Kuziv I. Covalent labeling of biomolecules with the reagents based on 3-hetarylcoumarins

Carboxyalkyl-modified 3-hetarylcoumarins and their conjugates with biomolecules. Synthesis of the functionalized derivatives of 3-hetarylcoumarins as reagents for the covalent labeling of biomolecules, preparation of the fluorescent conjugates of biomolecules of various classes and study of spectral properties of the synthesized dyes and conjugates. Organic synthesis, adsorption, ion-exchange and reverse-phase column chromatography, gel filtration, thin-layer chromatography, 1Н and 13С NMR spectrometry, chromato-mass-spectrometry, absorption and fluorescence spectroscopy, confocal microscopy. The synthetic methods were developed to obtain a series of new reagents based on 3-hetarylcoumarins with intense emission in the blue spectral region for biomolecular labeling, including the water-soluble dye derivatives. Convenient methods of labeling of biomolecules of various classes by these reagents were developed. In particular, the methods of the direct labeling of nucleosides without their preliminary functionalization, the method of preparation of the conjugates of hydrophobic peptide L-trileucine, the methods of synthesis of fluorescently labeled oligonucleotides, including the (2'-5')-triadenylate, were proposed. The possibility of live cells visualization using the conjugates of D-glucosamine was demonstrated. The spectral-fluorescent properties of the obtained compounds were studied in methanol and phosphate buffer (pH 1.9-10.5). Fields of application – bioorganic chemistry, biomedical sciences.