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）可以减缓疾病进展，但ART是一种终身治疗。重要的是，艺术 是没有疗效的它并不能消除具有复制能力的病毒的储存库。因此，当抗逆转录病毒疗法停止时， 艾滋病毒从宿主中出现并迅速传播，导致疾病进展为艾滋病。一种策略 为了清除这些潜伏感染细胞的储库，如果不是唯一的持续感染源，主要是 使用踢和杀的方法，其中潜伏细胞被“踢”或从潜伏期激活， 通过病毒致细胞病变效应、免疫效应细胞或靶向HIV的其他疗法的“杀伤”或清除， 被感染的细胞为此，我们展示了一个概念验证，即具有独特有效延迟的踢杀策略 逆转剂（LRA）和NK细胞在人源化小鼠模型中显著靶向HIV储库， 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， 根除ART期间存在的所有具有复制能力的病毒库。我们将设计，合成， 研究有前途的新化合物，迄今为止发现的最好的踢成分，总体目标是 产生有效和安全地清除体内潜伏的HIV的上级LRA和协同策略。LRAs 已被用于诱导潜伏细胞的HIV表达，但迄今为止受到次优功效的限制， 耐受性问题和/或生物分布问题。在这里，我们使用新的LRA概念来解决这些问题 和结构以改善功效、耐受性和控制生物分布。我们研究蛋白激酶C（PKC） 调节剂，这是迄今为止报道的最有效和最有效的LRA。在这个家庭里， 确定了迄今为止报道的最佳表现，灵感来自苔藓抑素-1，prostratin和巨大戟二萜醇酯和新的LRA 巨大戟烷、惕各烷和环氧-惕各烷家族的骨架。我们发现， 将苔藓抑素-1转化为前药（缓释）形式产生了具有上级活性的新型LRA（60%对98%）， 显著改善耐受性（治疗窗增加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协作的团队， 基于计算机的设计，合成，药物化学，最先进的体外测定，以及复杂的体内 动物模型为了将该项目推向临床，将使用最佳的LRA和前药， 在潜伏感染HIV的人源化小鼠中结合“杀死”方法（自然杀伤细胞）来评估 kick and kill strategy策略的有效性。我们将通过以下具体目标来实现我们的目标：1） 在体外和体内评估基于新的LRA支架的新一代潜伏期逆转剂及其 前药，代表迄今为止报道的最有效和最耐受的LRA，2）定义和选择性地 激活PKC亚型，增强HIV潜伏期逆转并提高耐受性，以及3）开发新的 协同战略，以消除艾滋病毒库。总的来说，这些研究将推动我们独特的， 临床试验的上级临床前LRA。

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.