A Qualitative and Quantitative Assay to Study DNA/Drug Interaction Based on Sequence Selective Inhibition of Restriction Endonucleases

作者：Hassan Syed A^*; Chauhan Lata; Barthwal Ritu; Dixit Aparna

来源：Tropical Journal of Pharmaceutical Research, 2012, 11(5): 721-727.

DOI：10.4314/tjpr.v11i5.4

• Restriction endonucleases
• Restriction sites
• Anticancer drugs
• Restriction inhibition assay (RIA)
• Binding specificity

Purpose: To explore the use of restriction inhibition assay (RIA) to study the binding specificity of some anticancer drugs. %26lt;br%26gt;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. %26lt;br%26gt;Results: Mitoxantrone, adriamycin and daunomycin showed specificity for HindIII restriction site (5%26apos;-AAGCTT-3%26apos;) at 220, 100 and 100 mu M concentration, respectively. Conversely, distamycin-A showed an affinity for EcoRI (5%26apos;-AAATGC-3%26apos;) restriction sites at a concentration of 10 mu M. No binding was observed for berberine and palmatine at a maximum concentration of 2 mM at HindIII, EcoRI and EcoRV restriction sites, respectively. %26lt;br%26gt;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.

• 出版日期2012-10