title: Measurement of the critical DNA lesions produced by antibody-directed enzyme prodrug therapy (ADEPT) in vitro, in vivo and in clinical material.
abstract: An antibody-directed enzyme prodrug therapy (ADEPT) system against CEA-positive tumours is currently in phase I clinical trials. It consists of a prodrug, 4-[N,N-bis(2-iodoethyl) amino] phenoxycarbonyl L -glutamic acid (ZD2767P) and a conjugate of the F(ab')(2) anti-CEA antibody A5B7 and the bacterial enzyme carboxypeptidase G2 (CPG2). ZD2767P is converted by antibody-targeted CPG2 into an active bifunctional alkylating drug (ZD2767) at the tumour site. The IC(50) value of the prodrug against the human colorectal tumour LS174T cell line was 55 +/- 9 microM following a 1 h exposure. In contrast, co-incubation of ZD2767P with CPG2 resulted in 229-fold increase in activity. Using a modified comet assay, DNA interstrand cross links (ISC) were detected within 1 h of ZD2767P + CPG2 treatment and were repaired by 24 h. A clear dose-response was seen between the level of ISC, growth inhibition and ZD2767 concentration. Administration of a therapeutic dose of ZD2767P 72 h after the F(ab')(2) A5B7 conjugate to mice bearing LS147T xenografts resulted in extensive ISC in the tumour after 1 h; repair was seen at 24 h. Tumour biopsies and peripheral lymphocytes were studied in 5 patients on the ADEPT phase I clinical trial. In 4 patients no ISC were detected. These patients also demonstrated poor localization of conjugate and no tumour response was seen. However a significant level of ISC was detected in one tumour biopsy, which also showed evidence of conjugate localization and clinical response. These studies demonstrate the application of the comet assay in the measurement of ISC in vitro and in clinical material and confirm that activation of ZD2767P results in the formation of DNA crosslinks.
authors.affiliation: Department of Oncology, Royal Free and University College School of Medicine, University College London for the Phase I and II Clinical Trials Committee of the Cancer Research Campaign, UK.
authors.forename: S D
authors.initials: SD
authors.lastname: Webley
authors.forename: R J
authors.initials: RJ
authors.lastname: Francis
authors.forename: R B
authors.initials: RB
authors.lastname: Pedley
authors.forename: S K
authors.initials: SK
authors.lastname: Sharma
authors.forename: R H
authors.initials: RH
authors.lastname: Begent
authors.forename: J A
authors.initials: JA
authors.lastname: Hartley
authors.forename: D
authors.initials: D
authors.lastname: Hochhauser
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000627:therapeutic use*; D018906:Antineoplastic Agents, Alkylating / Q000276:immunology / Q000627:therapeutic use; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D004249:DNA Damage; D005260:Female; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007167:Immunotherapy; D008214:Lymphocytes; D051379:Mice; D009588:Nitrogen Mustard Compounds / Q000276:immunology / Q000627:therapeutic use; D011355:Prodrugs; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D011623:gamma-Glutamyl Hydrolase / Q000378:metabolism*
chemical_list: D000906:Antibodies; D018906:Antineoplastic Agents, Alkylating; D002272:Carcinoembryonic Antigen; D007140:Immunoglobulin Fab Fragments; D009588:Nitrogen Mustard Compounds; D011355:Prodrugs; C097602:ZD 2767; D011623:gamma-Glutamyl Hydrolase