Atsumi Naoko, Ueno Yoshihito, Kanazaki Makiko, Shuto Satoshi, Matsuda Akira
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Bioorg Med Chem. 2002 Sep;10(9):2933-9. doi: 10.1016/s0968-0896(02)00141-4.
In order to develop novel antigene molecules forming thermally stable triplexes with target DNAs and having nuclease resistance properties, we synthesized oligodeoxynucleotides (ODNs) with various lengths of aminoalkyl-linkers at the 4'alpha position of thymidine and the aminoethyl-linker at the 4'alpha position of 2'-deoxy-5-methylcytidine. Thermal stability of triplexes between these ODNs and a DNA duplex was studied by thermal denaturation. The ODNs containing the nucleoside 2 with the aminoethyl-linker or the nucleoside 3 with the aminopropyl-linker thermally stabilized the triplexes, whereas the ODNs containing the nucleoside 1 with the aminomethyl-linker or the nucleoside 4 with the 2-[N-(2-aminoethyl)carbamoyl]oxy]ethyl-linker thermally destabilized the triplexes. The ODNs containing 2 were the most efficient at stabilizing the triplexes with the target DNA. The ODNs containing 4'alpha-C-(2-aminoethyl)-2'-deoxy-5-methylcytidine (5) also efficiently stabilized the triplexes with the target DNA. Stability of the ODN containing 5 to nucleolytic hydrolysis by snake venom phosphodiesterase (a 3'-exonuclease) was studied. It was found that the ODN containing 5 was more resistant to nucleolytic digestion by the enzyme than an unmodified ODN. In a previous paper, we reported that the ODNs containing 2 were more resistant to nucleolytic digestion by DNase I (an endonuclease) than the unmodified ODNs. Thus, it was found that the ODNs containing 4'alpha-C-(2-aminoethyl)-2'-deoxynucleosides were good candidates for antigene molecules.
为了开发能与靶DNA形成热稳定三链体且具有核酸酶抗性的新型反基因分子,我们在胸苷的4'α位合成了带有不同长度氨基烷基连接子的寡脱氧核苷酸(ODN),并在2'-脱氧-5-甲基胞苷的4'α位合成了氨乙基连接子。通过热变性研究了这些ODN与DNA双链体之间三链体的热稳定性。含有带有氨乙基连接子的核苷2或带有氨丙基连接子的核苷3的ODN使三链体热稳定,而含有带有氨甲基连接子的核苷1或带有2-[N-(2-氨乙基)氨基甲酰氧基]乙基连接子的核苷4的ODN使三链体热不稳定。含有2的ODN在稳定与靶DNA的三链体方面效率最高。含有4'α-C-(2-氨乙基)-2'-脱氧-5-甲基胞苷(5)的ODN也能有效地稳定与靶DNA的三链体。研究了含有5的ODN对蛇毒磷酸二酯酶(一种3'-外切核酸酶)核酸水解的稳定性。发现含有5的ODN比未修饰的ODN对该酶的核酸消化更具抗性。在之前的一篇论文中,我们报道含有2的ODN比未修饰的ODN对DNase I(一种内切核酸酶)的核酸消化更具抗性。因此,发现含有4'α-C-(2-氨乙基)-2'-脱氧核苷的ODN是反基因分子的良好候选物。