载多柔比星人血清白蛋白毫微粒和两种免疫毫微粒的体内外靶向性研究

Doxorubicin-loaded nanoparticles and two kinds of immunonanoparticles:in vitro and in vivo targeting to tumor

目的构建载多柔比星的普通毫微粒、全抗体免疫毫微粒和抗体F(ab')2片段免疫毫微粒,比较它们对肿瘤的体外和体内靶向性,寻找对肿瘤组织具有特异性靶向作用的制剂。方法采用异型双功能交联剂琥珀酰亚胺基-3-(2-吡啶二硫)丙酸酯(SPDP),将兔抗大鼠乳腺癌细胞系Walker-256细胞的多克隆抗体(Wab)和其F(ab')2片段即WF(ab')2与载多柔比星(Doxorubicin,DRB)的人血清白蛋白毫微粒(DRB-HSA-NP)交联,制备全抗体免疫毫微粒Wab-DRB-HSA-NP和F(ab')2片段免疫毫微粒WF(ab')2-DRB-HSA-NP,比较它们的体外肿瘤细胞靶向性和体内组织器官靶向性。结果与DRB-HSA-NP相比,两种免疫毫微粒均有较好的体外肿瘤特异靶向性,瘤体内给药后在瘤体内的滞留量也显著增高,特别是WF(ab')2-DRB-HSA-NP,由于所偶联的抗体切除了FC段,降低了抗体与巨噬细胞的结合。结论免疫毫微粒,特别是WF(ab')2-DRB-HSA-NP具有较好的肿瘤靶向性,是一种很有价值的靶向制剂。

Objective To construct two kinds of doxorubicin- loaded immunonanoparticles: immunonanoparticles with complete antibody(Wab- DRB- HSA- NP), immunonanoparticles with F(ab′ )2 fragments 〔 WF(ab′ )2- DRB- HSA- NP〕 and investigate their targeting to tumor.Methods Hetero bi functional cross linker SPDP was used to couple anti Walker 256 cells polyclonal antibodies (Wab) or its F(ab′ )2 fragment 〔 WF(ab′ )2〕 with doxorubicin- loaded human serum albumin nanoparticles (DRB HSA NP).The targeting of nanoparticles (DRB HSA NP) and two kinds of immunonanoparticles to carcinoma cells was studied in vitro and in vivo.Results Compared with DRB- HSA- NP,both kinds of immunonanoparticles could efficiently bind to Walker- 256 cells in vitro. Although only a few of immunonanoparticles was detected in tumor tissue after administration via tail vein,the immunonanoparticals were mainly accumulated in tumor after intra- tumor administration.WF(ab′ )2 DRB HSA NP has shown better targeting activities than Wab- DRB- HSA- NP,as the F(ab′ )2 fragment has no Fc portion which can bind to Fc receptor on macrophages.Conclusions Immunonanoparticles can target to tumor,and antibody F(ab′ )2 fragment targeted immunonanoparticals are superior to antibody targeted immunonanoparticles.Intra tumor administration of immunonanoparticals yield a valuable therapeutic modality.