Intervening with Latent HIV-1 Infection using Gnidimacrin

虽然抗逆转录病毒疗法(ART)已经成功地控制了HIV-1阳性患者的病毒复制 就个人而言，艾滋病毒-1是被抑制而不是根除的。据认为，潜伏的HIV-1宿主是 对持续感染负有责任。因此，从潜伏的宿主中消除HIV-1的策略是 需要治愈/从功能上治愈持续的HIV-1感染。我们的长期目标是发展一种有选择性的 蛋白激酶C激动剂格列地马林(GM)作为根除HIV-1的辅助治疗。通用汽车是一家 异常强大的天然产品，在皮摩尔浓度下激活潜伏的HIV-1复制。它不是 对未受感染的细胞有毒，直到达到微摩尔浓度。通用汽车的威力至少比 研究最多的是组蛋白去乙酰酶抑制剂伏立诺斯特(SAHA)，用于潜伏的HIV-1激活。更重要的是， 在潜伏感染的细胞系模型中，GM诱导的病毒产量大约是SAHA的10倍。 尽管SAHA被证明可以扰乱HIV-1潜伏期，但它似乎在减少潜伏期HIV方面无效 病毒潜伏激活后，病毒通过自体CTL反应或细胞病变效应感染宿主。因此，它是 提出了其他策略，如激活CTL，可能需要消除潜伏感染的细胞 在萨哈治疗之后。由于GM在潜伏的病毒激活方面比SAHA强得多，我们假设 GM的强烈激活将导致HIV-1潜伏感染细胞产生强劲的HIV-1，这将 使潜伏感染的细胞对激活的潜伏细胞的CTL反应和/或细胞病变效应敏感 病毒。因此，本研究的目的是确定GM在减少潜伏病毒库方面的作用 使用HIV-1阳性患者的原代PBMC，并建立关键的药理学和毒理学 未来体内临床前研究所需的资料。这些目标将通过以下方式实现 具体目标：1)确定GM在减少初级静息CD_4中HIV-1蓄积方面的有效性 来自病人的细胞。我们实现这一目标的方法是确定潜伏期 感染的CD4细胞对GM激活的潜伏病毒的CTL反应和细胞病变效应；2)建立 转基因药物临床前药代动力学和毒理学研究概况。由于PKC属于丝氨酸/苏氨酸家族 参与重要的细胞功能，这是至关重要的确定临床前的药理学和 转基因动物体内药效研究前的毒理学特性。小动物模型将被用于 建立转基因药物的药代动力学和毒理学模型。除了其强大的抗潜伏期活性外，GM 与HDAC抑制剂相比，它还具有优势，因为它可以在PM抑制HIV-1R5病毒的再次感染 浓度。拟议的研究将使我们能够确定转基因是否可以潜伏地减少/消除 在没有额外免疫干预的情况下感染静息的CD4细胞。因此通用汽车有可能成为一家 用于根除HIV-1的有吸引力的候选药物。

While anti-retroviral therapy (ART) has been successful in controlling virus replication in HIV-1 positive individuals, HIV-1 is suppressed rather than eradicated. It is believed that latent HIV-1 reservoirs are responsible for the persistent infection. Therefore, strategies that eliminate HIV-1 from latent reservoirs are needed to cure/functionally cure persistent HIV-1 infection. Our long term goal is to develop a selective protein kinase C agonist gnidimacrin (GM) as an adjuvant therapeutic for HIV-1 eradication. GM is an unusually potent natural product that activates latent HIV-1 replication at picomolar concentrations. It is not toxic to uninfected cells until it reaches micromolar concentrations. GM is at least 4 log10 more potent than the most well studied histone deacetylase inhibitor vorinostat (SAHA) for latent HIV-1 activation. More importantly, GM induces approximately 10-fold more virus production than SAHA in a latently infected cell line model. Although SAHA was shown to disrupt HIV-1 latency, it appears to be ineffective in reducing latent HIV reservoirs through autologous CTL responses or cytopathic effect following latent viral activation. Thus, it was proposed that other strategies, such as CTL activation, might be needed to eliminate latently infected cells following SAHA treatment. Since GM is much stronger than SAHA in latent virus activation, we hypothesize that strong activation by GM will result in robust HIV-1 production from HIV-1 latently infected cells, which will render the latently infected cells susceptible to CTL responses and/or cytopathic effects of the activated latent viruses. Therefore, the goal of this study is to determine the effect of GM on reducing latent viral reservoirs using primary PBMCs from HIV-1 positive patients, and to establish crucial pharmacology and toxicology profiles that are needed for future in vivo pre-clinical studies. The goals will be achieved with the following Specific Aims: 1) determining the effectiveness of GM on reducing HIV-1 reservoirs in primary resting CD4 cells from patients. Our approaches to accomplish this aim are to determine the susceptibility of latently infected CD4 cells to CTL responses and cytopathic effects of GM-activated latent viruses; 2) establishing the preclinical pharmacokinetic and toxicology profiles of GM. Since PKC belongs to a family of serine/threonine kinases involved in vital cellular functions, it is essential to determine the preclinical pharmacological and toxicological properties of GM before in vivo animal studies for efficacy. Small animal models will be used to establish the pharmacokinetic and toxicology profiles of GM. In addition to its strong anti-latency activity, GM also has advantage over HDAC inhibitors in that it inhibits re-infection of HIV-1 R5 viruses at pM concentrations. The proposed study will allow us to determine whether GM can reduce/eliminate latently infected resting CD4 cells without additional immunological interventions. Thus GM has the potential to be an attractive drug candidate for HIV-1 eradication.