活性氧/谷胱甘肽双重响应型紫杉醇前药纳米粒的制备与评价

Preparation and Evaluation of Reactive Oxygen Species/Glutathione Dual Responsive Paclitaxel Prodrug Nanoparticle

关键词:  肿瘤微环境  纳米粒  紫杉醇  ROS响应  GSH响应  细胞毒性

Key words:  tumor microenvironment  nanoparticle  paclitaxel  ROS-response  GSH-response  cytotoxicity

摘要:

目的 设计具有活性氧/谷胱甘肽双重响应的紫杉醇前药纳米粒(ProPTX-SS-NPs)，为紫杉醇的应用提供新思路和新方法。方法 以粒径和PDI为指标，考察前药纳米粒的最佳制备方法和工艺；通过电镜观察前药纳米粒的形貌并对其粒径、电位、包封率、载药量等进行考察；考察纳米粒在活性氧和谷胱甘肽环境下的体外释放特性；通过细胞试验考察前药纳米粒的体外细胞毒性和细胞摄取情况。结果 采用最佳工艺制备的纳米粒粒径为(130.20±2.18) nm，分散系数为0.12±0.01，Zeta电位为(–8.45±0.01) mV，载药量为(10.27±1.36)%，包封率为(93.22±2.20)%。前药纳米粒具有良好的活性氧和谷胱甘肽响应特性，并且能够显著抑制MCF-7、HepG2和MDA-MB-231增殖。其对MDA-MB-231细胞的抑制作用最为显著，半数抑制浓度IC₅₀(0.71±0.11)μmol·L^-1，而PTX的IC₅₀为(22.38±3.27)μmol·L^-1。结论ProPTX-SS-NPs具有良好的肿瘤微环境响应性能，具备显著的抗肿瘤活性，是一种极具潜力和应用前景的抗肿瘤纳米系统。

Abstract:

OBJECTIVE To design paclitaxel prodrug nanoparticles with dual reactive oxygen species/glutathione response (ProPTX-SS-NPs), providing new ideas and methods for the application of paclitaxel. METHODS The optimal preparation method and process of prodrug nanoparticles was investigated by using particle size and PDI as indicators; the morphology of prodrug nanoparticles was observed through electron microscopy and their particle size, potential, encapsulation efficiency, drug loading capacity, etc were investigated; the in vitro release characteristics of nanoparticles in reactive oxygen species and glutathione environments were investigated; the in vitro cytotoxicity and cellular uptake of prodrug nanoparticles were investigated through cell experiments. RESULTS The nano particle size prepared by the optimal process was (130.20±2.18)nm, with the PDI of 0.12±0.01, the Zeta potential of (-8.45±0.01)mV, the drug load of (10.27±1.36)%, and the encapsulation rate of (93.22±2.20)%. The prodrug nanoparticles showed reactive oxygen species and glutathione dual responsive release ability, and could significantly inhibit the proliferation of MCF-7, HepG2, and MDA-MB-231. Its inhibitory effect on MDA-MB-231 cells was the most significant. The IC₅₀ of prodrug nanoparticles on MDA-MB-231 cells was (0.71±0.11)μmol·L^-1, while the IC₅₀of PTX was (22.38±3.27)μmol·L^-1. CONCLUSION ProPTX-SS-NPs have excellent tumor microenvironment response performance and significant anti-tumor activity, which is a highly potential and promising anti-tumor nanosystem.