INTERFERON SIGNAL ENHANCERS AS ANTIVIRAL THERAPEUTICS

干扰素信号增强剂作为抗病毒治疗

• 批准号: 8697863
• 负责人: Michael J Holtzman
• 金额: $ 44.09万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8697863
• 关键词: Animal Model; Antiviral Agents; Biological Assay; Breathing; Cell Culture System; Cell model; Cells; Clinical; Coenzyme A; Collection; Communicable Diseases; Data; Dependency; Development; Drug Design; Effectiveness; Enhancers; Epidemic; Epithelial Cells; Genetic Engineering; Goals; Human; Hydroxymethylglutaryl-CoA reductase; Immune response; In Vitro; Individual; Infection; Interferons; Lead; Ligands; Mediating; Null Lymphocytes; Oxidoreductase; Pathway interactions; Pharmacology; Property; Protein Binding; Proteins; Proteomics; RNA; RNA Virus Infections; RNA Viruses; Reproducibility; Resistance; Resolution; Respiratory Tract Infections; Response Elements; Role; STAT1 gene; Safety; Signal Pathway; Signal Transduction; Small Interfering RNA; Sterols; Structure; Structure-Activity Relationship; System; Testing; Therapeutic; Toxic effect; Validation; Viral; Viral Genome; Virus; Virus Diseases; analog; base; drug development; drug discovery; high throughput screening; in vivo; mouse model; next generation; novel; pathogen; pharmacophore; pre-clinical; prevent; public health relevance; research study; respiratory virus; safety study; small molecule; small molecule libraries; therapeutic target

DESCRIPTION (provided by applicant): Most current approaches for antiviral therapeutics target the virus specifically and directly, but that strategy can be difficult in the face of a rapdly evolving viral genome and consequent pathogen resistance. An alternative approach would be to enhance the host innate immune response, which should prove efficacious to a broad range of viruses, including new and emergent strains. Based on clinical and pre-clinical observations, we provided evidence that the interferon (IFN) signaling system could be genetically engineered for increased efficiency and thereby better protect against RNA virus infection in vitro and in viv without toxicity. We therefore proceeded to identify small molecules that interact with the IFN signaling system to mimic this antiviral benefit and thereby provide therapeutic leads for further development. We developed a high- throughput screening (HTS) strategy using a cell-based phenotypic screen for increased IFN-stimulated response element (ISRE) activity to identify small molecules that enhance the IFN signaling pathway. A pilot screen against a collection of already approved or approvable compounds provided proof-of-concept by yielding hit compounds that enhanced the IFN signaling pathway and controlled viral level in vitro. We subsequently screened two other chemical libraries, and the high resolution and reproducibility of the data allowed for a novel combined analysis with the eventual selection and validation of 75 confirmed hits. Structural analysis showed 11 clusters, including one containing several statins. Functional analysis showed that statins indeed possess potent IFN enhancing and antiviral activity (including common RNA respiratory viruses), and this action may be independent of 3-hydroxy-3methylglutaryl-CoA (HMG-CoA) reductase (HMGCR) inhibition to explain why statin potency on sterol synthesis does not correlate with antiviral potency. We therefore hypothesize that developing compounds to enhance endogenous IFN signaling will lead to next- generation antiviral drugs with a novel mechanism of action and propose to achieve this goal with the following specific aims: 1. Determine whether the IFN-enhancing property of statins is independent of their conventional role in blocking the sterol synthesis pathway. 2. Determine the target of statins within the IFN signaling pathway that is required for an antiviral effect to further define structure-activity relationship (SAR) and further optimize analog selection. 3. Define and validate statin effect on RNA virus infection in cell and in animal models. 4. Use our established drug discovery and development system to discover and develop additional types of compounds with IFN signal enhancing and antiviral activity. We expect these studies to provide pre-clinical candidates for use as next-generation antiviral therapeutics, particularly targeting common RNA respiratory viruses.

描述（由申请人提供）：大多数目前的抗病毒治疗方法特异性和直接靶向病毒，但面对快速进化的病毒基因组和随之而来的病原体耐药性，这种策略可能很困难。另一种方法是增强宿主的先天免疫反应，这应该证明对广泛的病毒有效，包括新的和新出现的毒株。基于临床和临床前的观察，我们提供了证据表明，干扰素（IFN）信号系统可以通过基因工程提高效率，从而更好地防止RNA病毒感染在体外和体内没有毒性。因此，我们着手鉴定与IFN信号系统相互作用的小分子，以模拟这种抗病毒益处，从而为进一步开发提供治疗线索。我们开发了一种高通量筛选（HTS）策略，其使用基于细胞的表型筛选来增加IFN刺激的应答元件（ISRE）活性，以鉴定增强IFN信号传导途径的小分子。针对一系列已经批准或可批准的化合物进行的中试筛选通过产生增强干扰素信号通路并控制体外病毒水平的热门化合物来提供概念验证。我们随后筛选了另外两个化学文库，并且数据的高分辨率和可重复性允许进行新的组合分析，最终选择并验证75个确认的命中。结构分析显示了11个簇，包括一个含有几种他汀类药物。功能分析表明，他汀类药物确实具有有效的IFN增强和抗病毒活性（包括常见的RNA呼吸道病毒），这种作用可能不依赖于3-羟基-3甲基戊二酰辅酶A（HMG-CoA）还原酶（HMGCR）抑制，以解释为什么他汀类药物对甾醇合成的效力与抗病毒效力无关。因此，我们假设开发增强内源性IFN信号传导的化合物将导致具有新的作用机制的下一代抗病毒药物，并提出以以下具体目标实现该目标：1.确定他汀类药物的IFN增强特性是否独立于其阻断固醇合成途径的常规作用。2.确定他汀类药物在IFN信号通路中的靶点，这是抗病毒作用所需的，以进一步确定构效关系（SAR）并进一步优化类似物选择。3.确定和验证他汀类药物对细胞和动物中RNA病毒感染的作用 模型4.使用我们已建立的药物发现和开发系统来发现和开发具有IFN信号增强和抗病毒活性的其他类型的化合物。我们希望这些研究能够为下一代抗病毒治疗提供临床前候选药物，特别是针对常见的RNA呼吸道病毒。