Hwang Seongwoo, Tamilarasu Natarajan, Kibler Karen, Cao Hong, Ali Akbar, Ping Yueh-Hsin, Jeang Kuan-Teh, Rana Tariq M
Chemical Biology Program, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605-2324, USA.
J Biol Chem. 2003 Oct 3;278(40):39092-103. doi: 10.1074/jbc.M301749200. Epub 2003 Jul 11.
Antiretroviral therapy to treat AIDS uses molecules that target the reverse transcriptase and protease enzymes of human immunodeficiency virus, type 1 (HIV-1). A major problem associated with these treatments, however, is the emergence of drug-resistant strains. Thus, there is a compelling need to find drugs against other viral targets. One such target is the interaction between Tat, an HIV-1 regulatory protein essential for viral replication, and trans-activation-responsive (TAR) RNA. Here we describe the design and synthesis of an encoded combinatorial library containing 39,304 unnatural small molecules. Using a rapid high through-put screening technology, we identified 59 compounds. Structure-activity relationship studies led to the synthesis of 19 compounds that bind TAR RNA with high affinities. In the presence of a representative Tat-TAR inhibitor (5 microM TR87), we observed potent and sustained suppression of HIV replication in cultured cells over 24 days. The same concentration of this inhibitor did not exhibit any toxicity in cell cultures or in mice. TR87 was also shown to specifically disrupt Tat-TAR binding in vitro and inhibit Tat-mediated transcriptional activation in vitro and in vivo, providing a strong correlation between its activities and inhibition of HIV-1 replication. These results provide a structural scaffold for further development of new drugs, alone or in combination with other drugs, for treatment of HIV-1-infected individuals. Our results also suggest a general strategy for discovering pharmacophores targeting RNA structures that are essential in progression of other infectious, inflammatory, and genetic diseases.
用于治疗艾滋病的抗逆转录病毒疗法使用的分子靶向人类免疫缺陷病毒1型（HIV-1）的逆转录酶和蛋白酶。然而，这些治疗方法的一个主要问题是耐药菌株的出现。因此，迫切需要找到针对其他病毒靶点的药物。这样一个靶点是Tat（一种对病毒复制至关重要的HIV-1调节蛋白）与反式激活应答（TAR）RNA之间的相互作用。在此，我们描述了一个包含39304个非天然小分子的编码组合文库的设计与合成。使用快速高通量筛选技术，我们鉴定出了59种化合物。构效关系研究促使合成了19种与TAR RNA具有高亲和力结合的化合物。在一种代表性的Tat-TAR抑制剂（5微摩尔TR87）存在的情况下，我们观察到在培养细胞中HIV复制在24天内受到有效且持续的抑制。相同浓度的这种抑制剂在细胞培养物或小鼠中未表现出任何毒性。TR87还被证明在体外能特异性破坏Tat-TAR结合，并在体外和体内抑制Tat介导的转录激活，这表明其活性与抑制HIV-1复制之间存在很强的相关性。这些结果为进一步开发单独或与其他药物联合用于治疗HIV-1感染个体的新药提供了一个结构框架。我们的结果还提出了一种发现针对在其他感染性、炎症性和遗传性疾病进展中至关重要的RNA结构的药效基团的通用策略。
