Liposomal Aerosols of Nitric Oxide (NO) Donor as a Long-Acting Substitute for the Ultra-Short-Acting Inhaled NO in the Treatment of PAH.
Liposomal Aerosols of Nitric Oxide (NO) Donor as a Long-Acting Substitute for the Ultra-Short-Acting Inhaled NO in the Treatment of PAH.
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DOI：
10.1007/s11095-016-1911-7
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发表时间：
2016-07
影响因子：
3.7
通讯作者：
Ahsan F
中科院分区：
文献类型：
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作者：
Nahar K;Rashid J;Absar S;Al-Saikhan FI;Ahsan F
本研究旨在开发一种长效一氧化氮（NO）供体二乙烯三胺NONOate（DN）的脂质体制剂，以替代吸入NO（iNO）治疗肺动脉高压（PAH）。通过脂质膜水合方法制备脂质体制剂，并用细胞穿透肽CAR修饰。颗粒的大小，多分散指数（PDI），zeta电位，包封率，储存和雾化稳定性，并在体外释放曲线进行了表征。采用大鼠肺泡巨噬细胞（NR 8383）和转化生长因子β（TGF-β）激活的大鼠肺动脉平滑肌细胞（PASMCs），观察细胞摄取和转运。在离体灌流大鼠肺（IPRL）模型中测定经气管内给药后进入体循环的制剂分数。使用MTT测定和通过测量支气管肺泡灌洗液（BAL）中的损伤标记物来评估制剂的安全性;通过监测野百合碱（MCT）诱导的PAH大鼠模型中平均肺动脉压（mPAP）和体循环压（mSAP）的变化来评价药理学疗效。脂质体大小、zeta电位和包封率分别为171±4 nm、−37± 3 mV和46± 5%，分别脂质体在8 h内释放70±5%的药物，在4°C下储存时稳定。CAR缀合的脂质体被PASMC比没有CAR的脂质体更有效地摄取;大鼠肺泡巨噬细胞对制剂的摄取是最小的。DN-脂质体没有增加肺重量、蛋白质量和BAL液中损伤标志物的水平。肠内CAR-脂质体减少了脂质体从肺进入血液;该制剂在180分钟内使mPAP降低40%。本研究建立了概念验证，即长效NO供体的肽修饰的脂质体制剂可以替代短效iNO。
This study seeks to develop a liposomal formulation of diethylenetriamine NONOate (DN), a long acting nitric oxide (NO) donor, with a goal to replace inhaled NO (iNO) in the treatment of pulmonary arterial hypertension (PAH). Liposomal formulations were prepared by a lipid film hydration method and modified with a cell penetrating peptide, CAR. The particles were characterized for size, polydispersity index (PDI), zeta potential, entrapment efficiency, storage and nebulization stability, and in-vitro release profiles. The cellular uptake and transport were assessed in rat alveolar macrophages (NR8383) and transforming growth factor β (TGF-β) activated rat pulmonary arterial smooth muscle cells (PASMCs). The fraction of the formulation that enters the systemic circulation, after intratracheal administration, was determined in an Isolated Perfused Rat Lung (IPRL) model. The safety of the formulations were assessed using an MTT assay and by measuring injury markers in the bronchoalveolar lavage (BAL) fluid; the pharmacological efficacy was evaluated by monitoring the changes in the mean pulmonary arterial (mPAP) and systemic pressure (mSAP) in a monocrotaline (MCT) induced-PAH rat model Liposome size, zeta potential, and entrapment efficiency were 171±4 nm, −37±3mV, and 46±5%, respectively. The liposomes released 70±5% of the drug in 8 h and were stable when stored at 4°C. CAR-conjugated-liposomes were taken up more efficiently by PASMCs than liposomes-without-CAR; the uptake of the formulations by rat alveolar macrophages was minimal. DN-liposomes did not increase lung weight, protein quantity, and levels of injury markers in the BAL fluid. Intratracheal CAR-liposomes reduced the entry of liposomes from the lung to blood; the formulations produced a 40% reduction in mPAP for 180 minutes. This study establishes the proof-of-concept that peptide modified liposomal formulations of long-acting NO donor can be an alternative to short-acting iNO.