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A systematic approach, which combines previously reported reactions with appropriate modifications, was established for preparing a variety of selectively deuterated nucleosides from glucose. We have developed a systematic method of synthesizing specifically deuterated nucleosides from glucose. Selectively 13C/2H - doubly labeled nucleosides, such as [1′,2′,3′,4′,5′-13C5;3′,5′-2H2]- and [1′,2′,3′,4′,5′-13C5;4′,5′-2H2]-adenosines, have been synthesized starting from [ul-13C6]-d-glucose. These labeled nucleosides are very useful for precise conformational analyses of nucleic acids in solution by heteronuclear multidimensional NMR spectroscopy.

(5′ S)-[5′- 2H 1;1′,2′,3′,4′,5′- 13C 5]-Thymidine has been synthesized by a stereoselective deuteride transfer reaction from (−)- or (+)-[2- 2H 1]-isobornyloxymagnesium bromide to a 5-oxoribose derivative, which can be readily prepared from [ 13C 6]- d-glucose. The overall yield from d-glucose to thymidine was 27%. The various nucleosides with a stereoselective 2H-label together with 13C at the C5′ position, which have become available by the present method, will be quite useful for stereospecific assignment of the diastereotopic C5′ methylene signals, and also for conformational analyses of the O5′C5′ bonds in nucleic acid oligomers.

Thymidine with the stereoselectively 2H/13C-Labeled sugar moiety, (2′<it>R</it>)(5′<it>S</it>)-[1′,2′,3′,4′,5′-l3C<inf>5</inf>;2′,5′-2H<inf>2</inf>]-thymidine, was synthesized from uniformly 13C-abeled glucose, <it>via</it> the selectively deuterated ribose derivative prepared by the stereo-controlled deuteride transfer reactions. The labeled sugar moiety of the thymidine was then transferred to 2′-deoxyadenosine, 2′-deoxyguanosine, and 2′-deoxyuridine, by the enzymatic transglycosylation reactions by purine and pyrimidine nucleoside phosphorylases, in good yields. Labeled 2′-deoxyuridine was chemically converted to 2′-deoxycytidine. Consequently, all of the 2′-deoxynucleosides prepared by this method has the identically labeled sugar moiety. By using DNA oligomers containing the identically labeled sugar residue for NMR studies, any possible complexity in NMR data analyses expected to be observed for DNA oligomers containing variously labeled nucleosides can be minimized.