Novel Interferons and small molecule enhancers of the interferon pathway

IFNa的产生是病毒感染后宿主最早的免疫反应之一。这种I型干扰素的根本重要性在于它能够通过直接和间接的机制诱导先天和适应性免疫细胞的下游激活。由于I型干扰素具有强大的抗病毒活性，因此它们既可作为单一疗法使用，也可与其他小分子联合使用，以增强慢性病毒感染（如丙型肝炎）的治疗效果。尽管只有一部分患者对治疗有反应，而且长期给药方案的不良副作用是一个重大障碍，但干扰素仍然是最佳的治疗标准。因此，该项目的总体目标是并行开发两类IFNa增强剂，一种蛋白质和另一种小分子，可单独使用或与其他治疗药物联合使用，用于病毒感染的广谱治疗。首先，为了改善对ifn的功能反应并降低其毒性，Garcia小组开发了一种方法，用重组蛋白重新设计和扩展现有ifn的菜单，这些重组蛋白表现出优越的治疗特性。这种被称为体外进化的方法，已经被Garcia小组成功地用于设计具有独特结构-活性关系的细胞因子，并且已被证明比天然细胞因子更有效。其次，Khosia小组将对最近发现的一种聚酮天然产物a -74528进行化学生物学研究，该产物已被发现可以增强IFNa的抗病毒作用。Khosia集团还将从事A-74528类似物的生产和生化及结构分析。在目标1中，我们将利用体外进化来设计和改善IFN的抗病毒活性，以创建具有改变亲和力或与受体复合物二聚化几何形状的IFN。在目标2中，我们将对候选ifn进行生物物理研究，以确定其作为单一疗法或与其他疗法联合使用的作用机制。在Aim 3中，我们将通过动力学和剂量反应分析来研究A- 74528的抑制机制和药理学特性。在Aim 4中，我们将设计和合成A- 74528的类似物来定义结构活性关系。这种独特的蛋白质和小分子药物化学的结合有望产生高度颗粒化的、可预测的结构-活性指标，将IFN作用与抗病毒功能的分子和机制参数联系起来。

Production of IFNa is one of the earliest host immune responses after viral infection. The fundamental importance of this Type I interferon, lies in its ability to induce downstream activation of both innate and adaptive immune cells via direct and indirect mechanisms. Because they promote potent anti-viral activity, Type I IFNs have been used as both a monotherapy and in combination with other small molecules for enhanced efficacy in the treatment of chronic viral infections, such as Hepatitis C. Despite the fact that only a portion of patients respond to treatment, and that adverse side-effects from long term dosing regimens are a significant barrier, IFNs remain the best standard of care. Thus, it is the overall goal of this project to develop, in parallel, two classes of IFNa enhancers, one protein and the other small molecule, that may be used alone or in combination with other therapeutics for the broad-spectrum treatment of viral infections. First, in an effort to improve functional responses to IFNs and reduce their toxicity, the Garcia group has developed an approach to re-engineer and expand the menu of existing IFNs with recombinant proteins that exhibit superior therapeutic properties. This method, known as in vitro evolution, has been previously and successfully employed by the Garcia group to engineer cytokines with unique structure-activity relationships and that have proven to be more efficacious than the natural cytokine. Second, the Khosia group will pursue chemical biological studies on a recently discovered polyketide natural product, A-74528, that has been found to enhance the antirviral effects of IFNa. The Khosia group will also engage in production and biochemical and structural analysis of A-74528 analogs. In Aim 1 we will engineer and improve IFN antiviral activities using in vitro evolution to create IFNs with altered affinity or dimerization geometries with the receptor complex. In Aim 2 we will conduct biophysical studies of candidate IFNs to determine their mechanisms of action, as monotherapies or in combination with other therapeutics. In Aim 3 we will investigate the inhibitory mechanism and pharmacological properties of A- 74528 through kinetic and dose-response analyses. In Aim 4 we will design and synthesize analogs of A- 74528 to define structure activity relationships. This unique interfacing of protein and small molecule medicinal chemistry will hopefully yield highly granular, predictive structure-activity metrics linking the molecular and mechanistic parameters of IFN action to antiviral function.