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To develop an efficient method for the synthesis of a highly diasteroselective (2'R)-2'-deoxy[2'-2H]guanosine (1), studies of organic chemical conversion from 2'-bromo-2'-deoxy-N2-Isobutyryl-3',5'-O-TIPDS-guanosine (2) to 1 and a biological transdeoxyribofuranosylation of (2'R > 98% de)-2'-deoxy[2'-2H]uridine (4) were carried out. As the results, a highly diastereoselective synthesis of 1 was achieved by a biological transdeoxyribofuranosylation between 2,6-diaminopurine and 4 by the use of Enterobacter aerogenes AJ-11125, followed by treatment with adenosine deaminase. The results will be described in detail.

Nucleophilic substitution reaction of 6-chloro-2-fluoro-9-beta-D-ribofuranosyl-9H-purine derivative, prepared from guanosine, with potassium [15N]phthalimide at 40 degrees C for 9 h in DMF, followed by hydrolysis, afforded [2-(15)N]guanosine derivative efficiently. The corresponding 2'-deoxy derivative was also synthesized through a similar procedure.

Thymidine with the stereoselectively 2H/13C-Labeled sugar moiety, (2'R)(5'S)-[1',2',3',4',5'-(13)C5;2',5'-(2)H2]-thymidine, was synthesized from uniformly 13C-labeled glucose, via 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.

A "one-pot reaction" has been used for simultaneous protection of amino and O6-function of deoxyguanosine. N-(4-bromobutyl) phthalimide used for O6-protection generates an alkylamino function after mild base hydrolysis. Two sequences d(GGA) and d(GGATCC) have been synthesised with all protected guanosine units. Similarly, 2-(vinyl)pyridine has also been used for O6-protection following the same strategy.

We reinvestigated the reaction of dGuo in the presence of nitrous acid by reversed phase HPLC (RPLC) analysis. When 10 mM deoxyguanosine (dGuo) was treated with 100 mM NaNO2 in 3.0 M acetate buffer (pH 3.7) at 37 degrees C, unknown peak appeared in large amount (yield 19.7% at 2 h, 28.7% at 6 h). The yield of this newly found compound, denoted as 1, arose with increasing H+ concentration and was maximal at 50 degrees C. 1 has small depurination rate constant. If no enzymatic repair system for 1 exists, 1 would have strong mutagenic ability.

A method was developed to synthesize 2'-deoxy-8,2'-methylene-cycloadenosine (1) and -cycloguanosine (2) which were new carbon-bridged cyclopurine nucleosides fixed in a high-anti torsional angle region. 3',5'-Di-O-acetyl-8-methanesulfonyl-2'-O-p-toluene-sulfonyladenosine+ ++ (3) or 2-acetamido-9- (3,5-di-O-acetyl-2-O-p-toluenesulfonyl-beta-D-ribofuranosyl)-6- ethoxy-8-methanesulfonyl-purine (9) was treated with sodium salt of ethyl malonate to give the cyclized nucleosides (4 and 10) in good yields, respectively. Subsequent decarboxylation and deblocking of 4 and 10 afforded 1 and 2 in crystalline form, respectively.