Convenient synthesis of nucleoside 5'-(α-P-thio)triphosphates and phosphorothioate nucleic acids(DNA and RNA)  

作　　者：CATON-WILLIAMS Julianne FIAZ Bilal HOXHAJ Rudiona SMITH Matthew 

机构地区：[1]Department of Chemistry,Georgia State University

出　　处：《Science China Chemistry》2012年第1期80-89,共10页中国科学（化学英文版）

基　　金：supported by USA NIH(GM095086);the Georgia Cancer Coalition(GCC) Distinguished Cancer Clinicians and Scientists;USA National Science Foundation(MCB-0824837)

摘　　要：Modified deoxy-and ribo-nucleoside triphosphates are chemically synthesized in multiple steps due to the protection and deprotection of the nucleoside functionalities.To conveniently synthesize the S-modified triphosphates for enzymatically preparing phosphorothioate DNAs and RNAs(PS-DNA and PS-RNA) as potential therapeutics,herein we report a one-pot strategy to synthesize the deoxy-and ribo-nucleoside 5'-(α-P-thio)triphosphates(dNTPαS and NTPαS) without protecting any nucleoside functionalities.This facile synthesis is achieved by treating the nucleosides with a mild phosphitylating reagent,reacting selectively with the 5'-hydroxyl group of each unprotected nucleoside,followed by sulfurization and hydrolysis to afford the crude dNTPαS and NTPαS analogs(mixtures of Sp and Rp diastereomers).We also demonstrated that after just simple precipitation(without HPLC and ion-exchange purification),the quality of the synthesized dNTPαS and NTPαS analogs is excellent for direct DNA polymerization and RNA transcription,respectively.Since Klenow DNA polymerase and T7 RNA polymerase accept the Sp diastereomers of dNTPαS and NTPαS analogs,respectively,while the Rp diastereomers are neither substrates nor inhibitors,the diastereomerically-pure PS-DNAs and PS-RNAs can be conveniently synthesized enzymatically.Modified deoxy-and ribo-nucleoside triphosphates are chemically synthesized in multiple steps due to the protection and deprotection of the nucleoside functionalities.To conveniently synthesize the S-modified triphosphates for enzymatically preparing phosphorothioate DNAs and RNAs（PS-DNA and PS-RNA） as potential therapeutics,herein we report a one-pot strategy to synthesize the deoxy-and ribo-nucleoside 5＇-（α-P-thio）triphosphates（dNTPαS and NTPαS） without protecting any nucleoside functionalities.This facile synthesis is achieved by treating the nucleosides with a mild phosphitylating reagent,reacting selectively with the 5＇-hydroxyl group of each unprotected nucleoside,followed by sulfurization and hydrolysis to afford the crude dNTPαS and NTPαS analogs（mixtures of Sp and Rp diastereomers）.We also demonstrated that after just simple precipitation（without HPLC and ion-exchange purification）,the quality of the synthesized dNTPαS and NTPαS analogs is excellent for direct DNA polymerization and RNA transcription,respectively.Since Klenow DNA polymerase and T7 RNA polymerase accept the Sp diastereomers of dNTPαS and NTPαS analogs,respectively,while the Rp diastereomers are neither substrates nor inhibitors,the diastereomerically-pure PS-DNAs and PS-RNAs can be conveniently synthesized enzymatically.

关 键 词：sulfur modification S-derivatized nucleic acid nucleoside triphosphates nucleoside phosphorothioates phosphorothioate DNA and RNA DIASTEREOMERS DNA and RNA polymerases 

分 类 号：O629.7[理学—有机化学]