Novel anti-HIV compounds targeting the HIV-1 matrix protein

摘要 因为耐药菌株的出现和与当前 在治疗方面，对艾滋病毒-1复制的新抑制剂的需求依然存在。HIV-1基质蛋白(MA)是一种 病毒的基本成分，在病毒的组装中具有既定的作用。结合使用虚拟 筛选，表面等离子共振分析，和抗病毒测试，我们第一个发现了小分子药物- 比如与HIV-1 MA蛋白结合并具有广泛抗HIV特性的分子。组合 计算机辅助药物设计技术和药物化学，我们设计和合成了下一步 产生与HIV-1 MA相互作用并抑制病毒的化合物。我们目前的先导化合物，MTI-14 与HIV-1MA结合(SPR和核磁共振判断)，毒性低，具有中微摩尔效力。此外， MA突变病毒对MTI-14的敏感性降低，表明 化合物是通过破坏MA的功能来调节的。令人兴奋的是，初步的行动机制 研究还表明，MTI-14通过阻止HIV-1Env并入新的病毒颗粒而发挥作用。我们 建议进一步优化这种MA靶向抑制化合物，以达到临床相关的效力，同时 使用这些抑制剂作为分子工具来剖析HIV-1复制周期中MA蛋白的生物学。 拟议研究的目标将通过三个独立但高度综合的具体目标来实现。在……里面 目标1，通过结构指导设计和药物化学方法优化引线，其 将测量动力学属性，包括开启和关闭速率和结合亲和力(Kd)，以及它们的结合位置 用化学位移核磁共振和竞争荧光偏振分析进行了验证。此外，新的化学类型 将使用基于配体的筛选方法寻找。在目标2中，抗病毒的效力和广度 将对线索进行评估。对其毒性的评估将同时进行。在目标3中，我们将表演 详细的作用机制研究，产生抗药性突变体，并确定新的结构 用结晶学和核磁共振方法研究了与MA形成络合物的缓蚀剂。数据来自 这些研究将被用于设计具有高遗传障碍的有效的下一代MA靶向抑制剂 和独特的行动模式。这些化合物还将作为HIV-1MA的新型分子探针 功能。

ABSTRACT Because of the emergence of drug-resistant strains and the cumulative toxicities associated with current therapies, demand remains for new inhibitors of HIV-1 replication. The HIV-1 matrix protein (MA) is an essential viral component with established roles in the assembly of the virus. Using a combination of virtual screening, surface plasmon resonance analysis, and antiviral testing, we were the first to identify small drug- like molecules that bind to the HIV-1 MA protein and that possess broad-range anti-HIV properties. Combining computer-aided drug design techniques and medicinal chemistry, we have designed and synthesized next generation compounds that interact with HIV-1 MA and inhibit the virus. Our current lead compound, MTI-14 binds to HIV-1 MA (as judged by SPR and NMR), has low toxicity, and has mid-micromolar potency. Moreover, MA mutant viruses display reduced sensitivity to MTI-14, demonstrating that the antiviral action of the compound is mediated through disruption of the functions of MA. Excitingly, preliminary mechanism-of-action studies also suggest that MTI-14 functions by blocking HIV-1 Env incorporation into new viral particles. We propose to further optimize this MA-targeted inhibitory compound to potencies that are clinically relevant, while using these inhibitors as molecular tools to dissect the biology of the MA protein in the HIV-1 replication cycle. The goals of the proposed studies will be accomplished by three discrete but highly integrated specific aims. In Aim 1, the leads will be optimized through structure-guided design and medicinal chemistry approaches, their kinetic properties including on- and off-rates and binding affinities (KD) will be measured, and their binding site verified by chemical shift NMR and competitive fluorescent polarization assays. In addition, novel chemotypes will be sought using ligand-based screening approaches. In Aim 2, the antiviral potency and breadth of the leads will be assessed. Evaluation of their toxicity will be performed in parallel. In Aim 3, we will perform detailed mechanism-of-action studies, generate resistance mutants, and determine the structures of the new inhibitors in complex with MA using crystallographic and nuclear magnetic resonance methods. The data from these studies will be used in designing potent next-generation MA-targeted inhibitors with high genetic barriers and unique modes of action. Such compounds will also serve as novel molecular probes for HIV-1 MA functions.