The present disclosure provides a solid phase method of making oligonucleotides via sequential coupling cycles including at least one coupling of a dinucleotide dimer subunit to a free 3'-terminal group of a growing chain. The oligonucleotides include at least two nucleoside subunits joined by a N3'→P5' phosphoramidate linkage. The method may include the steps of (a) deprotecting the protected 3' amino group of a terminal nucleoside attached to a solid phase support, said deprotecting forming a free 3' amino group; (b) contacting the free 3' amino group with a 3'-protected amino-dinucleotide-5'-phosphoramidite dimer in the presence of a nucleophilic catalyst to form an internucleoside N3'→P5' phosphoramidite linkage; and (c) oxidizing (e.g., sulfurizing) the linkage. The compositions produced by the subject methods may include a reduced amount of one or more (N-x) oligonucleotide products. Also provided are pharmaceutical compositions including the subject oligonucleotide compositions.
本公开提供了一种通过顺序偶联循环制备寡核苷酸的固相方法,其中至少包括将二核苷酸二聚体亚基偶联到生长链的自由3'-末端基团的至少一个偶联步骤。这些寡核苷酸包括至少两个核苷酸亚基,通过N3'→P5'
磷酰胺酸酯键连接。该方法可能包括以下步骤:(a)去保护连接到固相支持物上的末端核苷酸的保护3'
氨基基团,去保护形成自由的3'
氨基基团;(b)在存在亲核催化剂的情况下,将自由的3'
氨基基团与3'-保护
氨基-二核苷酸-5'-
磷酰胺酯二聚体反应,形成核苷酸之间的N3'→P5'
磷酰胺酯键;(c)氧化(例如,
硫化)该键。通过该方法生产的组合物可能包括减少一种或多种(N-x)寡核苷酸产物的数量。还提供了包括该寡核苷酸组合物的制药组合物。