Sobhani Hamideh, Tarighi Parastoo, Ostad Seyed Nasser, Shafaati Alireza, Nafissi-Varcheh Nastaran, Aboofazeli Reza
Department of Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
Iran J Pharm Res. 2018 Summer;17(3):830-850.
This study was planned to explore the capability of nanoemulsions (NEs) consisting of Capryol 90 and oleic acid for the delivery of rapamycin (RAP). Permeability and cytotoxicity of RAP-loaded NEs were also inspected. Pseudo-ternary phase diagrams were created with oleic acid and Capryol 90 (as oil phase) and four surfactants and co-surfactants at various weight ratios (R). Selected NEs from O/W region on the phase diagrams with the drug concentration of 1 mg/mL, were prepared via the spontaneous emulsification technique, characterized for particle size and subjected to stability tests at various temperatures over 9-12 months. Cumulative drug release was determined for a period of 48 h using a dialysis sac. The assay of RAP was determined using HPLC technique. Cytotoxicity of NEs was evaluated by MTT assay on breast cancer cell line, namely SKBR-3. The permeability of RAP-loaded NEs across Caco-2 monolayers was assessed by measurement of TEER (transepithelial electrical resistance) value. The intracellular uptake of coumarin 6-loaded NEs by SKBR-3 cells was also investigated using florescence microscopy. NEs containing oleic acid/Tween 20/propylene glycol, Capryol 90/Tween 20/-propanol, and Capryol 90/Cremophor RH40/Transcutol P showed more cytotoxicity and permeability compared with the RAP methanolic solution. The minimum toxic concentration of RAP in NE formulations was found to be 7.5 µg/mL. The highest intracellular uptake was observed for the NE composed of Capryol 90/Tween 20/-propanol which was in consistent with the results obtained from cytotoxicity and permeability tests. The overall results implicated that this novel carrier was effective for enhancing RAP permeation in Caco-2 cell membrane along with enhancement of cytotoxicity.
本研究旨在探索由辛酸癸酸甘油三酯（Capryol 90）和油酸组成的纳米乳剂（NEs）用于递送雷帕霉素（RAP）的能力。还检测了载有RAP的NEs的渗透性和细胞毒性。用油酸和辛酸癸酸甘油三酯（作为油相）以及四种表面活性剂和助表面活性剂以不同重量比（R）绘制了伪三元相图。从相图上油包水（O/W）区域中选择药物浓度为1 mg/mL的NEs，通过自发乳化技术制备，对其粒径进行表征，并在9至12个月的不同温度下进行稳定性测试。使用透析袋测定48小时内的药物累积释放量。采用高效液相色谱（HPLC）技术测定RAP含量。通过MTT法在乳腺癌细胞系SKBR-3上评估NEs的细胞毒性。通过测量跨上皮电阻（TEER）值评估载有RAP的NEs跨Caco-2单层细胞的渗透性。还使用荧光显微镜研究了SKBR-3细胞对载有香豆素6的NEs的细胞内摄取情况。与RAP甲醇溶液相比，含有油酸/吐温20/丙二醇、辛酸癸酸甘油三酯/吐温20/丙醇以及辛酸癸酸甘油三酯/聚氧乙烯蓖麻油RH40/肉豆蔻酸异丙酯的NEs表现出更高的细胞毒性和渗透性。发现NE制剂中RAP的最低毒性浓度为7.5 µg/mL。观察到由辛酸癸酸甘油三酯/吐温20/丙醇组成的NE具有最高的细胞内摄取量，这与细胞毒性和渗透性测试结果一致。总体结果表明，这种新型载体可有效增强RAP在Caco-2细胞膜中的渗透，并增强细胞毒性。
