17-AAG聚氰基丙烯酸正丁酯纳米粒的制备及抗肿瘤活性研究

摘要

目的：制备一种新型17-丙烯氨基-17-去甲氧基格尔德霉素聚氰基丙烯酸正丁酯纳米粒(17-allylamino-17-demethoxygeldanamycin poly-butylcyanoacrylate nanoparticles,17-AAG-PBCA-NPs)。方法采用界面聚合法制备17-AAG-PBCA-NPs；采用正交实验筛选最优处方；采用纳米粒度仪、透射电镜、扫描电镜对17-AAG-PBCA-NPs进行表征和鉴定；采用动态透析法测定体外释放药物情况；采用四甲基偶氮唑盐[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]考察17-AAG-PBCA-NPs对5种人肿瘤细胞株增生的抑制作用；以荷S180小鼠为模型,考察17-AAG-PBCA-NPs的抗肿瘤活性。结果界面聚合法制备的17-AAG-PBCA-NPs包封率大于90%,粒径为(180.5±12.0)nm,Zata电位为(-28.38±0.81)mV,表面形态规则均匀；17-AAG-PBCA-NPs呈时间依赖性地抑制肿瘤细胞株增生,动物实验表明,17-AAG-PBCA-NPs抗肿瘤活性比17-AAG强,毒性比17-AAG低。结论采用界面聚合法可制备得到17-AAG-PBCA-NPs,制备方法简单、重现性好、包封率高,并显示了较好的抗肿瘤活性。%Objective To prepare 17-allylamino-17-demethoxygeldana-mycin poly-butylcyanoacrylate nanoparticles ( 17-AAG-PBCA-NPs) and study the anticancer activity of 17-AAG poly-butylcyanoacrylate nanoparticles. Methods 17-AAG was encapsulated in PBCA-NPs by interfacial polymerization method. Using single factor analysis combined with orthogonal design to compare the effect factors on 17-AAG-PBCA-NPs, optimizing the preparation method of 17-AAG-PBCA-NPs. Nanoparticle size analyzer, TEM and SEM were used to identify and analyze the characteristics of 17-AAG-PBCA-NPs. The dynamic dialysis method was used to determine the in vitro release of 17-AAG-PBCA-NPs. Results Optimal dosage of drug mass ratio was 1 : 10. The conditions of the reaction system were:pH 2, 1 200 r/min, F-68, 3% Dextran70. The time of polymerization was 3 hours. The optimal encapsulation efficiency was more than 90%. The morphology of 17-AAG-PBCA-NPs was spherical shape with (180. 5±12. 0) nm in diameter and the Zeta potential ranged from -20 to-30 mV. The release of 17-AAG-PBCA-NPs in vitro was determined by dynamic dialysis method and it has been shown that the release of drug from the PBCA-NPs exhibited a rapid burst release followed by a sustained release. 17-AAG-PBCA-NPs showed good stability in plasma. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay of the 17-AAG-PBCA-NPs and 17-AAG using HepG2, A375, HeLa, MCF-7 and SHSY5Y cell line showed that the 17-AAG-PBCA-NPs and 17-AAG inhibited the growth of HepG2, A375, HeLa, MCF-7 and SHSY5Y cells, showing a time-dependent manner, respectively. Furthermore, in vivo anti-tumor activity of 17-AAG-PBCA-NPs was evaluated in sarcoma bearing mice following intraperitoneal injection. Compared with 17-AAG, 17-AAG-PBCA-NPs achieved superior sustained-release effect, and extended the dosing interval further, increased tolerated dose, reduced the side effect of the drug, and improved the compliance to medication, safety and medication. Conclusion 17-AAG-PBCA-NPs were prepared by using interfacial polymerization method. Further optimization of the preparation method is both beneficial to the characteristic and sustained release effect of 17-AAG-PBCA-NPs.