Laboratory of Medicinal Chemistry, Department of Chemistry, Clemson University, Clemson, SC 29634, United States.
Bioorg Med Chem Lett. 2011 Aug 15;21(16):4788-92. doi: 10.1016/j.bmcl.2011.06.058. Epub 2011 Jun 21.
A series of neomycin dimers have been synthesized using 'click chemistry' with varying linker functionality and length to target the TAR RNA region of HIV virus. TAR (trans activation response) RNA region, a 59 base pair stem loop structure located at 5'-end of all nascent HIV-1 transcripts interacts with a key regulatory protein, Tat, and necessitates the replication of HIV-1 virus. Neomycin, an aminosugar, has been shown to exhibit more than one binding site with HIV TAR RNA. Multiple TAR binding sites of neomycin prompted us to design and synthesize a small library of neomycin dimers using click chemistry. The binding between neomycin dimers and HIV TAR RNA was characterized using spectroscopic techniques including FID (Fluorescent Intercalator Displacement) titration and UV-thermal denaturation. UV thermal denaturation studies demonstrate that neomycin dimer binding increase the melting temperature (T(m)) of the HIV TAR RNA up to 10°C. Ethidium bromide displacement titrations revealed nanomolar IC(50) between neomycin dimers and HIV TAR RNA, whereas with neomycin, a much higher IC(50) in the micromolar range is observed.
已使用“点击化学”合成了一系列新霉素二聚体，这些二聚体具有不同的连接子功能和长度，旨在靶向 HIV 病毒的 TAR RNA 区域。TAR（转录激活反应）RNA 区域是位于所有新生 HIV-1 转录本 5'端的 59 个碱基对茎环结构，与关键调节蛋白 Tat 相互作用，是 HIV-1 病毒复制所必需的。新霉素是一种氨基糖，已被证明与 HIV TAR RNA 具有不止一个结合位点。新霉素的多个 TAR 结合位点促使我们使用点击化学设计和合成了一小部分新霉素二聚体文库。使用包括 FID（荧光嵌入剂置换）滴定和 UV-热变性在内的光谱技术来表征新霉素二聚体与 HIV TAR RNA 之间的结合。UV 热变性研究表明，新霉素二聚体的结合使 HIV TAR RNA 的熔点（T(m)）升高了 10°C。溴化乙锭置换滴定显示新霉素二聚体与 HIV TAR RNA 之间的纳摩尔 IC(50)，而新霉素则观察到更高的微摩尔范围的 IC(50)。
