Convenient and Efficient Approach to Di- and Trideoxyribonucleotide N3′→P5′ Phosphoramidates

Abstract

Arbuzov reaction of phenyl phosphorodichloridite with an equivalent of allyl alcohol and tert-butyl alcohol produced ­phenyl allyl H-phosphonate and the subsequent ester-exchange with various nucleosides produced nucleoside allyl H-phosphonates. Antherton-Todd reaction of the nucleoside allyl H-phosphonates with 3′-amino-3′-deoxythymidine yielded dideoxyribonucleotide N3′→P5′ phosphoramidates, and the repetition of this procedure provided trideoxyribonucleotide N3′→P5′ phosphoramidates. The method can be used for the synthesis of oligodeoxyribonucleotide N3′→P5′ phosphoramidates without any protection for all nucleosides.

nucleotide N3′→P5′ phosphoramidate - H-phosphonate - ester-exchange - Antherton-Todd reaction

References

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Spectral data for the representative products 6b and 8b.
Compound 6b: ³¹P NMR (121.5 MHz, DMSO-d ₆): δ = 10.37, 10.26 ppm. ¹H NMR (300 MHz, DMSO-d ₆): δ = 1.76 (s, 3 H, 5-CH₃ of 3′-amino-3′-deoxythymidine), 1.78 (s, 3 H, 5-CH₃ of 3′-deoxythymidine), 2.11-2.17 (m, 5 H, 2′-H of 3′-amino-3′-deoxythymidine and 3′-deoxythymidine and 4′-H of 3′-amino-3′-deoxythymidine), 3.50-3.65 (m, 2 H, 3′-H of 3′-amino-3′-deoxythymidine and 3′-H of 3′-deoxythymidine), 3.65-3.93 (m, 2 H, 5′-H of 3′-amino-3′-deoxythymidine), 4.01 (m, 1 H, 4′-H of 3′-deoxythymidine), 4.01-4.11 (m, 2 H, 5′-H of 3′-deoxythymidine), 4.21-4.30 (m, 1 H, 3′-NH of 3′-amino-3′-deoxythymidine), 4.39-4.47 (m, 2 H, OCH ₂-CH=CH₂), 5.17-5.35 (m, 2 H, OCH₂-CH=CH ₂), 5.88-6.00 (m, 1 H, OCH₂-CH=CH₂), 6.12 (t, 1 H, ³ J = 6.18 Hz, 1′-H of 3′-amino-3′-deoxythymidine), 6.19 (t, 1 H, ³ J = 6.87 Hz, 1′-H of 3′-deoxythymidine), 7.52 (s, 1 H, 6-H of 3′-amino-3′-deoxythymidine), 7.73 (s, 1 H, 6-H of 3′-deoxythymidine), 11.29 (s, 1 H, 3-H of 3′-amino-3′-deoxythymidine), 11.32 (s, 1 H, 3-H of 3′-deoxythymidine) ppm. ¹³C NMR (75 MHz, DMSO-d ₆): δ = 12.11, 12.26, 60.39, 65.58, 65.97, 66.05, 70.21, 70.30, 83.24, 83.88, 84.69, 85.80, 109.21, 109.83, 117.04, 133.53, 133.62, 135.81, 136.18, 150.35, 150.42, 163.70, 163.76 ppm. ESI-MS: m/z = 586.4 [M + H]⁺, 608.2 [M + Na]⁺.
Compound 8b: ³¹P NMR (121.5 MHz, DMSO-d ₆): δ = 10.08, 10.30 ppm. ¹H NMR (300 MHz, DMSO-d ₆): δ = 1.76 (s, 3 H, 5-CH₃ of 3′-amino-3′-deoxythymidine), 1.78 (s, 6 H, 5-CH₃ of 3′-deoxythymidine and 3′-amino-3′-deoxythymidine), 1.99-2.35 (m, 6 H, 6 × H-2′), 3.54-3.86 (m, 5 H, H-4′ of 3′-deoxythymidine and 2 × H-5′), 3.87-4.33 (m, 8 H, 2 × H-4′, 4 × H-5′ and 2 × H-3′ of 3′-amino-3′-deoxythymidine), 4.33-4.50 (m, 3 H, 2 OCH ₂-CH=CH₂ and H-3′ of 3′-deoxythymidine), 5.10-5.38 (m, 4 H, 2 OCH₂-CH=CH ₂), 5.83-6.02 (m, 2 H, 2 OCH₂-CH=CH₂), 6.04-6.26 (m, 3 H, H-1′), 7.54 (s, 2 H, H-6 of 3′-deoxythymidine and 3′-amino-3′-deoxythymidine), 7.74 (s, 1 H, H-6 of 3′-amino-3′-deoxythymidine), 11.23-11.36 (m, 3 H, 3 × H of NH-3) ppm. ¹³C NMR (75 MHz, DMSO-d ₆): δ = 12.60, 12.73, 50.82, 51.22, 60.76, 65.61, 65.76, 66.21, 66.51, 66.57, 66.74, 66.80, 70.59, 70.61, 79.75, 83.72, 83.97, 84.40, 85.03, 85.10, 86.35, 86.49, 109.72, 110.25, 110.34, 117.57, 117.74, 133.90, 133.97, 136.30, 136.72, 150.75, 150.84, 164.18 ppm. ESI-MS: m/z = 951.9 [M + Na]⁺.