An Efficient Protection-Free One-Pot Chemical Synthesis of Modified Nucleoside-5′-Triphosphates

Abstract

A simple, reliable, and an efficient "one-pot, three step" chemical method for the synthesis of modified nucleoside triphosphates such as 5-methylcytidine-5'-triphosphate (5-MeCTP), pseudouridine-5'-triphosphate (pseudoUTP) and N(1)-methylpseudouridine-5'-triphosphate (N(1)-methylpseudoUTP) starting from the corresponding nucleoside is described. The overall reaction involves the monophosphorylation of nucleoside, followed by the reaction with pyrophosphate and subsequent hydrolysis of the cyclic intermediate to furnish the corresponding NTP in moderate yields with high purity (>99.5%).

Recent Trends in Nucleotide Synthesis

“…209−212 It is important to note that the monophosphorylation reaction of ribonucleosides does not require a base in the reaction medium, unlike the successful monophosphorylation reaction of a deoxyribonucleoside, which requires a carefully chosen base. 213 6.1.2. Synthesis via a Phosphoramidate Intermediate.…”

“…Under these optimized conditions, the five natural dNTPs were prepared in 65–70% yields. This protocol was successfully exemplified on base-modified dNTPs. − It is important to note that the monophosphorylation reaction of ribonucleosides does not require a base in the reaction medium, unlike the successful monophosphorylation reaction of a deoxyribonucleoside, which requires a carefully chosen base …”

A selective and atom-economic rearrangement of uridine by cascade biocatalysis for production of pseudouridine

“…Here, expedient access to the Ψ β-C-glycoside core structure represents the principal challenge. Methylation at N1 and conversion into the 5′-triphosphates proceed by transformations well implemented for process scale use [15][16][17][18][19] . Since the first chemical synthesis of Ψ in 1961 20 , the synthetic…”

An Efficient Gram‐Scale Chemical Synthesis of Purine Locked Nucleic Acid Nucleoside‐5′‐O‐Triphosphates

“…Although the use of the “one‐pot, three‐step” strategy for the synthesis of nucleoside triphosphate does not require prior protection and subsequent deprotection of nucleobase and sugar moiety, this strategy often results in poor selectivity and moreover is not applicable to all modified nucleotides (Gillerman & Fischer, 2010). We reported an efficient and improved “one‐pot, three‐step” Ludwig synthetic strategy for the synthesis of deoxynucleoside triphosphates (Kore, Shanmugasundaram, Senthilvelan, & Srinivasan, 2012), modified ribonucleoside triphosphates (Shanmugasundaram, Senthilvelan, Xiao, & Kore, 2016), and inosine triphosphate (Senthilvelan, Shanmugasundaram, & Kore, 2020). It is noteworthy that this improved procedure results in the isolation of the final nucleoside triphosphate with high purity that was optimized by several factors such as amounts of phosphorus oxychloride and pyrophosphate, reaction time, reaction temperature, and proper choice of solvent for the triphosphorylation reaction.…”