HIV Latency Reversal Through Novel, Potent PKC Modulators

摘要 虽然抗逆转录病毒疗法(Art)延缓了疾病的进展，但它是一种终生疗法，它需要严格的 医疗合规以避免病毒耐药性，并与巨大的成本和长期的医疗相关 不良影响。此外，抗逆转录病毒疗法不能治愈，因为它不能消除具有复制能力的储存库。 病毒。因此，如果停止抗逆转录病毒治疗，艾滋病毒可能会从水库中涌出并迅速传播，从而导致疾病 爱滋病的进展。清除这些潜伏感染细胞的一个潜在策略是使用 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艾滋病毒感染细胞的额外治疗来“杀死”。 潜伏期反转剂(LRA)已被用来“踢”或诱导潜伏细胞中的HIV表达，但到目前为止 受到次优疗效、药物毒性和/或生物分布问题的限制。在此续签申请中， 我们使用一种新的上帝抵抗军概念来解决这些问题，以提高疗效、耐受性和控制力 生物分布。我们研究了蛋白激酶C(PKC)调节剂，它是最有效和最有效的 LRA，特别关注表现最好的-bryostatin-1，prostratin和ingenol酯，并可能 异丙烷和三丁烷家族的新LRA。我们发现，Bryostatin-1的化学转化为 前药(缓释)版本导致了具有更高活性和更好耐受性的新型LRA。因此， 本次更新申请的目的是为了促进这一新概念和新类别的化学研究 使用一个高度协作的团队合成前药LRA，该团队在基于计算机的新型设计方面具有专业知识， 合成、药物化学、最先进的体外分析和复杂的活体动物模型。 最佳前体药物LRAs将与人源化小鼠的“杀伤”方法(自然杀伤细胞)结合使用 潜伏感染艾滋病毒，以评估踢杀方法的效果。我们将实现我们的目标 通过以下具体目标：1)在体外和体内评估一种新的基于以下因素的潜伏期逆转概念 新的LRA前体药物，2)定义导致HIV潜伏期的特定PKC亚型和非PKC途径 逆转，以及3)研究新的和已知的LRA的协同作用，从而增强艾滋病毒储备库的枯竭。集体地 这些研究将推动我们独特和卓越的临床前LRA进行临床测试，并提供 为执行消灭艾滋病毒的踢杀方法所急需的信息。

ABSTRACT Although anti-retroviral therapy (ART) slows disease progression, ART is a life-long therapy, which requires strict medical compliance to avoid viral resistance and is associated with significant costs and long-term medically adverse effects. In addition, ART is not curative because it does not eliminate reservoirs of replication-competent virus. Thus, if ART is discontinued, HIV can emerge from reservoirs and rapidly spread, leading to disease progression towards AIDS. One potential strategy for clearing these reservoirs of latently infected cells is to use a kick and kill approach, in which latent cells are “kicked” or activated from latency, allowing their subsequent “killing” by viral cytopathic effects, immune effector cells or additional therapies targeted at HIV-infected cells. Latency reversing agents (LRA) have been used to “kick” or induce HIV expression from latent cells, but thus far have been limited by suboptimal efficacy, drug toxicity, and/or biodistribution issues. In this renewal application, we address these problems using a new LRA concept to improve efficacy and tolerability and control biodistribution. We investigate protein kinase C (PKC) modulators, which are the most potent and efficacious LRAs, focusing particularly on the best-performers - bryostatin-1, prostratin and ingenol esters and potentially new LRAs of the ingenane and 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 and improved tolerability. Thus, the goal of this renewal application is to advance the study of this new concept and new class of chemically synthesized prodrug LRAs 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. Optimal prodrug LRAs 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 approach. We will accomplish our goals through the following Specific Aims: 1) Evaluate in vitro and in vivo a new concept for latency reversal based on new LRA prodrugs, 2) Define the specific PKC isoforms and non-PKC pathways that contribute to HIV latency reversal, and 3) Investigate synergies of new and known LRAs that enhance HIV reservoir depletion. Collectively these studies will advance our unique and superior preclinical LRAs towards clinical testing and provide information critically needed for implementation of kick and kill approaches for HIV eradication.