Bleomycin A6 (A6), a single component of
bleomycin complex, is highly active against human
liver cancer cells in vitro and xenografts in nude mice. A6 was conjugated to
monoclonal antibody H111 directed against human
hepatoma BEL - 7402 cells, using
Dextran T40 as an intermediate. The conjugate consisted of a coupling molar ratio of 1:264 for H111 and A6, and retained 6.3% of A6 activity. As determined by clonogenic assay with
hepatoma BEL - 7402 cells exposed to the agents for 1 h, the IC90 values for H111 - A6 conjugate, free A6 and M3 - A6 conjugate (an irrelevant conjugate) were 0.17 mu mol/L, 17 mu mol/L and 7 mu mol/L respectively. The cytotoxicity of Hill - A6 conjugate to target cells was markedly blocked by unconjugated H111 but not by irrelevant
monoclonal antibody M3. The H111 - A6 conjugate exhibited 78% inhibition on the growth of
hepatoma BEL - 7402 xenografts in nude mice, whereas the equivalent doses of free A6, M3 - A6 conjugate and H111 plus A6 mixture showed approximately 30% inhibition. Histopathological examination showed no toxic changes in the liver, lung, kidney and bone marrow in the H111 - A6 conjugate--treated animals. These results suggest that the conjugate of
monoclonal antibody and
bleomycin A6 exhibits specific cytotoxicity to target
liver cancer cells and the conjugate is highly effective against
liver cancer xenografts in nude mice with more marked
tumor inhibition than free A6 at comparable dose levels.