Development of a Neutrophil Degranulation Inhibitor to Treat ARDS

开发治疗 ARDS 的中性粒细胞脱颗粒抑制剂

基本信息

批准号：
10697442
负责人：
Kenneth R MCLEISH
金额：
$ 30.64万
依托单位：
DEGRANIN THERAPEUTICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10697442
关键词：
Accounting Acute Lung Injury Acute Respiratory Distress Syndrome Admission activity Animal Model Animal Organ Animals Anti-Bacterial Agents Attenuated Bacteria Binding Biodistribution Biological Blood COVID-19 COVID-19 pandemic Cells Characteristics Clinical Clinical Trials Complication Critical Illness Cytoplasmic Granules Data Development Discipline of Nursing Disease Endotoxins Event Exocytosis Exocytosis Inhibition Extravasation Failure Family suidae Foundations Healthcare Systems Histopathologic Grade Host Defense Human Immune Immunohistochemistry In Vitro Inflammatory Influenza A Virus, H1N1 Subtype Inhalation Injury Innovative Therapy Intensive Care Units Laboratories Legal patent Life Limulus Lung Lung infections Maintenance Mass Spectrum Analysis Measures Modeling Molecular Target Mus Neutrophil Activation Neutrophil Infiltration Organ Oryctolagus cuniculus Outcome Pathogenesis Patients Peptides Permeability Phagocytes Phagocytosis Phagosomes Pharmaceutical Preparations Pharmacologic Substance Pharmacological Treatment Pharmacotherapy Phase Physicians Physiological Plasma Play Positron-Emission Tomography Production Proteins Protocols documentation Pulmonology Quality Control Radiolabeled Reactive Oxygen Species Recombinant Proteins Reproducibility Respiratory Failure Respiratory physiology Rodent Model Role SARS-CoV-2 infection SNAP receptor SNAP23 gene Scientist Sheep Small Business Innovation Research Grant Supportive care Syndrome Technology Therapeutic Therapeutic Effect Time Toxic Neutrophil Toxic effect Transmembrane Transport Treatment Cost Treatment Efficacy Validation Vascular Endothelial Cell Virus Work alveolar epithelium aptamer cecal ligation puncture cell injury clinically relevant commercialization cost drug candidate experimental study extracellular immunogenicity improved in vivo influenzavirus inhibitor innovation lung injury manufacturing capabilities manufacturing quality manufacturing run mortality mouse model neutrophil novel therapeutic intervention pharmacokinetics and pharmacodynamics pharmacologic phase 2 study pneumonia model preclinical study prevent recruit screening sepsis induced ARDS side effect therapeutic candidate therapeutic target

项目摘要

Project Summary Acute respiratory distress syndrome (ARDS) is a critical problem in pulmonary medicine, accounting for 10% of intensive care unit (ICU) admissions and totaling over 200,000 patients/year in the U.S. Current treatment cost typically exceeds $70,000 per patient. Decades of clinical trials failed to identify effective pharmacologic therapy for ARDS, while mortality remains above 30%. Thus, there is a critical, unmet clinical need for successful pharmacologic strategies to treat ARDS. Neutrophils play an essential role in the lung injury leading to ARDS, including that due to COVID-19, through extracellular release of reactive oxygen species (ROS), granule constituents, and neutrophil extracellular traps (NETs). The scientific foundation underlying our innovative therapy is that inhibition of neutrophil degranulation also prevents release of ROS and NETs, making degranulation a therapeutic target. We generated a recombinant protein, degranin-23 (degranulation inhibitor of SNAP-23, DGN-23) containing a SNARE motif from SNAP-23 and a cell permeability peptide. DGN-23 rapidly enters human neutrophils in vitro and in vivo, inhibits degranulation by 50% to 80%, prevents priming of ROS release, and reduces NET formation. Importantly, in vitro studies show DGN-23 does not impair neutrophil phagocytosis, granule fusion with phagosomes, or bacterial killing within phagosomes, and in vivo studies show that DGN-23 administration at the initiation of lung injury inhibits acute lung injury in 3 rodent models, without obvious toxicity. Thus, in vitro and in vivo data provide strong support for our novel therapeutic strategy that inhibition of neutrophil exocytosis attenuates lung injury leading to ARDS. The next steps are development of manufacturing capability and quality control to move from a laboratory grade recombinant protein to a pharmaceutical grade drug and validation of therapeutic effect without toxicity under clinically relevant conditions. This transition will be accomplished by two Aims. Aim 1: Characterize production characteristics and off-target/side effects of DGN-23. This aim will determine purity, presence of bacterial contaminants, yield, reproducibility of potency, and stability in multiple production runs of DGN-23. Cell toxicity and off target effects on circulating immune cells will be determined. Organ and cell localization of DGN-23 will be determined in all major organs from animals undergoing experiments described in Aim 2. Aim 2: Determine therapeutic efficacy of DGN-23 under clinically relevant conditions. The ability of DGN-23 to prevent acute lung injury and improve mortality when administered at various times after the initiating injury will be determined in a mouse model of H1N1 influenza virus-induced acute lung injury. Improved survival and improved blood oxygenation will serve as outcomes to proceed with development. At the conclusion of this work, this Phase 1 project will have identified a drug candidate with an optimal combination of efficacy, yield, stability, and potency to advance to advanced pre-clinical studies in a Phase 2 project.

项目概要急性呼吸窘迫综合征（ARDS）是肺部医学的一个关键问题，美国每年有 10% 的重症监护病房 (ICU) 入院患者，总计超过 200,000 名患者目前每位患者的治疗费用通常超过 70,000 美元。数十年的临床试验未能确定有效的 ARDS 的药物治疗，而死亡率仍高于 30%。因此，存在一个关键的、未满足的临床需要成功的药物策略来治疗 ARDS。中性粒细胞在肺中发挥重要作用通过细胞外释放活性氧而导致 ARDS 的损伤，包括由 COVID-19 引起的损伤种（ROS）、颗粒成分和中性粒细胞胞外陷阱（NET）。科学基础我们创新疗法的基础是抑制中性粒细胞脱颗粒也可以防止 ROS 的释放和 NET，使脱粒成为治疗目标。我们生成了重组蛋白 Degranin-23 （SNAP-23、DGN-23 的脱颗粒抑制剂）含有来自 SNAP-23 的 SNARE 基序和细胞渗透性肽。 DGN-23 在体外和体内快速进入人中性粒细胞，通过抑制脱颗粒 50% 至 80%，防止 ROS 释放的启动，并减少 NET 的形成。重要的是，体外研究表明 DGN-23 不会损害中性粒细胞的吞噬作用、颗粒与吞噬体的融合或内部的细菌杀灭作用吞噬体和体内研究表明，在肺损伤开始时给予 DGN-23 可抑制急性肺损伤。 3种啮齿类动物模型均出现肺损伤，未见明显毒性。因此，体外和体内数据为我们的新治疗策略是抑制中性粒细胞胞吐作用，减轻导致 ARDS 的肺损伤。下一步是从实验室发展制造能力和质量控制将重组蛋白分级为医药级药物并验证无毒性的治疗效果在临床相关条件下。这一转变将通过两个目标来实现。目标 1：表征 DGN-23 的生产特性和脱靶/副作用。这个目标将决定纯度、存在 DGN-23 多次生产运行中的细菌污染物、产量、效力再现性和稳定性。将确定细胞毒性和对循环免疫细胞的脱靶效应。器官和细胞定位将在进行目标 2 中描述的实验的动物的所有主要器官中测定 DGN-23。图2：确定DGN-23在临床相关条件下的治疗效果。 DGN-23的能力在损伤发生后的不同时间给药可预防急性肺损伤并降低死亡率将在 H1N1 流感病毒引起的急性肺损伤小鼠模型中进行测定。提高生存率并血液氧合的改善将成为发育的结果。在此结论中工作中，该第一阶段项目将确定一种具有功效、产量、稳定性和在第二阶段项目中推进高级临床前研究的潜力。