Bookser Brett C, Raffaele Nicholas B
Metabasis Therapeutics, Inc., 11119 North Torrey Pines Road, La Jolla, California 92037, USA.
J Comb Chem. 2008 Jul-Aug;10(4):567-72. doi: 10.1021/cc8000212. Epub 2008 May 29.
A high-throughput phosphoramidite method for HepDirect prodrug synthesis was optimized on seven representative nucleosides, adenosine, inosine, guanosine, uridine, cytidine, AICA-riboside, and thymidine, each on a 5 mg scale. The variables optimized included (1) reaction time, (2) reaction temperature, (3) activating agent, (4) solvent, (5) purification method, and (6) stoichiometry. Preparative HPLC with mass-based fraction collection and yield determination from an ELSD standard curve enabled high-throughput. The optimized conditions for the representative nucleosides required 6 mol equiv of phosphoramidite to nucleoside and resulted in an average HPLC determined yield of 31 +/- 14% and HPLC purity of 93 +/- 3%.
一种用于HepDirect前药合成的高通量亚磷酰胺方法在七种代表性核苷（腺苷、肌苷、鸟苷、尿苷、胞苷、氨基咪唑甲酰胺核苷和胸苷）上进行了优化，每种核苷的规模为5毫克。优化的变量包括：（1）反应时间，（2）反应温度，（3）活化剂，（4）溶剂，（5）纯化方法，以及（6）化学计量比。采用基于质量的馏分收集和蒸发光散射检测器（ELSD）标准曲线进行产率测定的制备型高效液相色谱（HPLC）实现了高通量。代表性核苷的优化条件需要核苷与亚磷酰胺的摩尔比为6:1，HPLC测定的平均产率为31±14%，HPLC纯度为93±3%。
