Xanthine Nucleosides with Pyrazolo[3,4-d]pyrimidine Skeleton: Functionalization with Halogen Atoms, Clickable Side Chains, Pyrene, and iEDDA Cycloadducts, and Impact of Ionic Forms on Photophysical Properties.

Xanthine nucleosides play a significant role in the expansion of the four-letter genetic code. Herein, 7-functionalized 8-aza-7-deazaxanthine ribo- and 2'-deoxyribonucleosides are described. 2-Amino-6-alkoxy nucleosides were converted to halogenated 8-aza-7-deazaxanthine nucleosides by deamination followed by hydroxy/alkoxy substitution. 8-Aza-7-deaza-7-iodo-2'-deoxyxanthosine served as an intermediate in Suzuki-Miyaura, Sonogashira, and Stille reactions. Alkynyl and vinyl side chains as well as fluorescent tags were introduced. Pyrene conjugates were derived by copper(I)-catalyzed cycloaddition. Inverse-electron-demand Diels-Alder reaction of 8-aza-7-deaza-7-vinyl-2'-deoxyxanthosine with 3,6-dipyridyl-tetrazine proceeded with a second-order rate constant of 0.042 L M-1 s-1. X-ray analysis of 8-aza-7-deaza-7-vinyl-2'-deoxyxanthosine displayed two conformers with a syn conformation. Crystal packing is stabilized by xanthine base pairs. UV spectroscopy confirmed the sensitivity of 7-functionalized 8-aza-7-deazaxanthine nucleosides to pH changes. Halogen and alkynyl substituents decrease pK values, and vinyl, pyrene, or benzofuran leads to an increase. Fluorescence measurements of 8-aza-7-deaza-7-benzofuran-2'-deoxyxanthosine disclosed solvatochromism and enhanced fluorescence when the pH or the viscosity of the solvent is increased. Nucleoside pyrene conjugates connected by a linear linker displayed monomer emission, and two pyrene residues connected by a dendritic linker exhibited excimer emission. According to their fluorescence properties and sensitivity to pH changes, the functionalized 8-aza-7-deazaxanthine nucleosides expand the class of nucleosides applicable to fluorescence detection for diagnostic and therapeutic purposes.