INTERFERON SIGNAL ENHANCERS AS ANTIVIRAL THERAPEUTICS

描述（由申请人提供）：目前大多数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，包括新出现的病毒株。基于临床和临床前观察，我们提供了证据，证明干扰素（IFN）信号系统可以通过基因工程来提高效率，从而在体外和体内更好地抵抗RNA病毒感染，而且没有毒性。因此，我们继续鉴定与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，使用基于细胞的表型筛选来增加IFN刺激反应元件（ISRE）活性，以识别增强IFN信号通路的小分子。针对一系列已批准或批准的化合物的试点筛选提供了概念证明，产生了增强IFN信号通路和控制体外病毒水平的击中化合物。随后，我们筛选了另外两个化学文库，数据的高分辨率和可重复性使我们能够进行新的组合分析，最终选择并验证了75个已确认的命中。结构分析显示有11个簇，其中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强大的IFN增强和抗病毒活性（包括常见RNA呼吸道病毒），并且这种作用可能独立于3-羟基-3甲基戊二酰辅酶a （HMG-CoA）还原酶（HMGCR）抑制，这解释了为什么他汀类药物对固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信号的化合物将导致具有新作用机制的下一代抗病毒药物，并提出实现这一目标的具体目标如下：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传统作用。2. 确定IFN信号通路中他汀类药物抗病毒作用所需的靶点，进一步明确构效关系（SAR），进一步优化类似物选择。3. 定义并验证他汀类药物对细胞和动物RNA病毒感染的影响

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.