PEG-OLIGODEOXYRIBONUCLEOTIDE CONJUGATES

PEG-寡脱氧核糖核苷酸缀合物

• 批准号: 3770389
• 负责人: S L BEAUCAGE
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3770389
• 关键词: chemical conjugate; chemical synthesis; ethylene glycols; human immunodeficiency virus; nucleic acid hybridization; nucleotide analog; oligonucleotides; polyethylenes; protein structure; virus replication

Tailing oligonucleotide analogues with hydrophobic molecules may facilitate the entry of the oligonucleotide conjugates into living cells and, thereby, improve the potency of antisense molecules in inhibiting gene expression. However, the size of the PEG conjugated to oligonucleotides may adversely affect the hybridization properties of the conjugates. An in vitro assay has been developed to evaluate the hybridization abilities of PEG-oligonucleotide conjugates with a complementary HIV-TAR RNA oligomer (250 nt.). The synthesis of oligodeoxyribonucleotides using a solid support derivatized with PEG (M.W.: 300, 2,000 or 5,000) has unexpectedly been facile. Conversely, the condensation of activated PEG-derivatives with the 5'-end of an oligonucleotide anchored to a solid support has been difficult. In this regard, significant progress has, however, been achieved through the use of solid-phase bound oligonucleotides derivatized with novel nucleophilic linkers. Although, the hybridization of PEG-oligonucleotide conjugates with complementary DNA sequences having equal number of nucleobases was not affected by the size of the PEG tail, PEG-oligonucleotide conjugates did not hybridize as well with the 250 bases long RNA oligomer. Digestion of hybrids composed of PEG-oligonucleotide conjugates and radiolabelled HIV- TAR RNA with ribonuclease A and ribonuclease T1 showed that the oligonucleotide conjugated with the PEG (M.W. 5,000) did not significantly hybridize with the RNA relative to an unmodified DNA oligomer. While the oligonucleotide conjugated with PEG (M.W. 300) hybridized as well as the parent oligonucleotide with the target RNA, the oligonucleotide conjugated to PEG (M.W. 2,000) hybridized to the extent of ca. 50% with HIV-TAR RNA under the same conditions. The above assay did not, however, provide evidence indicating that the PEG portion of the conjugate could have wrapped around or entangled with the RNA as a means to further suppress translation. It is, nevertheless likely that PEG should protect the oligonucleotide moiety of the conjugates against exonucleases. The potency of an alpha,beta-oligodeoxyribonucleotide conjugated to PEG (M.W. 300) toward the inhibition of HIV replication in a chronically infected human T-cell line is being evaluated.

具有疏水分子的加尾寡核苷酸类似物可 促进寡核苷酸缀合物进入活细胞 从而提高反义分子抑制肿瘤生长的效力 基因表达。 然而，缀合至PEG的PEG的大小取决于PEG的分子量。 寡核苷酸可能不利地影响寡核苷酸的杂交性质。 结合物。 已经开发了一种体外试验来评估 PEG-寡核苷酸偶联物与 互补HIV-TAR RNA寡聚体（250 nt.）。 固相载体法合成寡脱氧核苷酸 用PEG衍生化（M.W.：300、2,000或5,000）， 很容易 相反，活化的PEG衍生物与 锚定在固体支持物上的寡核苷酸的5 ′-末端已经 难 然而，在这方面取得了重大进展， 通过使用固相结合的寡核苷酸衍生 与新型亲核连接体的反应。 尽管PEG-寡核苷酸缀合物与 具有相等数目的核碱基的互补DNA序列不被认为是 受PEG尾部大小的影响，PEG-寡核苷酸缀合物确实 不与250个碱基长的RNA寡聚体杂交。 消化 由PEG-寡核苷酸缀合物和放射性标记的HIV-1组成的杂交体， TAR RNA与核糖核酸酶A和核糖核酸酶T1的结合表明， 与PEG缀合的寡核苷酸（M.W. 5,000）没有显著 与相对于未修饰的DNA寡聚体的RNA杂交。 而 与PEG缀合的寡核苷酸（M.W. 300)杂交以及 亲本寡核苷酸与靶RNA，寡核苷酸缀合 至PEG（M.W. 2,000）杂交到CA的程度。50%携带HIV-TAR RNA 在相同的条件下。 然而，上述测定没有提供 有证据表明，缀合物的PEG部分可能具有 包裹或缠绕在RNA上，作为进一步抑制 翻译. 然而，PEG可能会保护 在一些实施方案中，缀合物的寡核苷酸部分针对核酸外切酶。 的 与PEG缀合的α，β-寡脱氧核糖核苷酸的效力（M.W. 300)在慢性感染者中抑制HIV复制 人类T细胞系正在评估中。