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

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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.