【作者】 於得红； 陆琴； 谢静； 方超； 陈红专；

【Author】 De-Hong Yu, Qin Lu, Jing Xie, Chao Fang*, Hong-Zhuan Chen* (Department of Pharmacology, Shanghai Jiao Tong University School of Medicine, 280 South Chongqing Road, Shanghai 200025, PR China)

【机构】 上海交通大学医学院药理学教研室；

【摘要】 目的构建一种载紫杉醇的靶向肿瘤血管内皮细胞的新型药物传释系统,探讨其体外、体内抗血管生成活性。方法采用开环聚合法合成aldehyde-PEG-PLA和MPEG-PLA聚合物。以两种聚合物一定比例的混合物为载体材料,采用乳化-溶剂挥发法制备装载紫杉醇的隐形纳米粒(PTX-NP)。K237多肽通过其末端氨基(NH2)与纳米粒表面PEG末端醛基共价连接得到装载紫杉醇的K237多肽修饰的隐形纳米粒(K237-PTX-NP)。采用人脐静脉内皮细胞体外模型和荷瘤小鼠体内模型研究隐形纳米粒的抗血管生成活性。结果K237-PTX-NP粒径为150nm(DLS检测),Zeta电位为-20mV。X-射线光电子能谱(XPS)分析确证纳米粒表面连接有K237多肽。高效液相色谱(HPLC)分析测定纳米粒载药量为2.8%,包封率为33.5%。细胞学实验表明,在PTX浓度为5×10-5-50nM的试验范围内,K237-PTX-NP抑制HUVEC增殖的活性显著大于PTX-NP,并在PTX浓度大于0.05nM时显著高于市售紫杉醇注射液(Taxol);K237-PTX-NP在PTX浓度小于0.005nM时抑制HUVEC迁移的活性显著大于PTX-NP,并在全部浓度范围内(5nM除外)显著大于Taxol;K237-PTX-NP在PTX浓度大于0.05nM时抑制HUVEC小管形成的活性显著大于PTX-NP,并在PTX浓度为0.5和5nM时显著大于Taxol。与Taxol相比,K237-PTX-NP具有"隐形"特性,所装载的药物可在循环系统中长期滞留,通过K237-KDR介导的内吞作用实现肿瘤血管内皮细胞靶向抗血管生成作用。体内荷瘤动物实验表明,K237-PTX-NP可特异性靶向肿瘤新生血管,诱导肿瘤血管内皮细胞凋亡和肿瘤组织坏死。结论K237-PTX-NP可特异性靶向肿瘤新生血管,实现抗肿瘤血管生成治疗肿瘤的目的。更多还原

【Abstract】 OBJECTIVE To develope a novel drug delivery system, stealth nanoparticles decorated with K237 peptides (K237-NP), which could target to the tumor neovasculature. METHODS Aldehyde poly (ethylene glycol)-poly (lactide) (CHO-PEG-PLA) and methoxy poly (ethylene glycol)-poly (lactide) (CH3O-PEG-PLA) copolymers were synthesized by the ring opening polymerization. The PTX-NP were prepared with a composition-optimized blend of CHO-PEG-PLA and CH3O-PEG-PLA using the O/W emulsion-evaporation technique. K237-PTX-NP were obtained by conjugating the N-terminal amine of K237 molecules to the aldehyde of PEG on the surface of PTX-NP. The HUVEC and the tumor-loaded models were used to evaluate the in vitro and in vivo antiangiogenic activity of K237-PTX-NP. RESULTS The particle size of K237-PTX-NP was around 150 nm (DLS assay) and the Zeta potential was about-20 mv. X-ray photoelectron spectroscopy (XPS) analysis confirmed the existence of K237 molecules on the particle surface. The drug loading was 2.8% and the encapsulation efficiency was 33.5% determined using the high performance liquid chromatography (HPLC). With courmarin-6 as the fluorescent probe, the observation under the confocal laser scanning microscopy (CLSM) showed that K237 peptides decorated on the particle surface significantly facilitated the particle uptake by HUVEC. In the experiment range (PTX concentration of 5×10-5-50 nM), K237-PTX-NP caused dramatically higher cytotoxicity to HUVEC than PTX-NP at all tested concentrations and than marketed paclitaxel injection (Taxol) at drug concentration of more than 0.05 nM. K237-PTX-NP exhibited much higher activity of inhibiting HUVEC migration than PTX-NP at drug concentration of less than 5×10-3 nM and than Taxol at all tested concentrations except 5 nM. K237-PTX-NP displayed higher activity of inhibiting tubule formation than PTX-NP at drug concentration of above 0.05 nM and Taxol at paclitaxel concentration of 0.5 and 5 nM. With the ‘stealth’ property, K237-PTX-NP could extend the circulating time of encapsulated drug, acquire an enhanced uptake by HUVEC through the binding of K237 and KDR, and achieve the antiangiogenic therapy. The in vivo experiment showed that K237-PTX-NP could specifically target to the tumor neovasculature, induce the apoptosis of tumor endothelial cells and tumor necrosis. CONCLUSION K237-PTX-NP can specifically target to the tumor neovasculature and achieve the antiangiogenic cancer therapy.更多还原