RGD肽修饰紫杉醇聚合物纳米粒的制备及其药效学研究

Preparation of paclitaxel-loaded polymeric nanoparticles and evaluation on their anti-tumor activities

制备靶向肽c(RGDyK)修饰的紫杉醇聚合物纳米粒,并对其体内外药效学性质进行评价。采用透析法制备靶向肽修饰的包载紫杉醇(PTX)的低相对分子质量肝素-全反式维甲酸聚合物(PTX-LHRyK)纳米粒,测定其粒度分布、Zeta电位、载药量和包封率等理化性质,通过体外细胞毒实验和体内药效学实验评价PTX-LHRyK纳米粒的抗肿瘤效果。制得的PTX-LHRyK纳米粒的粒径为(131.7±2.3)nm,Zeta电位为(-27.1±2.3)mV,载药量和包封率分别为(32.03±0.11)%和(84.84±2.63)%。随着孵育时间的延长,PTX-LHRyK纳米粒对B16F10细胞的毒性增加,孵育72 h后对B16F10细胞的IC50为(41.6±7.2)ng/mL,LHRyK载体对B16F10细胞的存活率无显著影响。体内药效学研究显示,PTX-LHRyK纳米粒的抑瘤率达到75.28%,是混合药物溶液组的1.46倍,纳米粒制剂组的小鼠体重和相对脾重均无显著性变化。因此,PTX-LHRyK纳米粒粒径小,载药量高,可明显提高紫杉醇的抗肿瘤治疗效果,且降低药物的不良反应。

The aim of the research was to prepare paclitaxel-loaded low molecular weight heparin-all-trans-retin- oid acid conjugate grafted with c（RGDyK） （PTX-LHRyK） nanoparticles, and to study the in vivo and in vitro an- ti-tumor effect of the PTX-LHRyK nanoparticles. The PTX-LHRyK nanoparticles were prepared by dialysis and characterized in terms of size, Zeta potential, and drug entrapment efficiency. The effect of PTX-LHRyK nanopar- ticles on PTX-induced cytotoxicity was investigated in B16F10 cell lines by MTI＂ assay. The in vivo anti-tumor effect （ in terms of tumor growth） and tumor inhibition rate were also evaluated. The PTX-LHRyK nanoparticles were generally spherical with a mean diameter of （ 131.7 ±2. 3） nm, a negative surface charge, and encapsulation efficiency of （84. 84 ±2. 63）%. The tumor inhibition rate of the PTX-LHRyK nanoparticles was up to 75.28%, which is 1.46-fold higher than the combined injection. It was shown in the in vitro anti-tumor activity study that the PTX-LHRyK nanoparticles displayed slightly higher in vitro cytotoxicity as compared to the combined injec- tion as the increase of the incubate time, but LHRyK conjugate at the studied concentration range did not show significant influence on the cell viability of the B16F10 cells. The in vivo anti-tumor effect of the PTX was signifi- cantly improved using the LHRyK conjugate as a carrier, with more powerful anti-tumor activity than PTX-ATRA- INJ as well as lower side-effects.