Identification of small molecules for neurological complications of HIV and substance abuse comorbidity

鉴定治疗 HIV 神经并发症和药物滥用合并症的小分子

• 批准号: 10343707
• 负责人: PIETRO P SANNA
• 金额: $ 76.69万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10343707
• 关键词: Affect; Alcohol abuse; Alcohols; Algorithmic Analysis; Animal Model; Biogenesis; Biological Assay; Biological Availability; Brain; California; Cell Line; Cells; Chemicals; Dependence; Disease; Down-Regulation; Drug Kinetics; Florida; Gene Expression; Gene Expression Profiling; Gene set enrichment analysis; Genes; Glutamates; HIV; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Hippocampus (Brain); Human; Impaired cognition; Impairment; In Vitro; Individual; Injury; Investigation; Laboratories; Lead; Libraries; Mediating; Mediator of activation protein; Memory; Mitochondria; Modeling; Molecular; Nature; Nerve Degeneration; Neurobiology; Neurologic; Neuronal Dysfunction; Neuronal Injury; Neurons; Oral; Oxidative Stress; PC12 Cells; Patients; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phenotype; Polymerase Chain Reaction; Population; Prefrontal Cortex; Property; Rattus; Regulator Genes; Reporter; Research; Research Institute; Rodent; Rodent Model; Self Administration; Series; Source; Spices; Structure-Activity Relationship; Substance abuse problem; Synapses; System; Systems Biology; Testing; Transgenic Organisms; Triage; Validation; analog; antiretroviral therapy; base; cell type; comorbidity; counterscreen; cytotoxicity; drug discovery; drug metabolism; effective therapy; efficacy testing; executive function; frontal lobe; genetic signature; high throughput screening; in vivo; in vivo evaluation; innovation; lead optimization; lead series; luminescence; methamphetamine abuse; neuroAIDS; neuronal survival; physical property; prevent; programs; promoter; receptor; resilience; response; scaffold; scale up; screening; skills; small molecule; therapeutic target; transcription factor; transcriptome sequencing; translational medicine

Summary HIV-associated neurocognitive disorders (HANDs) and substance abuse comorbidity remain prevalent despite combination antiretroviral therapy (cART). The innovative hypothesis behind this project is that reversing trophic deficits that characterize HANDs and manifest as synaptodendritic and mitochondrial injury can effectively prevent or ameliorate cognitive decline in HIV and compulsive drug taking. To test this hypothesis, we will perform a throughput screening campaign (HTS) to identify modulators of a newly identified gene regulatory mechanism that broadly regulates trophic support and mitochondrial biogenesis in key cortical and hippocampal neuronal populations and that we found are downregulated in both neuroAIDS and dependence. We expect that small molecules that are able to upregulate this mechanism will make neurons more resilient to oxidative stress and neurodegeneration, resulting in a reduction of neuronal injury and hypofrontality, which is a key cause of cognitive dysfunction in HIV and compulsive drug and alcohol taking in dependent individuals. We will employ cell-based luminescence reporter assays in a 1536-well plate format in conjunction with a tiered approach to screen the >640,000-compound Scripps Drug Discovery Library (SDDL) and confirm the potency of ~500 drug-like molecules. Hit-validation will be performed to identify small-molecule regulators and eliminate nonspecific effectors using parallel orthogonal assays and off-target assessment using cytotoxicity counterscreens employing disease-relevant cell types. To prioritize hit scaffold series, we will select analogs of confirmed hits from both the existing compound libraries and other commercial sources. Hit scaffolds will be triaged to remove intractable molecules. We will select 3-5 molecular series from the most promising hits which, will be profiled to verify the selectivity, potency, and lack of cytotoxicity. Using an orchestrated effort from the applicant laboratories (Scripps California and Scripps Florida), leads in 2-4 series will be formulated and retested for potency/selectivity with the aim of advancing leads that can elicit the appropriate in vitro response in the aforementioned assays and can be evaluated for appropriate phenotypic responses to cell-expression levels. This will be followed by in vitro and in vivo pharmacokinetics (PK) studies to identify 1-2 top scaffolds for further investigation. Finally, the most promising 2-3 compounds with favorable drug metabolism and pharmacokinetics (DMPK) properties will be selected for in vivo testing, including high oral bioavailability, and will be scaled up and tested in vivo in rodent efficacy models. Efficacy in regulating the expression of the gene expression program under study will be validated by quantitative polymerase chain reaction of synaptic, dendritic, and mitochondrial genes and RNA-Seq in conjunction with Gene Set Enrichment Analyses (GSEA). We will then test the ability of the optimized probe compounds to prevent or ameliorate cognitive impairment that is attributable to prefrontal and hippocampal degenerative changes in HIV-1 transgenic rats with and without concomitant compulsive drug or alcohol self-administration.

总结 HIV相关的神经认知障碍（HANDs）和药物滥用合并症仍然普遍存在 尽管联合抗逆转录病毒治疗（cART）。这个项目背后的创新假设是， HANDs的营养缺陷表现为突触树突和线粒体损伤， 有效预防或改善艾滋病认知能力下降和强迫性吸毒。为了检验这一假设， 我们将进行一项通量筛选活动（HTS），以确定一个新发现的基因的调节剂 广泛调节关键皮层和皮层中营养支持和线粒体生物发生的调节机制， 海马神经元群体，我们发现在neuroAIDS和依赖中下调。 我们预计能够上调这种机制的小分子将使神经元更有弹性， 氧化应激和神经变性，导致神经元损伤和额叶功能减退，这是一个 艾滋病患者认知功能障碍的主要原因以及依赖性个体的强迫性药物和酒精摄入。我们 将在1536孔板格式中采用基于细胞的发光报告基因测定， 筛选超过640，000种化合物的Scripps药物发现库（SDDL）并确认 ~500个药物样分子。将进行命中验证以鉴定小分子调节剂并消除 使用平行正交试验的非特异性效应物和使用细胞毒性的脱靶评估 使用疾病相关细胞类型的反筛选。为了优先考虑命中支架系列，我们将选择类似物 从现有的化合物库和其他商业来源确认命中。打脚手架将是 进行了分流以去除难处理的分子我们将从最有希望的命中中选择3-5个分子系列， 将进行分析，以验证选择性、效力和无细胞毒性。通过精心策划， 申请实验室（Scripps加州和Scripps佛罗里达），将配制2-4个系列的电极导线并进行重新检测 用于效力/选择性，目的是推进可在体外引起适当反应的先导物， 这些表型反应可以通过上述测定来进行，并且可以评估对细胞表达水平的适当表型反应。 随后将进行体外和体内药代动力学（PK）研究，以确定1-2种最佳支架，用于进一步研究。 调查最后，最有希望的2-3个化合物具有有利的药物代谢和药代动力学 将选择DMPK的性质用于体内测试，包括高口服生物利用度，并将按比例放大， 在啮齿动物功效模型中进行体内测试。调节基因表达程序表达的功效 将通过突触、树突和线粒体的定量聚合酶链反应进行验证。 基因和RNA-Seq结合基因集富集分析（GSEA）。然后我们将测试 所述优化的探针化合物用于预防或改善可归因于前额叶的认知损害 HIV-1转基因大鼠在伴随和不伴随强迫性药物的情况下的海马退行性改变 或酒精自我管理。