Specific chemothrapy for perineural invasion of biliary tract cancer with conjugate of anticancer drug and anti-neural cell adhesion molecule antibody

制备了化疗药物丝裂霉素C （Mitomycin C，MMC）与单克隆抗体抗神经细胞粘附分子（NCAM）抗体的偶联物，并对其体外抗肿瘤活性和抗体活性进行了评价。根据Suzuki等人的方法，用溴化氰（BrCN）法制备MMC与抗ncam抗体偶联物。假设MMC的吸光度为λ max=363nm, E1%1cm=623， IgG的吸光度为λ max=280nm, E1%/1cm=13.5，用分光光度法测定取代度。取代度计算公式为：取代度=偶联物中所含药物的摩尔浓度/偶联物中所含抗体的摩尔浓度，平均为3.5。用MTT法测定了偶联物对神经母细胞瘤细胞株（IMR-32）的生长抑制作用，与相同浓度的MMC溶液相比，偶联物的药物活性保持在66 ~ 95%。用avidin -生物素化酶复合物对NCAM阳性胆管癌进行免疫组化染色检测，偶联物的抗体活性与单独抗体水平相同。综上所述，MMC可以通过BrCN方法与抗ncam单克隆抗体连接。由于该偶联物具有药物活性和抗体活性，可为特异性靶向肿瘤化疗提供有用的工具。

Conjugate of chemotherapeutic drug (Mitomycin C,MMC) and monoclonal antibody, which was anti-Neural cell adhesion molecule (NCAM) antibody was prepared and evaluated anticancer activity and antibody activity of the conjugate in vitro. MMC and anti-NCAM antibody conjugate was prepared according to Suzuki et al with cyanogen bromide (BrCN) method.The degree of substitution was determined spectrophotometrically by assuming that MMC has absorbance with lambdamax=363nm, E1%1cm=623 and IgG has absorbance with lambda max=280nm, E1%/1cm=13.5. The degree of substitution which was calculated as follows : degree of substitution=mol concentration of drug contained in conjugate/mol concentration of antibody contained in conjugate, was 3.5 (average).A drug activity of conjugate, which was tested by MTT method of the growth inhibition with neuroblastoma cell line (IMR-32) was maintained 66 to 95% compared with same concentration of MMC solution.Antibody activity of conjugate, which was examined by immunohistochemical staining with Avidin-Biotinylated enzyme Complex on NCAM positive bile duct cancer, showed the same level as that of antibody alone.In conclusion, MMC could be linked to anti-NCAM monoclonal antibody by BrCN method. Since this conjugate has both activity of drug and antibody, it can provide an useful tool for specific targeting cancer chemotherapy.