A Study on Synthesis and Upscaling of 2'- O-AECM-5-methyl Pyrimidine Phosphoramidites for Oligonucleotide Synthesis

Molecules. 2021 Nov 17;26(22):6927. doi: 10.3390/molecules26226927.

Abstract

2'-O-(N-(Aminoethyl)carbamoyl)methyl-modified 5-methyluridine (AECM-MeU) and 5-methylcytidine (AECM-MeC) phosphoramidites are reported for the first time and prepared in multigram quantities. The syntheses of AECM-MeU and AECM-MeC nucleosides are designed for larger scales (approx. 20 g up until phosphoramidite preparation steps) using low-cost reagents and minimizing chromatographic purifications. Several steps were screened for best conditions, focusing on the most crucial steps such as N3 and/or 2'-OH alkylations, which were improved for larger scale synthesis using phase transfer catalysis (PTC). Moreover, the need of chromatographic purifications was substantially reduced by employing one-pot synthesis and improved work-up strategies.

Keywords: 2′-O-(N-(aminoethyl)carbamoyl)methyl modification; 5-methylcytidine; 5-methyluridine; alkylation; monoacetylation; oligonucleotides; phase transfer catalysis (PTC).

MeSH terms

  • Cytidine / analogs & derivatives*
  • Cytidine / chemistry
  • Oligonucleotides / chemical synthesis*
  • Oligonucleotides / chemistry*
  • Organophosphorus Compounds / chemistry*
  • Uridine / analogs & derivatives*
  • Uridine / chemistry

Substances

  • Oligonucleotides
  • Organophosphorus Compounds
  • phosphoramidite
  • ribothymidine
  • Cytidine
  • 5-methylcytidine
  • Uridine

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