Project 2: PK/PD requirements for mucolytic therapeutic agents in vitro and in vivo

项目2：粘液溶解治疗剂体内外PK/PD要求

基本信息

批准号：
10001600
负责人：
Richard Charles Boucher
金额：
$ 47.8万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-07 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10001600
关键词：
Acetates Adhesions Adhesives Aerosols Animals Ascaris suum Asthma Back Basic Science Biological Models Body Weight decreased Cell Culture Techniques Chronic Obstructive Airway Disease Cleaved cell Clinic Clinical Clinical Data Collaborations Data Development Disease Dose Drug Delivery Systems Environment Epithelial Cells Equilibrium Exhibits Exposure to Focal Adhesions Formulation Frequencies Gel Generations Goals Health Human Hydration status In Vitro Infection Inflammation Inhalation Interleukin-1 beta Interleukin-13 Lead Liquid substance Lung Lung diseases MUC5AC gene MUC5B gene Measures Modeling Modification Molecular Weight Mucins Mucolytics Mucous body substance Mus Oxidation-Reduction Oxidative Stress Parents Patients Pharmaceutical Preparations Phenotype Polymers Population Property Public Health Pyroglyphidae Reducing Agents Rehydrations Research Personnel Resistance Role Safety Saline Sheep Speed Sulfhydryl Compounds Surface System Testing Therapeutic Therapeutic Agents Toxicology Translating United States airway inflammation airway obstruction airway remodeling asthma model asthmatic base biophysical analysis bronchial epithelium clinical candidate crosslink data modeling design dithiol effective therapy esterase extracellular flexibility in vivo in vivo Model inflammatory lung disease model design mouse model mucus clearance mucus-associated lung diseases novel novel therapeutics pharmacokinetics and pharmacodynamics preclinical development programs relative effectiveness response tool

项目摘要

Project 2 has focused on the hypotheses that: (1) hyperconcentrated (dehydrated) mucus produces airway mucus adhesion/plaques that drive the progression of muco-obstructive lung diseases; and, (2) that clearance of these plugs/plaques will be therapeutically useful in patients with the muco-obstructive phenotype. Recent data have indicated that once mucostasis has occurred, modification of mucus plaques may occur that make them “permanent” and resistant to rehydration therapies. Our biophysical analyses suggest that mucin molecular weight (MW) reduction will be effective in clearing mucus in this setting without rehydration. Further, Project 2 hypothesizes that by the generation of disease specific models, that effectively mimic the extracellular mucins/mucus targets of our disease populations, will produce useful information to guide the clinical projects with respect to selection of drug doses, dosing frequency, requirement for an active vehicle (e.g., hypertonic saline) and generate data on “off target” redox effects. In collaboration with tPPG investigators, we have established that in CF (and COPD) MUC5B is the dominant mucin obstructing airways. In contrast, we have identified that MUC5AC is the dominant airway mucin in subjects with asthma. Accordingly, we have generated appropriate cell culture, small animal, and sheep models that mimic the mucin dominance phenotype for each disease entity. Utilizing these tools, Project 2 will measure and compare the pharmacodynamics and pharmacokinetic properties of two different, but complementary, thiol reducing agent clinical candidates. P2176 is a di-thiol compound with rapid reductive activity (Kcat) whereas the mono-thiol P2114 has a slower intrinsic reductive (Kcat) activity. Both molecules share features designed to retain the molecules on the airway surface and both molecules have their thiols “capped” by acetate groups that are cleaved by esterases on airway surfaces to liberate active compounds. The relative effectiveness/properties of each molecule will be tested in: (1) in vitro HBE cultures designed to mimic the extracellular MUC5AC/MUC5B ratios of CF/COPD (Il-1β treated) and asthma (IL-13 treated); (2) mouse models of CF/COPD (βENaC) mice, and asthma (house dust mite treated) to measure clearance of mucus plugs after aerosol drug delivery; and, large animal sheep models, including wild-type, Ascaris suum exposed, and HNE/CF172 treated sheep to measure pharmacodynamics and pharmacokinetic parameters. Based on these data, and GLP toxicology studies, a lead molecule doses/dosing frequency, and a vehicle will be selected. Importantly, Project 2 will also utilize information gained from Project 1/Project 2 studies and clinical Projects 3 and 4 to study backup compounds as improvements in the frontrunner become identified/implemented in the parent thiol compound chassis. Thus the overarching goal of the 2b Project is to identify the optimal drug properties, balancing rate of mucin thiol reduction, duration of activity, and safety, to develop a new class of muco-clearance assisting agents that focus on mucin MW reduction that can be delivered alone or in combination with hydrating agents.

项目 2 重点关注以下假设：(1) 超浓缩（脱水）粘液产生气道导致粘液阻塞性肺部疾病进展的粘液粘附/斑块，以及（2）清除；这些栓塞/斑块对于患有粘液阻塞表型的患者具有治疗作用。数据表明，一旦发生粘膜停滞，粘液斑块可能会发生改变，从而使我们的生物物理分析表明，它们是“永久性的”并且对补液疗法具有抵抗力。在这种情况下，分子量（MW）的降低将有效地清除粘液，而无需补液。项目 2 强调，通过生成疾病特定模型，可以有效地模仿我们疾病人群的细胞外粘蛋白/粘液目标，将产生有用的信息来指导关于药物剂量选择、给药频率、主动载体要求的临床项目（例如，高渗盐水）并与 tPPG 合作生成“脱靶”氧化还原效应的数据。研究人员发现，在 CF（和 COPD）中，MUC5B 是阻塞气道的主要粘蛋白。相比之下，我们已经确定 MUC5AC 是哮喘受试者中的主要气道粘蛋白。我们已经生成了模仿粘蛋白的适当细胞培养物、小动物和绵羊模型利用这些工具，项目 2 将测量和比较每种疾病实体的显性表型。两种不同但互补的硫醇还原剂的药效学和药代动力学特性 P2176 是一种具有快速还原活性 (Kcat) 的二硫醇化合物，而单硫醇化合物。 P2114 具有较慢的固有还原 (Kcat) 活性，这两种分子具有旨在保留 P2114 的共同特征。气道表面上的分子，并且两个分子的硫醇都被乙酸酯基团“封端” 被气道表面的酯酶裂解以释放活性化合物的相对有效性/特性。每个分子将在以下环境中进行测试：(1) 旨在模拟细胞外 MUC5AC/MUC5B 的体外 HBE 培养物 CF/COPD（IL-1β治疗）和哮喘（IL-13治疗）的比率；（2）CF/COPD（βENaC）小鼠模型，和哮喘（经过屋尘螨处理）以测量气雾剂药物输送后粘液塞的清除率；大型动物绵羊模型，包括野生型、暴露的猪蛔虫和 HNE/CF172 处理的绵羊根据这些数据和 GLP 毒理学测量药效学和药代动力学参数。重要的是，项目 2 还将选择研究、先导分子剂量/给药频率和载体。利用从项目 1/项目 2 研究以及临床项目 3 和 4 获得的信息来研究备份作为领先者改进的化合物在母体硫醇化合物中得到识别/实现因此，2b 项目的总体目标是确定最佳药物特性、平衡速率。粘蛋白硫醇减少、活性持续时间和安全性，以开发一类新的粘膜清除辅助剂专注于降低粘蛋白分子量的药剂，可以单独使用或与保湿剂组合使用。