Intervening with Latent HIV-1 Infection using Gnidimacrin

产品说明：虽然抗逆转录病毒疗法（ART）已成功地控制HIV-1阳性个体的病毒复制，但HIV-1是被抑制而不是根除的。据认为，潜伏的HIV-1储库是造成持续感染的原因。因此，需要从潜伏储库中消除HIV-1的策略来治愈/功能性治愈持续性HIV-1感染。我们的长期目标是开发一种选择性蛋白激酶C激动剂gnidimacrin（GM）作为HIV-1根除的辅助治疗药物。GM是一种非常有效的天然产物，在皮摩尔浓度下可激活潜伏的HIV-1复制。在达到微摩尔浓度之前，它对未感染的细胞没有毒性。对于潜伏的HIV-1激活，GM比研究最充分的组蛋白去乙酰化酶抑制剂伏立诺他（SAHA）至少强4 log 10。更重要的是，在潜伏感染的细胞系模型中，GM诱导的病毒产量比SAHA多约10倍。尽管SAHA显示破坏HIV-1潜伏期，但其似乎在通过自体CTL应答或潜伏病毒活化后的细胞病变效应减少潜伏HIV储库方面无效。因此，有人提出，可能需要其他策略，如CTL激活，以消除潜伏感染的细胞后SAHA治疗。由于GM在潜伏病毒活化方面比SAHA强得多，我们假设GM的强活化将导致HIV-1潜伏感染细胞产生稳健的HIV-1，这将使潜伏感染细胞对活化的潜伏病毒的CTL应答和/或细胞病变效应敏感。因此，本研究的目的是使用来自HIV-1阳性患者的原代PBMC确定GM对减少潜伏病毒储库的影响，并建立未来体内临床前研究所需的关键药理学和毒理学特征。这些目标将通过以下具体目标实现：1）确定GM在减少患者原代静息CD 4细胞中HIV-1储库方面的有效性。我们的方法来实现这一目标是确定潜在感染的CD 4细胞的敏感性CTL反应和GM激活的潜伏病毒的细胞病变效应; 2）建立GM的临床前药代动力学和毒理学特征。由于PKC属于参与重要细胞功能的丝氨酸/苏氨酸激酶家族，因此在体内动物研究疗效之前确定GM的临床前药理学和毒理学特性至关重要。将使用小动物模型来建立GM的药代动力学和毒理学特征。除了其强的抗潜伏活性之外，GM还具有优于HDAC抑制剂的优势，因为它在pM浓度下抑制HIV-1 R5病毒的再感染。拟议的研究将使我们能够确定GM是否可以减少/消除潜伏感染的静息CD 4细胞，而无需额外的免疫干预。因此，GM有可能成为一种有吸引力的HIV-1根除候选药物。

DESCRIPTION: 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 toxi 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 effet 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.