A novel and simple method of screening compounds for interaction with DNA: A validation study.
We report the development of a simple, cost-effective assay for detecting compounds that have the ability to interact with and modify DNA. Potential uses for the assay lie in the areas of early genotoxicity testing of drug candidates, anticancer and antibiotic drug discovery, environmental monitoring and testing in the food, beverage and cosmetics industries. At present the assay has been used to assess direct-acting compounds only and it is yet to be established whether the assay is compatible with bio-activation. The methodology is based on the oxidative reaction of potassium permanganate with pyrimidine bases, which have become perturbed and more reactive by the agent under test. Results are recorded by use of UV/vis spectroscopy. The adaptation to a multi-well plate format provides the capacity for high throughput utilizing small amounts of compounds. Over 100 compounds, comprising different classes of DNA-binding chemicals as well as non-binding controls, have been put through the assay and the results compared with existing genotoxicity testing data from other methods. The assay has shown to be predictive of the results of other genotoxicity testing methods. We have found that the method is overall predictive of 71% of Ames bacterial reverse-mutation test results (where data are given) encompassing both negative and positive results.
Garas
Adel
A
Genomic Disorders Research Centre, Alan Gilbert Building, 161 Barry Street, Carlton South, VIC 3053, Australia.
Webb
Elizabeth
E
Pillay
Vinochani
V
MacPhee
Donald
D
Denny
William
W
Zeller
Hilary
H
Cotton
Richard
R
D000477:Alkylating Agents / Q000633:toxicity; D004249:DNA Damage; D055808:Drug Discovery; D007364:Intercalating Agents / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D011196:Potassium Permanganate / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D000477:Alkylating Agents; D007364:Intercalating Agents; D009153:Mutagens; D011743:Pyrimidines; D011196:Potassium Permanganate; C030986:pyrimidine