Purpose: To explore the use of restriction inhibition assay (RIA) to study the binding specificity of some anticancer drugs. Methods: A 448 bp DNA fragment derived from pBCKS+ plasmid (harboring the polylinker region with multiple restriction endonuclease sites) was used as a template for sequence selective inhibition of the test drugs. The template was incubated with different concentrations of anticancer drugs (adriamycin, daunomycin, mitoxantrone, distamycin-A, berberine and palmatine) prior to digestion with restriction endonucleases - HindIII, EcoRI and EcoRV. Results: Mitoxantrone, adriamycin and daunomycin showed specificity for HindIII restriction site (5'- AAGCTT-3') at 220, 100 and 100 μM concentration, respectively. Conversely, distamycin-A showed an affinity for EcoRI (5'-AAATGC-3') restriction sites at a concentration of 10 μM. No binding was observed for berberine and palmatine at a maximum concentration of 2 mM at HindIII, EcoRI and EcoRV restriction sites, respectively. Conclusion: The inhibition of endonucleases by mitoxantrone, adriamycin, daunomycin, distamycin-A, provides direct evidence of the co-existence of concentration and sequence specificity for drug-DNA interaction as well as the need to explore the possible use of RIA for demonstrating the binding specificity of anticancer drugs. © Pharmacotherapy Group.
CITATION STYLE
Hassan, S. A., Chauhan, L., Barthwal, R., & Dixit, A. (2012). A qualitative and quantitative assay to study DNA/drug interaction based on sequence selective inhibition of restriction endonucleases. Tropical Journal of Pharmaceutical Research, 11(5), 721–727. https://doi.org/10.4314/tjpr.v11i5.4
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