Development of in vivo probes to study the function of TRIP8b in cognition

开发体内探针来研究 TRIP8b 在认知中的功能

• 批准号: 10665810
• 负责人: Dane M Chetkovich
• 金额: $ 82.5万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665810
• 关键词: Address; Animal Model; Arrhythmia; Binding; Biochemical; Biological; Biological Assay; Biology; Biophysics; Brain; Brain region; Cardiac; Cells; Cellular Assay; Characteristics; Chemicals; Chronic; Chronic stress; Cognition; Cognitive; Cognitive deficits; Cyclic Nucleotides; Data; Development; Dorsal; Drug Targeting; Electrophysiology (science); Flow Cytometry; Genetic; Heart; High Prevalence; Hippocampus; Impaired cognition; Impairment; In Vitro; Knock-out; Knowledge; Learning; Long-Term Potentiation; Major Depressive Disorder; Mediating; Memory; Mental Health; Mental disorders; Methods; Molecular; Molecular Target; Mus; Neurons; Nootropic Agents; Pathway interactions; Performance; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacotherapy; Phenotype; Play; Process; Property; Publishing; Reporting; Risk Factors; Role; Series; Specificity; Surface; Symptoms; Testing; Toxic effect; Translational Research; Validation; Work; analog; cognitive change; cognitive process; common symptom; cyclic-nucleotide gated ion channels; experimental study; high throughput screening; hippocampal atrophy; hippocampal pyramidal neuron; improved; in vivo; inhibitor; insight; interest; mouse model; neuronal excitability; neuropsychiatric disorder; novel; pharmacologic; reduce symptoms; response; scaffold; side effect; small molecule; small molecule inhibitor; social defeat; therapeutic target; therapy design; tool; trafficking; translational medicine; validation studies

Abstract Cognitive dysfunction is a common feature of neuropsychiatric diseases related to chronic stress. These symptoms can be debilitating, and despite the high prevalence of illnesses like Major Depressive Disorder, there are relatively few treatment options. To better address these symptoms, novel molecular targets are needed. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels play a critical role in regulating excitability in the hippocampus, a brain region critically involved in cognition. To avoid cardiac side effects associated with antagonizing HCN channels (which are also expressed in the heart), our group has focused on TRIP8b, a neuron-specific subunit of HCN channels. In our recently published report (Lyman et al, in press at Science Translational Medicine), we established that chemogenetically disrupting the interaction between TRIP8b and HCN in the dorsal hippocampus is sufficient to rescue impairments in cognition after chronic stress. Combined, these results are consistent with the hypothesis that a small molecule that disrupts the TRIP8b-HCN interaction could rescue the cognitive dysfunction seen after chronic stress. In order to capitalize on this discovery, our group recently executed a high throughput screen to identify small molecule inhibitors of the TRIP8b-HCN interaction. We identified one candidate scaffold which shows promising activity as a TRIP8b inhibitor. This compound series serves as the basis for our current submission in which we propose further medicinal chemistry optimization to develop TRIP8b inhibitors for use as chemical probes in order to study TRIP8b in vivo. In our enclosed preliminary data, we show that our inhibitor potently disrupts the interaction between TRIP8b and HCN in cells with negligible toxicity. This inhibition produces robust effects on Ih, the current mediated by HCN channels both in vivo and in vitro. To expand upon this work, we propose three aims. In Aim 1, we will perform iterative medicinal chemistry optimization to develop compounds with improved potency and pharmaceutical characteristics. Compounds that perform well in our biochemical and biophysical assays will be advanced to cellular assays in Aim 2 in order to determine if they are capable of disrupting the TRIP8b-HCN interaction. In Aim 3 we will utilize our compounds in vivo to study their effect on hippocampal function. Promising compounds will be investigated for their ability to rescue cognitive deficits that result from chronic social defeat, an animal model of chronic stress. These experiments will enable us to develop TRIP8b inhibitors that can be used as chemical probes to study the role of HCN channels in regulating cognitive dysfunction after chronic stress.

抽象的 认知功能障碍是与慢性应激相关的神经精神疾病的共同特征。这些 症状可能会使人衰弱，尽管重度抑郁症等疾病的患病率很高， 治疗选择相对较少。为了更好地解决这些症状，新的分子靶点是 需要。超极化激活的环核苷酸门控 (HCN) 通道在调节中发挥关键作用 海马体的兴奋性，海马体是与认知密切相关的大脑区域。避免心脏副作用 与拮抗 HCN 通道（也在心脏中表达）相关，我们小组重点关注 TRIP8b，HCN 通道的神经元特异性亚基。在我们最近发表的报告中（Lyman 等人， 科学转化医学出版社），我们确定化学遗传学破坏了相互作用 TRIP8b 和 HCN 之间的背侧海马足以挽救认知障碍 慢性压力。综合起来，这些结果与破坏小分子的假设是一致的。 TRIP8b-HCN 相互作用可以挽救慢性压力后出现的认知功能障碍。为了 利用这一发现，我们的团队最近进行了高通量筛选来识别小分子 TRIP8b-HCN 相互作用的抑制剂。我们确定了一种候选支架，其显示出有希望的活性 作为 TRIP8b 抑制剂。该化合物系列是我们目前提交的材料的基础，其中我们 提出进一步的药物化学优化，以开发 TRIP8b 抑制剂，用作化学探针 为了在体内研究 TRIP8b。在我们随附的初步数据中，我们表明我们的抑制剂可以有效破坏 TRIP8b 和 HCN 在细胞中的相互作用，毒性可忽略不计。这种抑制产生强大的 对体内和体外 HCN 通道介导的电流 Ih 的影响。为了扩展这项工作，我们 提出三个目标。在目标 1 中，我们将进行迭代药物化学优化来开发 具有改进的效力和药物特性的化合物。在我们的产品中表现良好的化合物 目标 2 中的生化和生物物理测定将升级为细胞测定，以确定它们是否 能够破坏 TRIP8b-HCN 相互作用。在目标 3 中，我们将利用我们的化合物进行体内研究 它们对海马功能的影响。将研究有前途的化合物的救援能力 由于慢性社会失败而导致的认知缺陷，这是慢性压力的动物模型。这些实验 将使我们能够开发 TRIP8b 抑制剂，可用作化学探针来研究 HCN 的作用 慢性应激后调节认知功能障碍的通道。