人肝癌特异性阿霉素人血清白蛋白免疫微球的鉴定及药剂学性质研究

Studies on preparation and pharmaceutical features of human hepatoma-s pecific immunomicrospheres

目的 制备抗体偶联的阿霉素人血清白蛋白微球 ;评价该免疫微球的药剂学性质。方法 采用SPDP法将人肝癌单抗HAb1 8与阿霉素人血清白蛋白微球 (ADR HSA MS)共价偶联构建人肝癌特异性免疫微球 (HAb1 8 ADR HSA MS) ;采用玻片凝集法 ,免疫荧光染色法评价抗体与微球的交联 ;采用胃酶消化法 ,动态透析法分别测定免疫微球的载药量 ,体外释药性质 ;采用MTT比色分析法测定免疫微球经胃酶消解所释放ADR的细胞毒作用。结果 HAb1 8 ADR HSA MS的玻片凝集反应 ,免疫荧光染色均为阳性 ;ADR的平均回收率为 (98.95± 0 .96) % ;HAb1 8 ADR HSA MS表面光滑 ,圆球状 ,平均粒径 1 .2 μm ,有效载药量 1 .44 % ,最大累积释药分数为 41 % ;HAb1 8 ADR HSA MS的胃酶消解液在 50 0nm波长处有最大吸收 ,对人肝癌细胞株SMMC 772 1的杀伤作用曲线相似于ADR HSA MS及游离ADR。结论 采用SPDP法能将人肝癌单抗HAb1 8结合到阿霉素人血清白蛋白微球表面 ;该交联对免疫微球的有效载药量 ,最大累积释药量有一定程度影响 。

OBJECTIVE To prepare the adrimycin loaded albumi n microspheres with antibody binding by SPDP conjugation technique and to evaluate the pharmaceutical properties of the immunomicrospheres against human hepatoma( HAb18 ADR HSA MS).METHODS Human hepatoma specific monocl onal antibody HAb18 was coupled to adrimycin loaded albumin microspheres(ADR H SA MS).Slide agglutination test and immunofluorescent assay were used for evalu ation of the antibody cross linking with ADR HSA MS.Pepsin digestion method w as used for determining drug loading of immunomicrospheres and membrane diffusi on technique for detection of in vitro release of ADR from the immunomicrosp heres.MTT colorimetric assay was used to determine cytotoxicities of ADR release d by immunomicrospheres for tumor cell lines.RESULTS HAb18 ADR HSA MS,red sphere with average diameter of 1.2 μm,was positive in slide a gglutination test and immunofluorescent assay.Its effective drug loading and pea k of the accumulated drug releasing fraction were 1.44% and 41% respectively,lo wer than those of ordinary microspheres(ADR HSA MS)(1.69%,65%).The pepsin dig estion solution of the HAb18 ADR HSA MS had a absorption peak at 500 nm wavel ength and its killing curve for human hepatoma SMMC7721 was similar to those of ADR HSA MS and free ADR.CONCLUSION Human hepatoma specifi c monoclonal antibody could be conjugated to ADR loaded albumin microspheres by SPDP conjugation method.The conjugation has remarkable effects on drug loading and drug release of immunomicrospheres,but little influences on its drug efficac y.