Decreased cytotoxicity and increased antimitotic activity of a proline analogue of chlorambucil as a prodrug susceptible to the action of fibroblast's prolidase.
We synthesized an proline analogue of chlorambucil (CH-pro) as a prodrug susceptible to the action of ubiquitously distributed, cytosolic imidopeptidase--prolidase [E.C.3.4.13.9]. A conjugation of chlorambucil (CH) with proline through an imido-bond resulted in the formation of a good substrate for prolidase. We have compared several aspects of biological actions of CH and its prodrug in cultured normal human skin fibroblasts. The prodrug was found to be more effectively transported into the cells than the free drug. Moreover, in opposition to CH, CH-pro had no inhibitory effect on fibroblast's prolidase activity against the endogenous substrate, glycyl-L-proline. Lower cytotoxicity and a higher antimitotic activity of the prodrug, compared to the free drug, was observed. CH and CH-pro at concentrations of 25 microM led to a 30% and 10%, decrease in cell viability in confluent human skin fibroblasts. IC50 values of CH and CH-pro for DNA synthesis was found to be 30 microM and 7 microM, suggesting higher antimitotic potency of the pro-drug compared to the free drug. CH-pro also evoked lower ability to inhibit collagen biosynthesis in cultured fibroblasts than the free drug. IC50 values of CH and CH-pro for collagen biosynthesis were found at about 15 microM and 30 microM, respectively. Targeting of prolidase as a prodrug-converting enzyme may serve as a novel strategy in pharmacotherapy of various diseases, leading to the increase in therapeutic efficacy and reduction in untoward side effects of antineoplastic agents.
Bielawska
A
A
Department of Medicinal Chemistry, Medical Academy of Białystok, Białystok, Poland.
Chrzanowski
K
K
Bielawski
K
K
Pałka
J
J
D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D018906:Antineoplastic Agents, Alkylating / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002699:Chlorambucil / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology*; D004273:DNA, Neoplasm / Q000187:drug effects; D004150:Dipeptidases / Q000096:biosynthesis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D006801:Humans; D008934:Mitogens / Q000494:pharmacology; D011355:Prodrugs / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*
D000970:Antineoplastic Agents; D018906:Antineoplastic Agents, Alkylating; D004273:DNA, Neoplasm; D008934:Mitogens; D011355:Prodrugs; D002699:Chlorambucil; D004150:Dipeptidases; C022449:proline dipeptidase