Towards HIV eradication: New concepts and potent compounds for PKC-mediated latency reversal

摘要 虽然抗逆转录病毒疗法(ART)延缓了疾病的发展，但ART是一种终生疗法。值得注意的是，艺术 是无法治愈的。它不会消除复制能力强的病毒的蓄水池。因此，当艺术停止时， 艾滋病毒从水库中出现并迅速传播，导致疾病向艾滋病发展。一种策略 要清除这些潜伏感染细胞的蓄水池，持续感染的主要来源(如果不是唯一的来源)是 使用一种先踢后杀的方法，在这种方法中，潜伏细胞被“踢”或从潜伏期激活，从而允许它们随后的 通过病毒的细胞病变效应、免疫效应细胞或针对HIV的额外疗法来“杀死”或清除- 被感染的细胞。为此，我们展示了概念验证，即具有唯一有效延迟的踢杀策略 在人源化的小鼠模型中，逆转药物(LRA)和NK细胞显著靶向HIV储存库，导致 成为治疗中断的里程碑。我们的下一个目标是进一步增强我们的踢杀组件，以 根除抗逆转录病毒治疗期间存在的所有具有复制能力的病毒库。我们将设计、合成和 研究有希望的新化合物，这是迄今为止发现的最好的踢脚成分，总体目标是 产生卓越的LRA和协同策略，高效、安全地清除体内潜伏的艾滋病毒。LRAS 已经被用来从潜伏细胞中诱导HIV表达，但到目前为止一直受到效果不佳的限制， 耐受性问题，和/或生物分布问题。在这里，我们使用新的上帝抵抗军概念来解决这些问题 以及改善疗效、耐受性和控制生物分布的结构。我们研究了蛋白激酶C(PKC) 调制器，这是迄今为止报道的最有效和最有效的LRA。在这个家庭里，我们有 确定了迄今报道的最好的表现，灵感来自bryostatin-1，prostratin和ingenol esters以及新的LRA 异丁烷、三丁烷和环氧三丁烷家族的支架。我们发现，碳的化学转化 将Bryostatin-1转化为前药(缓释)版本导致了具有更高活性(60%比98%)的新型LRA和 显著提高耐受性(治疗窗口增加20倍)。因此，我们的目标是推进这项研究 这些新的LRA和化学合成的前药使用一个高度合作的团队，在新的专业知识 基于计算机的设计、合成、药物化学、最先进的体外试验和复杂的体内试验 动物模型。为了将这一计划推向临床，最佳的LRA和前药将用于 结合“杀死”方法(自然杀伤细胞)对潜伏感染艾滋病毒的人源化小鼠进行评估 踢杀策略的有效性。我们将通过以下具体目标实现目标：1) 基于新型LRA支架的新一代潜伏期反转剂的体内外评价 前药，代表迄今报道的最有效和耐受性最好的LRA，2)定义和选择性 激活PKC亚型以增强HIV潜伏期逆转和提高耐受性，以及3)开发新的 采取协同战略，消灭艾滋病毒携带者。总的来说，这些研究将推动我们独特的和 卓越的临床前LRA用于临床测试。

ABSTRACT Although anti-retroviral therapy (ART) slows disease progression, ART is a life-long therapy. Significantly, ART is not curative. It does not eliminate reservoirs of replication-competent virus. Thus, when ART is discontinued, HIV emerges from reservoirs and rapidly spreads, leading to disease progression towards AIDS. One strategy for clearing these reservoirs of latently infected cells, the principal if not sole source of continued infection, is to use a kick and kill approach, in which latent cells are “kicked” or activated from latency, allowing their subsequent “killing” or clearance by viral cytopathic effects, immune effector cells or additional therapies targeted at HIV- infected cells. To this end, we show proof-of-concept that a kick and kill strategy with uniquely effective latency reversing agents (LRA) and NK cells remarkably targets the HIV reservoir in a humanized mouse model, leading to a milestone of treatment interruption. Our next goal is to further enhance our kick and kill components to eradicate all replication-competent reservoirs of virus present during ART. We will design, synthesize, and investigate promising new compounds, the best kick components uncovered thus far, with the overall goal of producing superior LRAs and synergistic strategies that efficiently and safely purge the latent HIV in vivo. LRAs have been used to induce HIV expression from latent cells, but thus far have been limited by suboptimal efficacy, tolerability issues, and/or biodistribution concerns. Here, we address these problems using new LRA concepts and structures to improve efficacy, tolerability and control biodistribution. We investigate protein kinase C (PKC) modulators, which are the most potent and efficacious LRAs reported thus far. Within this family we have identified the best performers yet reported, inspired by bryostatin-1, prostratin and ingenol esters and new LRA scaffolds of the ingenane, tigliane, and epoxy-tigliane families. We have found that the chemical conversion of bryostatin-1 into a prodrug (slow-release) version results in a novel LRA with superior activity (60% v. 98%) and significantly improved tolerability (20-fold increase in therapeutic window). Thus, our goal is to advance this study of these new LRAs and chemically synthesized prodrugs using a highly collaborative team with expertise in novel computer-based design, synthesis, medicinal chemistry, state-of-the-art in vitro assays, and sophisticated in vivo animal modeling. To move this program toward clinical entry, optimal LRAs and prodrugs will be used in conjunction with a “kill” approach (natural killer cells) in humanized mice latently infected with HIV to assess the efficacy of the kick and kill strategy. We will accomplish our goals through the following Specific Aims: 1) Evaluate in vitro and in vivo new generation latency reversal agents based on new LRA scaffolds and their prodrugs, representing the most effective and best tolerated LRAs reported thus far, 2) Define and selectively activate PKC isoforms that enhance HIV latency reversal and improve tolerability, and 3) Develop new synergistic strategies to deplete the HIV reservoir. Collectively these studies will advance our unique and superior preclinical LRAs towards clinical testing.