Maltsev G. Application of the luminescent method of analysis to study the interaction of some drugs with DNA and proteins

The thesis is devoted to the study of the interaction of some medicinal substances (MS) with DNA and proteins using the luminescence method of analysis. A new luminescent probe-terbium (III) complex with the 2-oxo-4-hydroxoquinoline-3-carboxylic acid derivative was proposed for the first time to study of the intercalation in the DNA of a number of aminoalkoxy fluorenones (AF). By the method of competition with a new lanthanide probe showed that all synthesized compounds are DNA intercalators, logarithms of the constants of association of synthesized compounds with DNA have been determined. The interaction of MS with HSA has been evaluated by fixing the changes in the intrinsic fluorescence intensity of the protein (quenching) with the addition of MS. Conformational changes in HSA caused by binding to MS by measuring synchronous fluorescence spectra have been studied. It is shown that changes in protein conformation near the tryptophan residue are observed practically for all MS and practically no change in the environment of the tyrosine residue. An approach has been developed that is to quench the intrinsic fluorescence of the ligand upon addition of the protein and the calculating apparatus for determining the binding parameters. In the case of MS (indomethacin, daclatasvir dihydrochloride), for which their emission spectrum is superimposed on the intrinsic fluorescence of the protein, the possibility of determining the binding constants of HSA with drug molecules by quenching their intrinsic fluorescence have been demonstrated. The highly sensitive methods of luminescent DNA determination using a luminescent probe Tb (III) -L2 (limit of detection 30 ng/ ml) and determination of residual quantities of propoxazepam on the surfaces of pharmaceutical equipment (limit of detection 0.5 μg/ml) have been developed and validated .