Novel Kinase and Nanoformulated Protease Inhibitors for Eradication of CNS HIV-1

描述（由申请人提供）：本申请基于URMC-099可显著增强长效抗逆转录病毒疗法的抗病毒活性的偶然发现。URMC-099被开发为正在开发的用于治疗HIV-1相关神经认知障碍（HAND）的新型领先的一流混合谱系激酶（MLK）抑制剂，意外地发现URMC-099可增强纳米利托那韦增强的阿扎那韦（nanoATV/r）的抗逆转录病毒作用。这种药物组合导致残留HIV-1感染的显著减少。URMC-099通过影响调节内体囊泡运输的Rab家族蛋白的表达，增强病毒生命周期期间nanoATV/r与病毒颗粒之间的相互作用，促进nanoATV/r治疗效果。在此，我们将确定来自URMC-099的MLK抑制是否参与ATV/r在内体中的作用，或者该发现是否是由于对该试剂靶向的其他激酶的影响。我们最近证明URMC-099在体内减少神经炎症并对HIV-1蛋白具有神经保护作用，并且非选择性MLK抑制剂CEP-1347可以增加HIV-1感染患者中的ATV积累和半衰期，这预示着这种化疗组合可以安全地治疗HIV-1感染患者。 实现CNS中的病毒根除，同时保护神经稳态。我们提出了三个具体的目标，以进一步描绘这些相互作用：（1）我们将阐明的途径负责URMC-099促进单核吞噬细胞中纳米配制抗逆转录病毒应答的（机制）（MP;单核细胞衍生的巨噬细胞[MDM]和小胶质细胞）与MLK和URMC-099靶向的其他相关激酶的关系（包括介导炎症的激酶），并决定是否可以根除或仅仅抑制病毒。(2)在我们的HAND模型中评估目标1中确定的Rab家族成员对URMC-099和nanoART的潜在药物毒性的影响。为此，我们将研究URMC-099和nanoART治疗后，其在调节暴露于HIV-1神经毒素的MP和突触网络中的作用。我们将使用体内模型来进一步了解Rabs的作用，因为它们与MLK激活和突触树突损伤有关，目的是保护大脑免受进一步损伤。最后，在（3）中，我们将验证纳米配制的抗逆转录病毒疗法（nanoART）和URMC-099在我们的人源化CD 34移植的neuroAIDS模型中清除CNS中持续感染的病毒储库的效用。这些目标将共同推进我们根除中枢神经系统MP长期持续感染的方法，同时保留疾病中的突触结构和功能。

DESCRIPTION (provided by applicant): This application is based on the serendipitous finding that URMC-099 can significantly boost antiviral activities of long acting antiretroviral therapy. Developed as a novel lead first-in-class mixed lineage kinase (MLK) inhibitor under development for use against HIV-1 associated neurocognitive disorders (HAND), URMC-099 was unexpectedly found to potentiate antiretroviral actions of nanoformulated ritonavir- boosted atazanavir (nanoATV/r). This drug combination led to a marked reduction of residual HIV-1 infection. URMC-099 facilitated nanoATV/r therapeutic effects by affecting the expression of the Rab family proteins that regulate endosomal vesicle trafficking, augmenting interactions between nanoATV/r and viral particles during the viral life cycle. Herein we will determine whether MLK inhibition from URMC-099 is involved in ATV/r's actions in the endosome, or if the findings are due to effects on other kinases targeted by this agent. Our recent demonstration that URMC-099 reduces neuroinflammation and is neuroprotective against HIV-1 proteins in vivo, and that the non-selective MLK inhibitor, CEP-1347 can increase ATV accumulation and half-life in HIV-1 infected patients bodes very well for this combination of chemotherapy to safely achieve viral eradication in the CNS while protecting neural homeostasis. We propose three specific aims to further delineate these interactions: (1) We will elucidate the pathways (mechanisms) responsible for URMC-099 facilitated nanoformulated antiretroviral responses in mononuclear phagocytes (MP; monocyte-derived macrophages [MDM] and microglia) in relation to MLKs and other related kinases targeted by URMC-099 (including kinases that mediate inflammation) and determine whether virus can be eradicated or merely suppressed. (2) To assess the impact of Rab family members identified in Aim 1 on potential drug toxicities from URMC-099 and nanoART in our models of HAND. To this end, we will investigate its roles in regulating MP and synaptic networks exposed to HIV-1 neurotoxins after URMC-099 and nanoART treatment. We will use in vivo models to further understand the role of Rabs as they relate to MLK activation and synaptodendritic damage with the idea of protecting the brain against further injuries. Finally, in (3) we will validate the utility of nanoformulated antiretrovral therapy (nanoART) and URMC-099 in clearance of persistently infected viral reservoirs in the CNS in our humanized CD34 engrafted model of neuroAIDS. Together, these aims will advance our approach to eradication of long-lived persistent infection of CNS MP while preserving synaptic architecture and function in disease.