Target-selective cytotoxicity of methotrexate conjugated with monoclonal anti-MM46 antibody.
In studies on antitumor antibody-cytotoxic drug conjugates as potential tumor-selective cytotoxic agents, methotrexate (MTX) was conjugated via its active ester derivative with a murine monoclonal antibody (aMM46) to a mouse mammary tumor antigen (MM antigen) on syngeneic, ascitic C3H/He mouse mammary tumor MM46 cells. The conjugate retained full antibody activity, as assayed by complement-dependent cytolysis. The target-selective cytotoxicity of aMM46-MTX was verified by the observations that this conjugate showed greater cytotoxicity than the corresponding normal mouse immunoglobulin (nIg) conjugate to MM46 cells, neither aMM46 nor nIg being cytotoxic, and that it showed less cytotoxicity to MM antigen negative mouse mammary tumor MM48 cells than to MM46 cells, its cytotoxicity to MM48 cells being similar to that of the nIg conjugate. From the results of assays of cell binding and uptake of 131I-labeled aMM46 and aMM46-3H-MTX, aMM46 and aMM46-MTX were internalized after their binding to MM46 cell surface antigen. Leupeptin, an inhibitor of the lysosomal endopeptidase cathepsin, decreased the cytotoxicity of aMM46-MTX, supporting the involvement of lysosomal degradation of the conjugate in its action.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage*; D002460:Cell Line; D000922:Immunotoxins / Q000008:administration & dosage*; D007976:Leupeptins / Q000494:pharmacology; D008325:Mammary Neoplasms, Experimental / Q000628:therapy*; D008727:Methotrexate / Q000008:administration & dosage*; D051379:Mice; D008809:Mice, Inbred C3H
D000911:Antibodies, Monoclonal; D000922:Immunotoxins; D007976:Leupeptins; C032854:leupeptin; D008727:Methotrexate