Rapid Synthesis of Nucleoside Triphosphates and Analogues

摘要

Nucleoside triphosphates (NTPs) are essential biomolecules involved in almost all biological processes, and their study is therefore critical to understanding cellular biology. Here, we describe a chemical synthesis suitable for obtaining both naturaland highly modified NTPs, which can, for example, be used as surrogates to probe biological processes. The approach includes the preparation of a reagent that enables the facile introduction and modification of three phosphate units: cyclic pyrophosphoryl P-amidite (c-PyPA), derived from pyrophosphate (PV) and a reactive phosphoramidite (Pm). By using non-hydrolyzable analogues of pyrophosphate, the reagent can be readily modified to obtain a family of non-hydrolyzable analogues containing CH_2, CF_ 2,CCl_2, and NH that are stable in solution for several weeks if stored appropriately. They enable the synthesis of NTPs by reaction with nucleosides to give deoxycyclotriphosphate esters that are then oxidized to cyclotriphosphate (cyclo-TP) esters. Theuse of different oxidizing agents provides an opportunity for modification at P-a. Furthermore, terminal modifications at P-y can be introduced by linearization of the cyclo-TP ester with various nucleophiles.