Synthesis and characterization of a catalytic antibody-HPMA copolymer-Conjugate as a tool for tumor selective prodrug activation.
Selective chemotherapy remains a key issue for successful treatment in cancer therapy. The use of targeting approaches like the enhanced permeability and retention (EPR) effect of macromolecules, is consequently needed. Here, we report the preparation of a novel catalytic antibody-polymer conjugate for selective prodrug activation. HPMA copolymer was conjugated to catalytic antibody 38C2 through an amide bond formation between epsilon-amino group of lysine residue from the antibody molecule and a p-nitrophenyl ester of the polymer. The conjugate was purified over a size exclusion column using FPLC. In the isolated fraction, one or two molecules of polymer were conjugated to one molecule of antibody based on gel analysis. The resulting conjugate retained most of its catalytic activity (75-81%) in comparison to the free antibody. The activity was monitored with a fluorogenic substrate and a prodrug activation assay using HPLC. Furthermore, the conjugate was evaluated in vitro for its ability to activate an etoposide prodrug using two different cancer cell lines. Cells growth inhibition using the prodrug and the conjugate was almost identical to inhibition by the free antibody and the prodrug. For the first time, a catalytic antibody was conjugated to a passive targeting moiety while retaining its catalytic ability to activate a prodrug. The conjugate described in this work can be used for selective activation of prodrug in the PDEPT (polymer directed enzyme prodrug therapy) approach by replacing the enzyme component with catalytic antibody 38C2.
Department of Cell Research and Immunology, Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
Ronit
R
Satchi-Fainaro
Wolfgang
W
Wrasidlo
Holger N
HN
Lode
Doron
D
Shabat
D017151:Antibodies, Catalytic / Q000737:chemistry / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D005047:Etoposide / Q000493:pharmacokinetics; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D008689:Methacrylates / Q000737:chemistry / Q000494:pharmacology; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology; D011355:Prodrugs / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured
D017151:Antibodies, Catalytic; D018796:Immunoconjugates; D008689:Methacrylates; D011355:Prodrugs; D005047:Etoposide; C032802:hydroxypropyl methacrylate