含亚甲基缩醛键的寡脱氧核苷酸的合成及杂交性质的初步研究  

Synthesis and hybridizing properties of oligodeoxynucleotide analogs containing methyleneformacetal

作　　者：余建鑫[1] 张万年[1] 张蕾[1] 周有骏[1] 吕加国[1] 

机构地区：[1]第二军医大学药学院,上海200433

出　　处：《药学学报》2003年第1期27-31,共5页Acta Pharmaceutica Sinica

摘　　要：目的　本文设计合成了一类新型混合骨架的寡脱氧核苷酸 (MBO)。方法　 3′ O (二苯膦酰氧 )甲基缩醛核苷 (1)在三甲基硅三氟甲磺酸酯 (TMSOTf)条件下 ,与 3′ 位保护的脱氧核苷 (2或 6 )缩合。得到的二聚 (3)或三聚体 (7)的 5′ 位经 4 ,4′ 二甲氧三苯甲基 (DMTr)保护 ,3′ 位与 (2 氰乙基N ,N 二异丙基 )氯化亚磷酰胺缩合 ,然后应用标准固相DNA合成法掺入到寡核苷酸中。结果　本文合成了 6条带亚甲基缩醛键的寡核苷酸 (ODN II～ODN VII) ,考察了它们的杂交性质 ,测定了与互补DNA的解链温度Tm 值。结论　此类寡核苷酸平均每个亚甲基缩醛键的修饰 ,解链温度Tm 值下降约 0 8～ 1 2℃ ,杂交亲和力与对照的天然磷酸二酯键的寡核苷酸相当。Aim To design and synthesize a new mixed backbone oligonucleotide (MBO). Methods In the presence of trimethylsilyl trifluoromethane-sulfonate (TMSOTf), condensation of 3′- O -(diphenylphosphinyloxy) methyl acetal (1) with 3′-protected deoxynucleoside (2 or 6) afforded dimers (3) or trimers (7) respectively. 5′-Hydroxyl and 3′-hydroxyl groups of these acetal-linked oligomers were protected by 4,4′-dimethoxytriphenylmethyl (DMTr) or by diisopropylamino-β-cyanothoxyphosphine respectively. Then, compounds 5 and 9) were incorporated into oligonucleotides by using the standard solid-phase synthesis of DNA with the phosphoramidite method. Results Six new oligonucleotides (ODN-II-ODN-VII) containing methyleneformacetal have been synthesized. The melting temperatures ( T m ) of these ODNs with their DNA complements were determined. Conclusion The melting temperatures ( T m ) of these modified ODNs were lowered about 0.8-1.2 ℃ per methyleneformacetal modification. These new ODNs can hybridize to DNA with only slightly less affinity than a control phosphodiester ODN, yet more work is necessary to study these modified ODNs and their biological activities.

关 键 词：反义药物 寡脱氧核苷酸 解链温度 杂交 MBO 亚甲基缩醛键 

分 类 号：R914.4[医药卫生—药物化学]