Small Molecule Inhibitors of HIV1 Nef Virulence Factor for Treatment of HIV_AIDS

 描述（由申请人提供）：虽然艾滋病毒/艾滋病可以用抗逆转录病毒药物治疗，但这些药物不能清除病毒，需要终身服用。最近，我们发现了一类全新的化合物，可以干扰HIV-1毒力因子Nef。这种病毒蛋白对HIV-1在体内复制、HIV感染细胞的免疫逃逸和AIDS进展至关重要。在过去一年，我们成功完成了STTR项目的第一阶段，该项目旨在开发适用于临床测试的Nef拮抗剂。我们与Fox Chase化学多样性中心合作，评估了我们最初的二苯基吡唑并二氮烯Nef抑制剂的50种类似物，其中一些显示出更紧密的Nef结合，同时保留了有效的抗逆转录病毒活性和改善的ADME特性。几种类似物阻止HIV感染的CD 4 + T细胞上的MHC-I的Nef介导的下调，导致自体抗HIV CD 8 + CTL的活化。这些数据表明，我们的Nef拮抗剂可以恢复患者自身免疫系统对HIV感染细胞的识别，作为功能性治愈的途径。在此II期申请中，我们将扩大Nef药物开发工作，具体目标如下：目标1：进行铅优化药物化学。基于在I期期间开发的SAR，我们建议合成100-150种新的Nef抑制剂类似物，以找到合适的化合物用于使用HIV感染的人源化小鼠在Aim 3中进行体内测试。我们的方法将采用基于结构的设计，同时考虑模拟ADMET和PK属性并行。还将测试所有类似物的Nef结合亲和力、对Nef介导的HIV复制增强的作用以及Nef在HIV感染的患者细胞中对CD 4和MHC-I下调的逆转。目的2：通过体外和体内ADMET评估确保合适的药物性质。每年将使用体外ADMET试验（包括微粒体稳定性、α 3A 4抑制、溶解度和血浆蛋白结合）评价多达15种符合Nef结合和功能标准（目标1）的化合物。将在小鼠PK研究中评价3 - 6种（IV）和2-3种（PO）化合物，其中2-3种最有希望的化合物将进行体外非GLP安全性试验。这些数据对于选择用于在HIV/AIDS的人源化小鼠模型中进行体内测试的最佳化合物是必不可少的（目的3）。目的3：检验Nef拮抗剂可以在AIDS的人源化小鼠模型中抑制HIV复制和T细胞损失的假设。与感染野生型病毒的小鼠相比，感染Nef缺失的HIV-1的人源化小鼠表现出显著更低的病毒载量和显著更少的T细胞消耗。因此，在前两个目标中确定的最有希望的化合物将被给予人源化小鼠，以监测对HIV复制，CD 4 + T细胞损失和免疫系统功能的影响。这些目标的成功完成将提供一个全面的包，以支持先进的开发，在大型动物中的安全性测试，并推动该项目进一步向IND提交，在正常志愿者中进行I期安全性测试。

 DESCRIPTION (provided by applicant): While HIV/AIDS can be managed with antiretroviral drugs, these agents do not clear the virus and require life-long administration. Recently, we discovered a completely new class of compounds that interfere with the HIV-1 virulence factor called Nef. This viral protein is critical to HIV-1 replication in vivo, immune escape of HIV-infected cells, and AIDS progression. During the past year, we successfully completed Phase I of our STTR project aimed at development of Nef antagonists suitable for clinical testing. Working with the Fox Chase Chemical Diversity Center, we evaluated 50 analogs of our original diphenylpyrazolodiazene Nef inhibitor, some of which display tighter Nef binding while retaining potent antiretroviral activity and improved ADME properties. Several analogs prevent Nef-mediated downregulation of MHC-I on HIV-infected CD4+ T-cells, resulting in activation of autologous anti-HIV CD8+ CTLs. These data suggest that our Nef antagonists may restore recognition of HIV-infected cells by the patient's own immune system as a path to functional cure. In this Phase II application, we will expand our Nef drug development efforts with the following Specific Aims: Aim 1: Perform lead optimization medicinal chemistry. Based on SAR developed during Phase I, we propose to synthesize 100-150 new Nef inhibitor analogs to find suitable compounds for in vivo testing in Aim 3 using HIV-infected humanized mice. Our approach will employ structure-based design while considering analog ADMET and PK properties in parallel. All analogs will also be tested for Nef binding affinity, effects on Nef-mediated enhancement of HIV replication and reversal of CD4 and MHC-I downregulation by Nef in HIV-infected patient cells. Aim 2: Ensure suitable drug properties via in vitro and in vivo ADMET evaluation. Up to 15 compounds per year that meet Nef-binding and functional criteria (Aim 1) will be evaluated using in vitro ADMET assays including microsomal stability, CYP 3A4 inhibition, solubility and plasma protein binding. Three to six (IV) and 2-3 (PO) compounds will be evaluated in mouse PK studies, with the 2-3 most promising compounds advancing to in vitro non-GLP safety assays. These data are essential for choosing the best compounds for in vivo testing in humanized mouse models of HIV/AIDS (Aim 3). Aim 3: Test the hypothesis that Nef antagonists can suppress HIV replication and T-cell loss in a humanized mouse model of AIDS. Humanized mice infected with Nef-deleted HIV-1 exhibit dramatically lower viral loads and substantially less T cell depletion than those infected with wild-type virus. The most promising compounds identified in the first two Aims will therefore be administered to humanized mice to monitor effects on HIV replication, CD4+ T cell loss and immune system function. Successful completion of these goals will provide a comprehensive package to support advanced development, safety testing in large animals and advance the project further toward an IND submission for Phase I safety testing in normal volunteers.