Abstract
The solution phase synthesis of the tetraribonucleoside triphosphate r(ApCpGpU) 18 and the corresponding cyclic tetraribonucleotide 19 is described. The synthetic methodology is based on 5'- O -(DMTr)-2'- O -(Fpmp)-ribonucleoside-3'- H -phosphonate building blocks 10. Coupling, which is rapid and quantitative, is effected with di-(2-chlorophenyl) phosphorochloridate 5 at -40 degreesC; it is followed by in situ treatment with 2-(4-methyl-phenyl)sulphanyl-1 H -isoindole-1,3(2 H )-dione 6b. The resulting sulphur transfer reaction also proceeds rapidly and quantitatively at -40 degreesC. The same coupling and sulphur transfer steps are used in the cyclization reaction, but a 5'- H -phosphonate intermediate 24 is involved. The final three-step unblocking process involves treatment with (i) E -2-nitrobenzaldoxime 7 and N 1, N 1, N 3, N 3-tetramethylguanidine (TMG) 8 in aceto-nitrile, (ii) concentrated aqueous ammonia at 50 degreesC and (iii) 0.5 mol/dm3sodium acetate buffer (pH 4.0) at 40 degreesC. The fully unblocked products 18 and 19 were characterized by NMR spectroscopy and by enzymatic digestion.
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