Comparison of recombinant and synthetically formed monoclonal antibody-beta-lactamase conjugates for anticancer prodrug activation.
Conjugates of the L49 monoclonal antibody (binds to the p97 antigen on melanomas and carcinomas) were formed by attaching Enterobacter cloacae beta-lactamase (bL) to the L49-Fab' fragment using a heterobifunctional cross-linking reagent or by linking the enzyme to L49-sFv using DNA recombinant technology. The conjugates thus formed, L49-Fab'-bL and L49-sFv-bL, were designed to activate cephalosporin containing anticancer prodrugs at the surfaces of antigen positive tumor cells. Results from in vitro experiments using two lung carcinoma cell lines demonstrated that the conjugates were equally active in effecting the release of phenylenediamine mustard from the cephalosporin nitrogen mustard prodrug CCM. While treatment with either of the conjugates combined with the maximum tolerated doses of CCM led to cures of established SN12P renal cell carcinoma tumors in nude mice, only the L49-sFv-bL conjugate maintained its ability to do so at 1/4 the maximum tolerated dose of CCM. L49-sFv-bL was also superior to L49-Fab'-bL in the 1934J renal cell carcinoma tumor model and was shown to be quite active in two in vivo models of human lung carcinoma. These results demonstrate that the recombinant fusion protein leads to more pronounced therapeutic windows than the chemical conjugate and is active in an array of human tumor models.
Kerr
D E
DE
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121, USA.
Vrudhula
V M
VM
Svensson
H P
HP
Siemers
N O
NO
Senter
P D
PD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000032:analysis / Q000378:metabolism; D000970:Antineoplastic Agents / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000188:drug therapy / Q000276:immunology; D002511:Cephalosporins / Q000378:metabolism / Q000627:therapeutic use; D006801:Humans; D007680:Kidney Neoplasms / Q000188:drug therapy / Q000276:immunology; D008175:Lung Neoplasms / Q000188:drug therapy / Q000276:immunology; D008545:Melanoma / Q000276:immunology; D058950:Melanoma-Specific Antigens; D051379:Mice; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000032:analysis / Q000378:metabolism; D009368:Neoplasm Transplantation; D009588:Nitrogen Mustard Compounds / Q000378:metabolism / Q000627:therapeutic use; D011355:Prodrugs / Q000378:metabolism*; D011994:Recombinant Proteins; D014407:Tumor Cells, Cultured; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D002511:Cephalosporins; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D009588:Nitrogen Mustard Compounds; D011355:Prodrugs; D011994:Recombinant Proteins; C038005:phenylenediamine mustard; D001618:beta-Lactamases