Tumor-selective drug activation: a GDEPT approach utilizing cytochrome P450 1A1 and AQ4N.
Drug metabolizing transgene products, which activate bioreductive cytotoxins, can be used to target treatment-resistant hypoxic tumors. The prodrug AQ4N is bioreduced in hypoxic cells by cytochrome P450s (CYPs) to the cytotoxin AQ4. Previously we have shown that intra-tumoral injection of CYP3A4 and CYP2B6 transgenes with AQ4N and radiation inhibits tumor growth. Here we examine the ability of other CYPs, in particular CYP1A1, to metabolize AQ4N, and to enhance radiosensitization. Metabolism of AQ4N was assessed using microsomes prepared from baculovirus-infected cells transfected with various CYP isoforms. AQ4N metabolism was most efficient with CYP1A1 (66.7 nmol/min/pmol) and 2B6 (34.4 nmol/min/pmol). Transient transfection of human CYP1A1+/-CYP reductase (CYPRED) was investigated in hypoxic RIF-1 mouse cells in vitro using the alkaline comet assay. There was a significant increase in DNA damage following transient transfection of CYP1A1 compared to non-transfected cells; inclusion of CYPRED provided no additional effect. In vivo, a single intra-tumoral injection of a CYP1A1 construct in combination with AQ4N (100 mg/kg i.p.) and 20 Gy X-rays caused a 16-day delay in tumor regrowth compared to tumors receiving AQ4N plus radiation and empty vector (P=0.0344). The results show the efficacy of a CYP1A1-mediated GDEPT strategy for bioreduction of AQ4N.
Yakkundi
A
A
Biomedical Sciences Research Institute, University of Ulster, Coleraine, UK.
McErlane
V
V
Murray
M
M
McCarthy
H O
HO
Ward
C
C
Hughes
C M
CM
Patterson
L H
LH
Hirst
D G
DG
McKeown
S R
SR
Robson
T
T
D000818:Animals; D000880:Anthraquinones / Q000378:metabolism / Q000494:pharmacology*; D015153:Blotting, Western; D015687:Cell Hypoxia / Q000187:drug effects; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D004249:DNA Damage / Q000187:drug effects; D004273:DNA, Neoplasm / Q000187:drug effects*; D015316:Genetic Therapy / Q000379:methods*; D051379:Mice; D015394:Molecular Structure; D011355:Prodrugs / Q000378:metabolism / Q000494:pharmacology*; D011838:Radiation-Sensitizing Agents / Q000378:metabolism / Q000494:pharmacology; D011878:Radiotherapy; D014407:Tumor Cells, Cultured
D000880:Anthraquinones; D004273:DNA, Neoplasm; D011355:Prodrugs; D011838:Radiation-Sensitizing Agents; C094428:AQ4N; D019363:Cytochrome P-450 CYP1A1