新型姜黄素纳米粒制备、表征及其体外抗肿瘤活性评价

Preparation,Characteristic and Anti - tumor Activity Evaluation of Novel Curcumin Nanoparticle in vitro

目的制备高载药量姜黄素纳米粒,并考察其体外稳定性和抗肿瘤活性。方法用油酸(OA)对姜黄素(Cur)进行化学修饰。采用改良的溶剂挥发法制备聚乙二醇聚乳酸乙酸酯(mPEG-PLGA)载Cur-OA2纳米粒(mPEG-PLGA-Cur-OA2,PPCO)。并以纳米粒载药量(drug loading,DL)、包封率(entrapment efficiency,EN)为指标,通过3因素3水平正交试验对工艺进行优化。采用正交确定工艺制备3批载药纳米粒,应用动态光散射粒度仪和透射电镜测定载药纳米粒的zeta电位、粒径与形态。采用体外37℃水浴降解特性来评价其稳定性。最后利用MTT法对纳米粒体外抗肿瘤活性进行初步评价。结果正交实验,包封率影响因素为:有机相与水相的量(B)>超声时间(C)>药物与材料比(A)。载药量影响因素为:有机相与水相的量(B)>药物与材料比(A)>超声时间(C)。利用正交设计筛选出的方法制备纳米粒,其载药量达(24.870±0.029)%,包封率为(81.250±0.101)%,zeta电位-23.9±1.6mV,平均粒径235.0±25.8nm,粒度分布均匀,呈单峰分布。载药纳米粒在37℃,前4h降解了20%,而其后的70h里,只降解了5%左右,相比姜黄素稳定性得到了极大提高。纳米粒体外抗肿瘤活性研究表明,所制备的纳米粒对HepG2细胞仍然具有较好的抑制作用,经48h处理后,其IC50为40.61μmol/L,但相比姜黄素15.76μmol/L有所下降,表现为减毒效应。结论 PPCO纳米粒呈均匀球形、载药量高,稳定性好,并有较好的体外抗肿瘤活性。

Objective To prepare high Drug - loading curcumiin nanoparticles, and evaluate its stability and anti - cancer activity in vitro. Methods Curcumin （Cur） was chemical modified by oleic acid （OA） to obtain conjugate Cur - OA2. Modified solvent evapo- ration method was used to prepare mPEG - PLGA nanoparticles containing Cur - OA2 （ mPEG - PLGA - Cur - OA2 , PPCO）. Orthogonal test was adopted to optimize the preparation method with drug loading （DL） and entrapment efficiency （EN） as index. Then three batches of nanoparticles were prepared according the optimized method, and the zeta potential, particle size and morphology of the drug - loaded nanoparticles were analyzed by dynamic light scattering particle size analyzer （DLS） and transmission electron microscopy （TEM）. Finally, the stability and anti - tumor activity of the PPCO in vitro were evaluated by hydrolysis method and MTT assay respectively. Results Orthogonal test indicated that the influencing factor of EN was ：the amount of the organic and aqueous （B） 〉 ultrasonic time （C） 〉 drug and material ratio （A） ; and the influencing factor of DL was：the amount of the organic and aqueous （B） 〉 drug and material ratio （A） 〉 ultrasonic time （ C）. The PPCO was prepared by optimized method. DLS and TEM revealed that the nanoparticle was rounded and sin- gle distributed, with drug loading （24. 870 ± 0. 029） % , entrapment efficiency （ 81. 250 ± 0. 101 ） % , zeta potential （ - 23.9 ± 1.6 ） mV and average particle size （235.0 ± 25.8） nm. PPCO was unstable in 4h, and 20% loaded drug was degradated, but it became very stable in the following 70h under 37℃ in water. Eventually, MTT confirmed PPCO still with great inhibition effect on HepG2 cell lines with IC50 =40.61μmol/L, lower than that of curcumin （15. 76μmol/L）. Conclusion PPCO nanoparticles had even spherical form, high drug - loading, good stability and great anti - tumor activity in vitro.