Compound/Combination Selection Core

慢性阻塞性肺疾病和慢性阻塞性肺疾病的发病机制是多因素的，但 包括一个常见的致病步骤：粘液纤毛的减少 净空(MCC)。基于形成TPPG基础的研究 项目I和II，已经观察到正常的粘液清除 需要充分的呼吸道表面水合作用(见计划 概述)。然而，慢性阻塞性肺疾病和慢性阻塞性肺疾病患者都表现出增加的 呼吸道粘液浓度(固体百分比)，因呼吸道减少所致 由于CFTR功能障碍(1)、粘蛋白高分泌(2)或 两者的结合。其结果是出现了呼吸道粘附物 粘液斑块与纤毛和咳嗽依赖粘液的减慢 清除，导致慢性呼吸道感染。因此，发展 加强呼吸道粘液清除的治疗方法可能会 使慢性阻塞性肺疾病/慢性阻塞性肺病患者受益。Core E的主要目标是支持所有 TPPG的项目通过确定最有效的治疗方法 增粘剂，增强粘液的粘附性和传输性。 作为这个tPPG项目的一部分，我们已经确定了许多类 候选治疗药物，预计会改变 粘液，促进其清除。这些潜在的类别和示例 代理汇总如表1所示。 因为粘液的粘弹性是浓度依赖的，所以试剂 哪些会增加粘液的水合状态(称为水化器) 研究(3)。这将包括渗透激素(如高渗盐水和 甘露醇)以及ENaC介导的体液重吸收的改进剂， 包括：ENAC阻滞剂(Parion P552和PI 527)和ENaC的修饰剂 活动(QUB和Splunk)。作为减少粘液的另一种方法 浓缩，我们还将评估大环内酯类的使用，这些大环内酯类药物已经 显示减少粘液表达(4)。除了阿奇霉素，我们还将测试 来自Cempra和Gilead的一系列新的大环内酯类药物 优化以提高他们的活跃度(请参阅支持函)。 除了降低粘液浓度外，我们还将寻求 确定降低其粘弹性和粘附性的试剂，方法是改变 粘蛋白-粘蛋白和粘蛋白-蛋白质的相互作用。二硫基还原剂 预计会通过降低粘液的粘弹性来降低粘弹性 粘蛋白单体头尾相连的链状结构。除了现有的减少 试剂(例如N-乙酰半胱氨酸和DTT)我们还将测试新的还原 干扰素诱导的溶酶体硫醇还原酶等药物 (Gilt)来自耶鲁大学的Peter Cresswell博士和各种新颖的重组- Parion的DTT(DTT-R)可提高效率和持续时间 行动(请参阅计划简介)。表面活性剂和洗涤剂会破坏 粘液层内的蛋白质-蛋白质相互作用被设想为 降低粘液的粘附性(5-6)。有很多种 其他假定的粘液改变剂(例如，钙离子螯合剂(7)，葡聚糖(8)， 肝素(9)、碳酸氢钠(10))，我们将进行研究，以确定恢复/刺激MCC的新疗法。 核心E由三个不同的职能部门组成，它们将确定潜在用于 人体临床试验。此核心的总体战略及其与每个tPPG项目的互动显示在 图1.总体概念是使用体外粘液筛选来筛选大量测试试剂 方法并确定在后续细胞培养和小鼠研究中最有效的试剂(S) 学习。粘液流变性和水化核心服务的主要目标是提供项目I和II 调查人员对所有感兴趣的药物进行全面分析，并建立剂量-效应关系 关于(1)粘液粘弹性的变化--粘液流动性的一种度量，以及(2)粘液的水化 气道口。一旦确定了最有效的候选代理，每个代理对 刺激和恢复人呼吸道上皮细胞粘液运输的研究将在下一个核心中进行 服务。最后，在第三个核心部分，将测试以前研究中最有效的药剂 在单剂量和多剂量测试方案期间用于阻塞性呼吸道疾病的小鼠模型。其目的是评估药物的疗效和毒性，如通过组织病理学、形态测量、 生化分析和炎性细胞计数。一旦确定，这些代理(或代理类别)将成为 项目III中人体试验的重点。

The pathogenesis of CF and COPD lung disease is multi-factorial, but includes a common disease-initiating step: the reduction of mucociliary clearance (MCC). Based on studies shaping the foundation of the tPPG Projects I and II, it has been observed that normal mucus clearance requires adequate hydration of the airway surface (see Program Overview). However, both CF and COPD patients exhibit an increase in airway mucus concentration (% solids), as a result of reduced airway hydration, due to CFTR dysfunction (1), mucin hypersecretion (2), or a combination of the two. The result is the appearance of airway-adherent mucus plaques and the slowing of both cilial- and cough-dependent mucus clearance, leading to chronic airway infections. Therefore, developing therapies to enhance the clearance of mucus from the airways is likely to benefit patients with CF/COPD. The primary goal of Core E is to support all the Projects of the tPPG by identifying the most effective therapeutic agents to enhance the dis-adherence and transportability of mucus. As part of this tPPG project, we have identified a number of classes of candidate therapeutic agents, which are predicted to alter the properties of mucus, facilitating its clearance. These classes and examples of potential agents are summarized in Table 1. Because the viscoelasticity of mucus is concentration dependant, agents which increase the hydration state of mucus (called hydrators) will be studied (3). This will include osmolytes (such as hypertonic saline and mannitol) as well as modifiers of ENaC-mediated fluid reabsorption, including: ENaC blockers (Parion P552 and PI 527) and modifiers of ENaC activity (QUB and SPLUNK). As an alternative approach to reduce mucus concentration, we will also evaluate the use of macrolides, which have been shown to reduce mucus expression (4). As well as Azithromycin, we will test a series of novel macrolide agents from Cempra and Gilead which are optimized to improve their activity (see Letters of Support). In addition to lowering the concentration of mucus, we will also seek to identify agents which reduce its viscoelasticity and adhesivity, by altering mucin-mucin and mucin-protein interactions. Di-sulfhydryl reducing agents are predicted to decrease the viscoelasticity of mucus by decreasing the head-to-tail chaining of mucin monomers. In addition to existing reducing agents (e.g. N-acetylcysteine and DTT) we will also test novel reducing agents such as Gamma-interferon-inducible lysosomal thiol reductase (GILT) from Dr. Peter Cresswell (Yale) and a variety of novel recombinant- DTTs (DTT-Rs) from Parion which increase the efficiency and duration of action (see Program Introduction). Surfactants and detergents that disrupt protein-protein interactions within the mucus layer are envisaged to decrease the cohesive properties of mucus (5-6). There are a number of other putative mucus-altering agents (e.g. Ca2+ chelators (7), dextran (8), heparin (9), bicarbonate (10)) that we will study to identify novel therapeutics for restoring/ stimulating MCC. Core E consists of three distinct functional divisions that will identify effective agents for potential use in a human clinical trial. The overall strategy of this Core and its interactions with each tPPG Project is shown in Figure 1. The general concept is to screen a large number of test agents using in vitro mucus screening methodologies and identify the most effective agent(s) to be studied in subsequent cell culture and mouse studies. The primary goal of the mucus rheology and hydration core service is to provide Project I & II investigators with a comprehensive analysis of all agents of interest and establish the dose-effect relationship on changes in (1) mucus viscoelasticity - a measure of the "flowability" the mucus, and (2) the hydration of the airway. Once the most effective candidate agents are identified, the effect of each of these agents on stimulating and restoring mucus transport in human airway epithelial cultures will be studied in the next core service. Finally, in the third core component, the most effective agents from the previous studies will be tested in a mouse model for obstructive airway diseases during a single and multiple dose testing regimens. The goal is to evaluate drug efficacy and toxicity, as assessed by histopathology, morphometric measurements, biochemical analysis and inflammatory cell count. Once identified, these agents (or class of agents) will be the focus of a human trial in Project III.