title: Gram-Scale Chemical Synthesis of Base-Modified Ribonucleoside-5'-O-Triphosphates.
abstract: This unit delineates a simple, reliable, straight-forward, general, and efficient chemical method for the synthesis of modified nucleoside-5'-O-triphosphates such as 5-methylcytidine-5'-O-triphosphate (5-Me-CTP), pseudouridine-5'-O-triphosphate (pseudo-UTP), and N1-methylpseudouridine-5'-O-triphosphate (N1-methylpseudo-UTP), starting from the corresponding nucleoside. The reaction utilizes an improved protection-free "one-pot, three-step" Ludwig synthetic strategy that involves the monophosphorylation of the nucleoside with phosphorous oxychloride followed by reaction with tributylammonium pyrophosphate and subsequent hydrolysis of the resulting cyclic intermediate to furnish the corresponding ribonucleoside triphosphate (NTP) in moderate yields. It is noteworthy that the reaction affords high purity (>99.5%) NTPs after DEAE Sepharose column purification. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Life Sciences Solutions Group, Thermo Fisher Scientific, Austin, Texas.
authors.forename: Muthian
authors.initials: M
authors.lastname: Shanmugasundaram
authors.affiliation: Life Sciences Solutions Group, Thermo Fisher Scientific, Austin, Texas.
authors.forename: Annamalai
authors.initials: A
authors.lastname: Senthilvelan
authors.affiliation: Life Sciences Solutions Group, Thermo Fisher Scientific, Austin, Texas.
authors.forename: Anilkumar R
authors.initials: AR
authors.lastname: Kore
mesh_terms: D010766:Phosphorylation; D012263:Ribonucleosides / Q000138:chemical synthesis*
chemical_list: D012263:Ribonucleosides
keywords: chemical synthesis; gene therapy; messenger RNA; one-pot synthesis; ribonucleotides; vaccination therapy