A Novel Camptothecin Derivative Incorporated in Nano-Carrier Induced Distinguished Improvement in Solubility, Stability and Anti-tumor Activity Both In Vitro and In Vivo
A Novel Camptothecin Derivative Incorporated in Nano-Carrier Induced Distinguished Improvement in Solubility, Stability and Anti-tumor Activity Both In Vitro and In Vivo
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DOI：
10.1007/s11095-008-9795-9
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发表时间：
2009-04-01
影响因子：
3.7
通讯作者：
Gao, Jian-Qing
中科院分区：
文献类型：
--
作者：
Han, Min;He, Cai-Xia;Gao, Jian-Qing
本研究开发了一种油/水纳米乳，以提高新型10-甲氧基-9-硝基喜树碱（MONCPT）的溶解度、稳定性和抗肿瘤活性。以Lipoid E80和cremophor EL为主要乳化剂，通过微流化法制备MONCPT纳米乳。通过动态光散射测量纳米乳液的液滴尺寸。通过膜透析监测体外药物释放。 MONCPT 转化为羧酸盐的动力学在不同 pH 值的磷酸盐缓冲液中进行。 MONCPT纳米乳在兔红细胞中进行溶血。使用尼罗红作为溶剂化显色探针实验了 MONCPT 在纳米乳液中的增溶特性。采用磺胺罗丹明B蛋白染色法测定纳米乳对A549和S180细胞的体外细胞毒性，并在S180荷瘤小鼠中考察其对肿瘤生长的抑制率。采用流式细胞仪分析MONCPT纳米乳对S180细胞的细胞周期影响。还通过荧光图像研究了纳米乳在A549细胞和S180小鼠中的分布。MONCPT以内酯形式掺入纳米乳中，浓度为489μg/ml，比水中的浓度高200倍以上。使用尼罗红作为溶剂化显色探针的实验表明，与离散油滴或连续水相相比，更多的 MONCPT 可能位于纳米乳液的界面表面活性剂层中。纳米乳可以持续释放MONCPT，并且进一步显示出显着推迟MONCPT的水解，在pH 7.4下纳米乳的水解半衰期（11.38 h）比MONCPT溶液（4.03 h）更长。 MOCPT纳米乳对兔红细胞无明显溶血作用。 MONCPT纳米乳通过将细胞阻滞在G2期，使S180细胞和A549细胞的细胞毒活性显着增加，与注射MONCPT诱导的细胞毒活性相比，分别增加23.6倍和28.6倍。与MONCPT纳米乳的体内抗肿瘤活性相关性良好，抑制率为93.6%，而相同剂量下MONCPT注射液的抑制率为24.2%。此外，纳米乳在体外表现出增强的将药物递送至恶性细胞核的能力，并通过体内成像诱导S180荷瘤小鼠肿瘤中的药物积累。制备的纳米乳表现出改善的MONCPT溶解度、稳定性和抗肿瘤活性，为MONCPT的癌症化疗提供了有前景的载体。
An oil/water nanoemulsion was developed in the present study to enhance the solubility, stability and anti-tumor activity of a novel 10-methoxy-9-nitrocamptothecin (MONCPT).MONCPT nanoemulsion was prepared using Lipoid E80 and cremophor EL as main emulsifiers by microfluidization. The droplet size of the nanoemulsion was measured by dynamic light scattering. In vitro drug release was monitored by membrane dialysis. Kinetics of MONCPT transformed into carboxylic salt was performed in phosphate buffer at different pH. Hemolysis of MONCPT nanoemulsion was conducted in rabbit erythrocytes. Solubilization character of MONCPT in nanoemulsion was experimented using Nile red as a solvatochromic probe. In vitro cytotoxicity of the nanoemulsion was measured in A549 and S180 cells using Sulforhodamine B protein stain method, and suppression rate of tumor growth was investigated in S180-bearing mice. The cell cycle effects of MONCPT nanoemulsion on S180 cells were analyzed by flow cytometry. Distribution of the nanoemulsion in A549 cells and S180-bearing mice were also investigated by fluorescence image.MONCPT is incorporated in the nanoemulsion in form of lactone with concentration of 489 A mu g/ml, more than 200 folds higher than that in water. Experiments using Nile red as a solvatochromic probe indicated that more MONCPT might be located in the interfacial surfactant layer of the nanoemulsion than that in discrete oil droplet or continuous aqueous phase. Nanoemulsion could release MONCPT in a sustained way, and it was further shown to notably postpone the hydrolysis of MONCPT with longer hydrolysis half-life time (11.38 h) in nanoemulsion at pH 7.4 than that of MONCPT solution (4.03 h). No obvious hemolysis was caused by MOCPT nanoemulsion in rabbit erythrocytes. MONCPT nanoemulsion showed a marked increase in cytotoxic activity, 23.6 folds and 28.6 folds in S180 cells and A549 cells respectively via arresting the cell at G2 phase, compared to that induced by MONCPT injection. It correlated well to the in vivo anti-tumor activity of MONCPT nanoemulsion with suppression rate of 93.6%, while that of MONCPT injection was only 24.2% at the same dosage. Moreover, nanoemulsion exhibited enhanced capability of delivering drug into malignant cell's nucleus in vitro and induced drug accumulation in tumor in S180-bearing mice using in vivo imaging.The nanoemulsion prepared exhibited an improved MONCPT solubility, stability and anti-tumor activity, providing a promising carrier for cancer chemotherapy using MONCPT.