Synthesis and Stability of a 2′-O-[N-(Aminoethyl)carbamoyl]methyladenosine-Containing Dinucleotide

• Published in: European journal of organic chemistry 2013-11, Vol.2013 (31), p.7184-7192
• Main Authors: Milton, Stefan, Ander, Charlotte, Honcharenko, Dmytro, Honcharenko, Małgorzata, Yeheskiely, Esther, Strömberg, Roger
• Format: Article
• Language: English
• Subjects: Adenosine; Alkylation; Ammonia; Ammonolysis; Ethylenediamine; Hydrolases; Hydrolysis; Läkemedelskemi; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Nucleosides; Nucleotides; Oligonucleotides; Phosphonates; Stability; Synthesis
• ISSN: 1434-193X; 1099-0690
• DOI: 10.1002/ejoc.201300699

Working towards the synthesis of 2′‐O‐[N‐(aminoethyl)carbamoyl]methyl‐modified di‐ and oligonucleotides, we have synthesised a protected 2′‐O‐[N‐(aminoethyl)carbamoyl]methyl‐modified adenosine where the modification is introduced in a convenient one‐pot three‐step procedure. The corresponding H‐phosphonate building block was also synthesised, and from this intermediate, a 2′‐O‐[N‐(aminoethyl)carbamoyl]methyl‐containing dinucleotide could be made. We also performed studies on the chemical and enzymatic stability of this dinucleotide. The dinucleotide was subjected to different ammonolysis and other basic conditions, and HPLC analysis showed that the modification was intact to most conditions, but that there was some minor hydrolysis when NH3 (concd. aq.) was used at 55 °C. Under several other sets of conditions, including saturated NH3 in methanol, and ethylenediamine, the amide remained intact. Treatment of the dinucleotide with Phosphodiesterase I from Crotalus adamanteus venom and Phosphodiesterase II from bovine spleen showed that the N‐(aminoethyl)carbamoylmethyl moiety gives the phosphodiester linkage substantial protection against enzymatic degradation; the phosphodiester was not degraded by PDE II at all after seven days. A 2′‐O‐[N‐(aminoethyl)carbamoyl]methyl‐modified adenosine and a corresponding dinucleotide were synthesised. Hydrolysis (1–2 %) was observed in concentrated aqueous NH3 at 55 °C, but under several other sets of reaction conditions, the amide remained intact. The N‐(aminoethyl)carbamoylmethyl moiety gave substantial protection against enzymatic degradation by nucleases from snake venom and bovine spleen.