多烯紫杉醇纳米粒的制备、表征及其抗肿瘤作用研究

Study on Preparation, Characterization, and Anti-tumor Activities of Docetaxel Nanoparticles

关键词:  多烯紫杉醇  纳米粒  4T1细胞  药效学

Key words:  docetaxel  nanoparticles  4T1 cell  pharmacodynamics

摘要:

目的 制备多烯紫杉醇（docetaxel，DTX）纳米粒，并进行体内外抗肿瘤研究。方法 采用反溶剂沉淀联合高压均质法制备DTX-G2纳米粒；采用动态光散射法、扫描电镜考察粒径和形态，并对其体外释放、膜毒性、体外抗肿瘤活性进行研究；建立4T1荷瘤小鼠模型，以紫杉醇注射液为对照组，10 mg·kg^-1 iv给药，考察体内抗肿瘤作用。结果 制备的DTX-G2纳米粒粒径为（356.8±6.709）nm，PDI值为（0.147±0.02），Zeta电位为（-14.4±0.07）mV，载药量为（62.3±1.9）%。扫描电镜观察纳米粒为片状。DTX-G2纳米粒体外缓慢释放，在192 h累积释放率达到80.4%；无溶血现象，可采用静脉注射给药；MTT结果显示DTX-G2纳米粒对4T1细胞的毒性强于DTX溶液（IC₅₀，2.374 μg·mL^-1 vs 5.664 μg·mL^-1，P<0.05）；4T1细胞摄取结果显示DTX-G2纳米粒的摄取量显著高于DTX溶液（20.46 vs 11.01，P<0.05）；体内研究中DTX-G2纳米粒对4T1荷瘤小鼠的的抑瘤率显著高于紫杉醇注射液组（75.7% vs 52.4%，P<0.05）。结论 制备的DTX-G2纳米粒载药量高、稳定性好，显著提高了DTX的抗肿瘤效果，有望作为一种新型的药物输送系统应用到DTX的临床治疗中。

Abstract:

OBJECTIVE To prepare docetaxel(DTX) nanoparticles and study its anti-tumor activities in vitro and in vivo.METHODS The DTX-G2 nanoparticles were prepared via the method of anti-solvent precipitation combined high-pressure homogenization. Dynamic light scattering method was used to measure the particle size and scanning electron microscopy was used to observe the morphology. Drug release in vitro, the membrane toxicity and the cytotoxicity were studied. Anti-tumor effect in vivo was investigated on 4T1-bearing mice using Paclitaxel injections as the control at the dose of 10 mg·kg^-1. RESULTS The particle size of DTX-G2 nanoparticles was (356.8±6.709)nm with polydisperse index of 0.147±0.02 and a Zeta potential of (-14.4±0.07)mV, the drug loading capacity was (62.3±1.9)%. The nanoparticles exhibited schistose morphology in SEM. The accumulative release achieved 80.4% in 192 h. DTX nanoparticles presented no hemolytic activity, could be utilized via iv administration. MTT assay displayed that DTX-G2 nanoparticles had stronger cytotoxicity against 4T1 cells than solution (IC₅₀, 2.374 μg·mL^-1 vs 5.664 μg·mL^-1, P<0.05), the uptake ratio of nanoparticles was enhanced significantly(20.46 vs 11.01, P<0.05). The anti-tumor efficacy of DTX was promoted in vivo comparing with Paclitaxel injections (75.7% vs 52.4%, P<0.05). CONCLUSION The DTX-G2 nanoparticles present high drug-loading content, good stability, higher cellular uptake ability, and enhance anti-tumor efficacy which is expected to be a new drug delivery system in clinical trials of DTX.