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[摘要]
目的 制备西瑞香素纳米混悬剂,并考察其对多种肿瘤细胞增殖的抑制作用。方法 以粒径、Zeta电位、多分散性指数(PDI)为指标,考察稳定剂、药载比、超声温度、均质温度、均质次数对西瑞香素纳米混悬剂反溶剂沉淀法制备的影响,优化最佳处方和工艺条件。观察西瑞香素纳米混悬剂的透射电镜表征,并进行X射线衍射、差示扫描量热分析,考察其在血浆、PBS中的稳定性以及载药量、体外释放情况,采用MTT法比较其对BT474、SKBR-3、A549、HeLa、HepG2细胞的体外细胞毒性。结果 最佳处方和工艺条件:TPGS为稳定剂,药载比1∶1,共同溶解于DMSO中,在超声(25℃,250 W)条件下,缓慢滴注于去离子水中,透析除去有机溶剂,高压均质(25℃,150 MPa)20次,即得西瑞香素纳米混悬剂,其平均粒径为(163.1±5.4)nm,Zeta电位为(-11.4±0.7)mV,PDI为0.15±0.04。西瑞香素纳米混悬剂近乎为球形,大小较均匀;在血浆中稳定存在,不存在溶血现象,满足静脉注射需求,平均载药量为(39.16±1.09)%。西瑞香素纳米混悬剂对5种受试细胞的生长抑制作用显著提高,并呈现剂量相关性,尤其是对SKBR-3、A549、HeLa、HepG2细胞,IC50在2.1~3.4 μg/mL。结论 西瑞香素纳米混悬剂具有小粒径、高载药量、显著肿瘤细胞毒性等优点,在抗肿瘤研究方面具有良好的应用前景。
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[Abstract]
objective To prepare Daphnoretin Nanosuspensions (Dan-NSps) and study its anti-tumor activities in vitro. Methods Dan-NSps were prepared by anti-solvent precipitation method with particle size, Zeta potential, and PDI as indexes, and stabilizer, drug loading ratio, ultrasonic temperature, homogeneous temperature, and homogeneous times were studied to optimize the optimum prescriptions and technological conditions. TEM of Dan-NSps was observed, and X-ray diffraction and differential scanning calorimetry were carried out. Stabilities in plasma and PBS were studied, and the drug loadings and releases in vitro were also compared. MTT assay was used to evaluate the in vitro cytotoxicities against BT474, SKBR-3, A549, HeLa, and HepG2 cells. Results The best preparation method was as following:TPGS was as stabilizer with drug loading ratio 1:1, and was dissolved in DMSO together. Under the condition of ultrasonic (25 W, 250 W), it was slowly dripped into deionized water, and the organic solvent was removed by dialysis. The high pressure homogenization (25℃, 150 MPa) was 20 times. The average particle size of Dan-NSps was (163.1 ±5.4) nm, the Zeta potential was (−11.4 ±0.7) mV, and PDI was 0.15 ±0.04. Naonosuspensions were nearly spherical, and more uniform in size. Dan-NSps remained stable in plasma with no hemolysis, meeting the demand of intravenous administration. The drug loading content was determined to be as high as (39.16 ±1.09)%. MTT assay showed that Dan-NSps had higher cytotoxicities against 5 kinds of cell lines with a dose dependent relationship, especially for SKBR-3, A549, HeLa, and HepG2 cells, which IC50 were at 2.1 to 3.4 g/mL. Conclusion Dan-NSps had advantages of small particle size, high drug load, and significant tumor cell toxicity, which has a good application prospect in the field of anti-tumor research.
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[基金项目]
国家自然科学基金-广东联合基金资助项目(U1401223)