HIV Latency Reversal Through Novel, Potent PKC Modulators

摘要 虽然抗逆转录病毒疗法（ART）减缓了疾病进展，但ART是一种终身疗法，需要严格的治疗。 避免病毒耐药性的医疗依从性，并且与显著的成本和长期的医疗费用相关。 不良影响此外，ART不是治疗性的，因为它没有消除复制能力的储存库。 病毒因此，如果停止抗逆转录病毒疗法，艾滋病毒可能从宿主中出现并迅速传播，导致疾病 向艾滋病发展。清除这些潜伏感染细胞库的一种潜在策略是使用 一种踢和杀的方法，在这种方法中，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感染细胞的额外疗法的“杀伤”。 潜伏期逆转剂（LRA）已被用于“踢”或诱导潜伏细胞的HIV表达，但迄今为止， 受到次优功效、药物毒性和/或生物分布问题的限制。在这次更新申请中， 我们使用新的LRA概念来解决这些问题，以提高疗效和耐受性， 生物分布我们研究蛋白激酶C（PKC）调节剂，这是最有力和有效的 LRA，特别关注表现最好的-苔藓抑素-1，prostratin和巨大戟二萜醇酯， 新的LRA的ingenane和tigliane家庭。我们已经发现苔藓抑素-1化学转化为 前药（缓释）形式产生具有上级活性和改善的耐受性的新LRA。因此 这一更新申请的目标是推进这一新概念和新类别的化学研究。 合成前体药物LRA使用高度协作的团队，具有新颖的基于计算机的设计的专业知识， 合成、药物化学、最先进的体外测定和复杂的体内动物建模。 在人源化小鼠中，最佳前药LRA将与“杀死”方法（自然杀伤细胞）结合使用 潜在感染艾滋病毒的人，以评估踢和杀方法的有效性。我们会实现我们的目标 1）在体外和体内评价基于以下的潜伏期逆转的新概念： 新的LRA前体药物，2）确定导致HIV潜伏的特定PKC亚型和非PKC途径 逆转，和3）研究新的和已知的LRA的协同作用，提高艾滋病毒库消耗。集体 这些研究将推动我们独特的、上级临床前LRA进入临床试验， 为执行根除艾滋病毒的“踢和杀”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.