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Identification of metabolically stable 5'-phosphate analogs that support single-stranded siRNA activity.

Prakash TP, Lima WF, Murray HM, Li W, Kinberger GA, Chappell AE, Gaus H, Seth PP, Bhat B, Crooke ST, Swayze EE - Nucleic Acids Res. (2015)

Bottom Line: The ss-siRNA activity in vivo requires a metabolically stable 5'-phosphate analog.Our results indicate that the electronic and spatial orientation of the 5'-phosphate analog was critical for ss-siRNA activity.Our study provides guidance to develop surrogate phosphate analog for ss-siRNA and demonstrates that ss-siRNA provides an alternative strategy for therapeutic gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Isis Pharmaceuticals Inc., 2855 Gazelle Ct, Carlsbad, CA 92010, USA tprakash@isisph.com.

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Synthesis of 5′-deoxy-(E)-5′-(dimethylvinylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54.
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Figure 24: Synthesis of 5′-deoxy-(E)-5′-(dimethylvinylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54.

Mentions: Synthesis of 5′-deoxy-(E)-5′-vinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54 was accomplished according to Scheme 8 (Supplementary data) (28). Similarly 5′-deoxy-5′-(Z)-vinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 55 was also synthesized (Figure 12 and Supplementary data) (28). We also synthesized 5′-deoxy-(E)-5′-fluorovinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 56 (Figure 12 and Supplementary data) (34,35) and 5′-deoxy-(Z)-5′-fluorovinyl(dimethylphosphonate)-2′-O-(2-methyl)-thymidine-3′-phosphoramidite 57 (Figure 12 and Supplementary data) using reported methods (36,37).


Identification of metabolically stable 5'-phosphate analogs that support single-stranded siRNA activity.

Prakash TP, Lima WF, Murray HM, Li W, Kinberger GA, Chappell AE, Gaus H, Seth PP, Bhat B, Crooke ST, Swayze EE - Nucleic Acids Res. (2015)

Synthesis of 5′-deoxy-(E)-5′-(dimethylvinylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4381071&req=5

Figure 24: Synthesis of 5′-deoxy-(E)-5′-(dimethylvinylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54.
Mentions: Synthesis of 5′-deoxy-(E)-5′-vinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 54 was accomplished according to Scheme 8 (Supplementary data) (28). Similarly 5′-deoxy-5′-(Z)-vinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 55 was also synthesized (Figure 12 and Supplementary data) (28). We also synthesized 5′-deoxy-(E)-5′-fluorovinyl(dimethylphosphonate)-2′-O-(2-methoxyethyl)-thymidine-3′-phosphoramidite 56 (Figure 12 and Supplementary data) (34,35) and 5′-deoxy-(Z)-5′-fluorovinyl(dimethylphosphonate)-2′-O-(2-methyl)-thymidine-3′-phosphoramidite 57 (Figure 12 and Supplementary data) using reported methods (36,37).

Bottom Line: The ss-siRNA activity in vivo requires a metabolically stable 5'-phosphate analog.Our results indicate that the electronic and spatial orientation of the 5'-phosphate analog was critical for ss-siRNA activity.Our study provides guidance to develop surrogate phosphate analog for ss-siRNA and demonstrates that ss-siRNA provides an alternative strategy for therapeutic gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Isis Pharmaceuticals Inc., 2855 Gazelle Ct, Carlsbad, CA 92010, USA tprakash@isisph.com.

Show MeSH