Design, optimization and mechanism of action of novel HIV-1 entry inhibitors

 描述(由申请人提供)：随着耐药菌株的出现和与当前疗法相关的累积毒性，对艾滋病毒-1复制的新抑制剂的需求仍然存在。抑制艾滋病毒-1进入是一种有吸引力的治疗方法，对艾滋病毒/艾滋病治疗方案具有广泛的影响。结合计算机辅助药物设计技术，基于化合物的新的力场表示，与药物化学相结合，我们设计并合成了新型的HIV-1进入抑制剂，它们代表了HIV-1进入抑制剂类别的5个新的不同的核心化学类型。这些化合物毒性低，纳摩尔效力(0.6-100 nm)，并且对HIV-1具有特异性。此外，我们已经确定了一种化合物，与目前正在开发的同类最佳进入抑制剂相比，其代谢稳定性有所改善。在这项研究中，我们建议进一步开发这些新的进入抑制剂化合物以达到临床相关的效力，并改善它们的ADME/PK谱，同时也将它们作为分子工具来剖析HIV-1进入宿主细胞的生物学。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。在目标1中，将通过电子计算方法优化先导化合物，包括生物等位元替代和类药物质量预测，并与药物化学相结合。在目标2中，将评估类似物的抗病毒效力、广度和特异性。对其毒性的评估将同时进行。ADME和PK将通过体外和体内方法进行评估，其有效性将在人源化的骨-肝-胸腺(BLT)小鼠HIV-1感染模型中确定。在目标3中，我们将进行详细的作用机制研究，产生抗性突变体，并使用互补的结构技术研究化合物的结合位置，包括X射线结晶学、低温电子显微镜和负染色单粒子电子显微镜。这些研究的数据将用于未来开发具有高遗传障碍和独特作用模式的下一代Env靶向抑制剂。这些化合物还将作为HIV-1 Env功能的分子探针，除了揭示难以捉摸的脆弱部位的细节外，还提供了对Env分子机器进行的一系列复杂结构转变的新见解。

 DESCRIPTION (provided by applicant): With the emergence of drug-resistant strains and the cumulative toxicities associated with current therapies, demand remains for new inhibitors of HIV-1 replication. The inhibition of HIV-1 entry is an attractive therapeutic approach with broad implications for HIV/AIDS treatment regimens. Combining computer-aided drug design techniques, based upon novel force field representations of compounds, with medicinal chemistry, we have designed and synthesized novel inhibitors of HIV-1 entry, which represent 5 new distinct core chemotypes for the HIV-1 entry inhibitor class. These compounds have low toxicity, nanomolar potency (0.6-100 nM), and are specific to HIV-1. Moreover, we have identified a compound with improved metabolic stability over the current best-in- class entry inhibitor in development. In this study, we propose to further develop these novel entry inhibitor compounds to potencies that are clinically relevant and improve their ADME/PK profiles, whilst also using them as molecular tools to dissect the biology of HIV-1 entry into host cells. 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 in silico approaches, including bioisosteric replacement and prediction of drug-like qualities, coupled to medicinal chemistry. In Aim 2, the antiviral potency, breadth, and specificity of the analogues will be assessed. Evaluation of their toxicity will be performed in parallel. ADME and PK will be assessed via in vitro and in vivo methodologies, and the efficacy will be determined in the humanized bone-liver-thymus (BLT) mouse model of HIV-1 infection. In Aim 3, we will perform detailed mechanism-of-action studies, generate resistance mutants, and investigate the binding site of the compounds using complementary structural techniques including X-ray crystallography, cryo-EM, and negative-stain single-particle electron microscopy. The data from these studies will be used in the future development of next-generation Env-targeted inhibitors with high genetic barriers and unique modes of action. Such compounds will also serve as molecular probes for HIV-1 Env functions, providing novel insight into the complex series of structural transitions through which the Env molecular machine proceeds, in addition to illuminating the details of an elusive site of vulnerability.