Formamide → Uridine 5'-triphosphate

Participants

Reaction Steps

Formamide (2.5 ml) and pyridine (7.6 ml) were added to a dehydrated triethylamine salt (10 mmol) of pyrophosphate (TEA-PPi), followed by stirring the resultant mixture. To a dehydrated tributylamine salt of uridine 5′-monophosphate (UMP-TBA) (10 mmol) placed in another vessel, DMAC (3.6 ml), dioxane (3.2 ml), and tributylamine (3.3 ml) were added and stirred, and DPC (2.3 ml) was added dropwise thereto. The mixture was stirred at room temperature for one hour, to thereby form UMP-DPP, which was added to the aforementioned dehydrated solution of TEA-PPi prepared in advance. The reaction mixture was stirred at room temperature for one hour, forming UTP. DMAC (7.2 ml) was added to a tributylamine salt of 2′-deoxycytidine 5′-monophosphate (TBA-dCMP) (4.9 g, 10 mmol) placed in another vessel, to thereby form a suspension, and DPC (2.2 ml, 1.1 equivalents) was added to the suspension. The resultant mixture was stirred for 40 minutes, and tributylamine (TBA) (9.5 ml) was added to the mixture. Another 20 minutes stirring was carrier out, to thereby prepare dCMP-DPP. The thus-prepared dCMP-DPP solution was added to the aforementioned synthesized UTP solution prepared in advance, and the mixture was stirred at room temperature for 56 hours. Reaction was terminated by adding water, and the pH of the reaction mixture was adjusted to 11 by adding a 30% aqueous sodium hydroxide solution. After the solvent was removed by concentration, the pH was adjusted to 7.0 by adding a 6 mol/L hydrochloric acid. The mixture was partitioned with ethyl acetate. The aqueous layer formed by partitioning was subjected to HPLC analysis (272 nm), revealing that the synthesis yield of dCP4U was 37.7%.

Provenance & Evidence