Aqueous Heck Cross-Coupling Preparation of Acrylate-Modified Nucleotides and Nucleoside Triphosphates for Polymerase Synthesis of Acrylate-Labeled DNA

• Published in: Journal of organic chemistry 2013-10, Vol.78 (19), p.9627-9637
• Main Authors: Dadová, Jitka, Vidláková, Pavlína, Pohl, Radek, Havran, Luděk, Fojta, Miroslav, Hocek, Michal
• Format: Article
• Language: English
• Subjects: Acrylates - chemical synthesis; Acrylates - chemistry; Adenine - analogs & derivatives; Adenine - chemical synthesis; Adenine - chemistry; DNA-Directed DNA Polymerase - chemistry; DNA-Directed DNA Polymerase - metabolism; Guanine - analogs & derivatives; Guanine - chemical synthesis; Guanine - chemistry; Molecular Structure; Nucleosides - chemical synthesis; Nucleosides - chemistry; Nucleotides - chemical synthesis; Nucleotides - chemistry
• ISSN: 0022-3263; 1520-6904
• DOI: 10.1021/jo4011574

Aqueous-phase Heck coupling methodology was developed for direct attachment of butyl acrylate to 5-iodoracil, 5-iodocytosine, 7-iodo-7-deazaadenine, and 7-iodo-7-deazaguanine 2′-deoxyribonucleoside 5′-O-monophosphates (dNMPs) and 5′-O-triphosphates (dNTPs) and compared with the classical approach of phosphorylation of the corresponding modified nucleosides. The 7-substituted 7-deazapurine nucleotides (dA BA MP, dA BA TP, dG BA MP, and dG BA TP) were prepared by the direct Heck coupling of nucleotides in good yields (35–55%), whereas the pyrimidine nucleotides reacted poorly and the corresponding BA-modified dNTPs were prepared by triphosphorylation of the modified nucleosides. The acrylate-modified dN BA TPs (N = A, C, and U) were good substrates for DNA polymerases and were used for enzymatic synthesis of acrylate-modified DNA by primer extension, whereas dG BA TP was an inhibitor of polymerases. The butyl acrylate group was found to be a useful redox label giving a strong reduction peak at −1.3 to −1.4 V in cyclic voltammetry.