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High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

Bioconjugate chemistry (2014-11-15)

Daniel Zewge, Francis Gosselin, Denise M Kenski, Jenny Li, Vasant Jadhav, Yu Yuan, Sandhya S Nerurkar, David M Tellers, W Michael Flanagan, Ian W Davies

ABSTRACT

Chemical modification of siRNA is achieved in a high-throughput manner (96-well plate format) by copper catalyzed azide-alkyne cycloadditions. This transformation can be performed in one synthetic operation at up to four positions with complete specificity, good yield, and acceptable purity. As demonstrated here, this approach extends the current synthetic options for oligonucleotide modifications and simultaneously facilitates the systematic, rapid biological evaluation of modified siRNA.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Methanol, NMR reference standard

Sigma-Aldrich

Benzyl azide solution, ~0.5 M in dichloromethane, ≥95.0% (HPLC)

Imidazole, European Pharmacopoeia (EP) Reference Standard

Ondansetron impurity E, European Pharmacopoeia (EP) Reference Standard

Supelco

Residual Solvent - Acetonitrile(solution in DMSO), Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

Triethylamine, for amino acid analysis, ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

Sigma-Aldrich

Water, PCR Reagent

Sigma-Aldrich

Trichloroacetic acid, ≥99.0% (titration)

Sigma-Aldrich

Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))

Supelco

Water, ACS reagent, for ultratrace analysis

Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C

Sigma-Aldrich

Methanol, Laboratory Reagent, ≥99.6%

Sigma-Aldrich

Methanol, suitable for HPLC, ≥99.9%

Sigma-Aldrich

Methanol, HPLC Plus, ≥99.9%

Sigma-Aldrich

Methanol, Absolute - Acetone free

Supelco

2,6-Lutidine, analytical standard

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Methanol, ACS spectrophotometric grade, ≥99.9%

Supelco

Methanol, analytical standard

Sigma-Aldrich

E-Toxate^™ Water, endotoxin, free

Supelco

Water, for HPCE, for luminescence, suitable for UV/Vis spectroscopy

Sigma-Aldrich

Water, Deionized