲑鱼降钙素（sCT，一种大分子肽类药物模型）透过TR146颊细胞和猪颊组织的渗透作用。为观察sCT的渗透情况，用不同浓度（0至40μM）的异硫氰酸荧光素标记的穿膜肽（FITC -穿膜肽）处理TR146颊细胞与Alexa 647偶联的sCT（Alexa 647 - sCT），并采用流式细胞术和共聚焦激光扫描显微镜进行分析。FITC -穿膜肽在低浓度（0至15μM）时细胞内渗透迅速增加，在浓度高于15μM时逐渐增加。Alexa 647 - sCT的细胞内渗透随着FITC -穿膜肽浓度的增加而增强。当TR146细胞层和颊组织与sCT和作为渗透促进剂的穿膜肽共同处理时，sCT的通量随穿膜肽浓度增加而增加。与对照组相比，12.2μM的穿膜肽使TR146细胞层和颊组织中sCT的通量分别提高了5.5倍和93.7倍。这些结果有力地表明，穿膜肽可能成功地作为一种非侵入性渗透促进剂，用于通过颊途径递送大分子肽类药物。
