title: Enhancement of antibody-directed enzyme prodrug therapy in colorectal xenografts by an antivascular agent.
abstract: The irregular nature of solid tumor vasculature produces a heterogeneous distribution of antibody-targeted therapies within the tumor mass, which frequently results in reduced therapeutic efficacy. We have, therefore, combined two complementary therapies: Antibody-directed Enzyme Prodrug Therapy (ADEPT), which targets tumor cells, and an agent that selectively destroys tumor vasculature. A single i.p. dose (27.5 mg/kg) of the drug 5,6-dimethylxanthenone-4-acetic acid (DMXAA), given to nude mice bearing the LS174T colorectal xenograft, destroyed all but a peripheral rim of tumor cells, without enhancing survival. The ADEPT system, in which a pretargeted enzyme activates a prodrug, consisted of the F(ab')2 fragment of anti-carcinoembryonic antigen antibody A5B7 conjugated to the bacterial enzyme carboxypeptidase G2 and the prodrug 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-L-glutamic acid, which was given i.p. in three doses of 500 mg/kg at 72, 84, and 96 h post-conjugate administration (25 units of carboxypeptidase G2). The antibody-enzyme conjugate could be selectively retained at approximately twice the control levels by administration of the antivascular agent at the time of optimal conjugate localization within the tumor (20 h post-conjugate administration), as demonstrated by gamma counting, phosphor plate image analysis, and active enzyme measurement. This resulted in significantly enhanced tumor growth inhibition in groups of six mice, compared to conventional ADEPT therapy, with no concomitant increase in systemic toxicity. In a separate experiment, aimed at trapping the prodrug within the tumor, a 16-fold increase over control values was produced (means, 44.8 versus 2.8 microg/g tumor) when DMXAA was given 4 h prior to 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-L-glutamic acid. The therapeutic window was small, with no significant enhancement of prodrug retention when DMXAA was given at either earlier or later time points. This correlated with the time of vascular shut-down induced by the antivascular agent. We are currently investigating whether it is more advantageous to trap increased levels of conjugate or prodrug within the tumor for maximal enhancement of conventional ADEPT. These studies demonstrate that combined use of antibody-directed and antivascular therapies can significantly benefit the therapeutic outcome of either strategy alone.
authors.affiliation: Department of Oncology, Royal Free and University College Medical School, University College London, United Kingdom.
authors.forename: R B
authors.initials: RB
authors.lastname: Pedley
authors.forename: S K
authors.initials: SK
authors.lastname: Sharma
authors.forename: G M
authors.initials: GM
authors.lastname: Boxer
authors.forename: R
authors.initials: R
authors.lastname: Boden
authors.forename: S M
authors.initials: SM
authors.lastname: Stribbling
authors.forename: L
authors.initials: L
authors.lastname: Davies
authors.forename: C J
authors.initials: CJ
authors.lastname: Springer
authors.forename: R H
authors.initials: RH
authors.lastname: Begent
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D002272:Carcinoembryonic Antigen / Q000276:immunology; D015179:Colorectal Neoplasms / Q000098:blood supply / Q000188:drug therapy* / Q000276:immunology; D005260:Female; D005971:Glutamates / Q000008:administration & dosage*; D006801:Humans; D018796:Immunoconjugates / Q000008:administration & dosage*; D007140:Immunoglobulin Fab Fragments / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D009588:Nitrogen Mustard Compounds / Q000008:administration & dosage*; D011355:Prodrugs / Q000008:administration & dosage*; D014183:Transplantation, Heterologous; D014966:Xanthenes / Q000008:administration & dosage*; D044004:Xanthones; D011623:gamma-Glutamyl Hydrolase / Q000008:administration & dosage
chemical_list: D000970:Antineoplastic Agents; D002272:Carcinoembryonic Antigen; D005971:Glutamates; D018796:Immunoconjugates; D007140:Immunoglobulin Fab Fragments; D009588:Nitrogen Mustard Compounds; D011355:Prodrugs; D014966:Xanthenes; D044004:Xanthones; C066668:vadimezan; C092574:4-((2-chloroethyl)(2-mesyloxyethyl)amino)benzoylglutamic acid; D011623:gamma-Glutamyl Hydrolase