A RATIONAL APPROACH TO ANTI-HIV DRUG DESIGN

我们的基本目标是改进设计抗艾滋病毒的方法 与细胞质HIV病毒有最大相互作用的药物 核酸（高活性）和与宿主的最小相互作用 细胞染色体DNA（低毒性）通过两种广泛的方法： 分子靶向-选择性结合艾滋病毒衍生的药物 核酸在细胞质中由于结构和动力学 病毒和宿主核酸之间的差异，以及 具有庞大基团的区室化药物， 强烈与病毒核酸和利用大小依赖性 分子进入细胞核的过程。 的基本思路 抗HIV药物的分子靶向如下：（i）通过使用范围 先进的技术，包括先进的核磁共振和分子 图形建模方法，现有化合物与 可以增强RNA的特异性二级结构特征， 新的化合物可以被设计出来， RNA：（ii）由于宿主细胞的DNA和RNA合成发生 在细胞核中，HIV病毒的复制代谢可以是 选择性地被破坏。 病毒RNA具有复杂的二级结构 （堆叠的单链、发夹双螺旋部分，和 具有不匹配或错配碱基的重叠区域）。 许多 具有不寻常的二级结构特征的区域代表 病毒RNA转化为病毒RNA的关键控制区域 前病毒DNA和非常有吸引力的设计目标， 抗艾滋病药物。 将对所有新的 合成化合物，比较它们对RNA的亲和力， 标准染色质制备。 更详细的动力学，NMR和 计算机模拟研究将在最活跃的 与特定二级结构紧密结合的化合物 RNA的特征，但与染色质的结合较弱。 基于这些 研究，新的化合物将设计与分子的帮助下， 建模方法 这些新化合物应该具有显著的 增强了对病毒RNA的亲和力。 药物的基本概念 细胞质中的区室化， 病毒核酸的过程可以被破坏， 分子从细胞质缓慢地进入细胞 核，许多有毒过程可以发生。 一系列分子 将被设计成具有大体积的取代基， 增强了与病毒RNA的结合 分配系数 所有新化合物将在合成中确定和调整 程序，使得潜在的药物实现良好的细胞质 浓度的 这些大分子进入细胞核 将通过荧光监测标准细胞系的 显微镜检查，以评估大小和运输之间的相关性 进入细胞核。

Our fundamental goal is to improve methods of designing anti-HIV drugs which have maximum interactions with cytoplasmic HIV viral nucleic acids (high activity) and minimum interactions with host cell chromosomal DNA (low toxicity) by two broad approaches: molecular targeting - drugs that selectively bind to HIV derived nucleic acids in the cell cytoplasm due to structural and dynamic differences between viral and host nucleic acids, and compartmentalization drugs with bulky groups that will interact strongly with viral nucleic acids and exploit the size dependence of partitioning of molecules into the nucleus. The basic ideas for molecular targeting of anti-HlV drugs follow: (i) by using a range of sophisticated techniques, including advanced NMR and molecular graphics-modeling methods, binding of existing compounds to specific secondary structural features of RNA can be enhanced and new compounds can be designed that specifically interact with viral RNA: (ii) since the DNA and RNA synthesis of the host cell occurs in the nucleus, the replicative metabolism of the HIV virus can be selectively disrupted. Viral RNA has a complex secondary structure (stacked single-strands, hairpin double-helical sections, and duplexed regions with unmatched or mismatched bases). Many of the regions with unusual secondary structural features represent critical control regions for the conversion of viral RNA into the proviral DNA and the very attractive targets for the design of anti-HIV drugs. Binding studies will be conducted for all new synthetic compounds to compare their affinity for RNA relative to a standard chromatin preparation. More detailed kinetics, NMR and computer modeling studies will be conducted on the most active compounds which bind strongly to specific secondary structural features of RNA but which bind weakly to chromatin. Based on these studies, new compounds will be designed with the aid of molecular modeling methods. These new compounds should have significantly enhanced affinity for the viral RNA. The basic idea for drug compartmentalization in the cytoplasm where important metabolic processes of viral nucleic acids can be disrupted is that bulky molecules pass quite slowly from the cell cytoplasm into the cell nucleus where many toxic processes can occur. A range of molecules will be designed with bulky substituents situated to maintain or actually enhance binding to viral RNA. Partition coefficients for all new compounds will be determine and adjusted in synthetic procedures such that the potential drugs achieve good cytoplasmic concentrations. Passage of these bulky molecules into the nucleus of a standard cell line will be monitored by fluorescence microscopy to evaluate the correlation between size and transport into the nucleus.