Here we report on the design, preparation and investigation of four analogues of the anti-HIV G-quadruplex-forming Hotoda’s aptamer, based on an unprecedented linear topology. In these derivatives, four TGGGAGT tracts have been joined together by exploiting 3′-3′ and 5′-5′ inversion of polarity sites formed by canonical phosphodiester bonds or a glycerol-based linker. Circular dichroism data suggest that all oligodeoxynucleotides fold in monomolecular G-quadruplex structures characterized by a parallel strand orientation and three side loops connecting 3′- or 5′-ends. The derivative bearing two lipophilic groups, namely HT353LGly, inhibited virus entry into the host cell, with anti-HIV-1 activity in the low nanomolar range; the other derivatives, albeit sharing the same base sequence and similar topology, were inactive. These results highlight that monomolecular Hotoda’s aptamers with inversion of polarity sites represent a successful alternative strategy that merges the easiness of synthesis with the maintenance of remarkable activity. They also indicate that two lipophilic groups are necessary and sufficient for biological activity. Our data will inspire the design of further simplified derivatives with improved biophysical and antiviral properties.

在这里，我们报告了基于前所未有的线性拓扑结构的抗 HIV G-四链体形成 Hotoda 适体的四种类似物的设计、制备和研究。在这些衍生物中，通过利用由规范磷酸二酯键或基于甘油的接头形成的极性位点的 3'-3' 和 5'-5' 反转，将四个 TGGGAGT 区域连接在一起。圆二色性数据表明，所有寡脱氧核苷酸在单分子 G-四链体结构中折叠，其特征是平行链方向和连接 3'-或 5'-末端的三个侧环。带有两个亲脂基团的衍生物，即 HT353LGly，抑制病毒进入宿主细胞，在低纳摩尔范围内具有抗 HIV-1 活性；其他衍生物虽然共享相同的碱基序列和相似的拓扑结构，但不活跃。这些结果突出表明，具有极性位点反转的单分子 Hotoda 适体代表了一种成功的替代策略，它将合成的容易性与显着活性的维持相结合。他们还表明，两个亲脂基团对于生物活性是必要的和足够的。我们的数据将激发具有改进的生物物理和抗病毒特性的进一步简化衍生物的设计。