PARP Inhibitor and Redox Catalyst for Ventilatory Trauma

用于通气创伤的 PARP 抑制剂和氧化还原催化剂

• 批准号: 8517334
• 负责人: PRAKASH G JAGTAP
• 金额: $ 30.82万
• 依托单位: RADIKAL THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-16 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8517334
• 关键词: 3-nitrotyrosine; Acetylcysteine; Acute; Acute Lung Injury; Address; Animal Model; Area; Attenuated; Biochemical Markers; Blood Circulation; Blood gas; Blood-Air Barrier; Breathing; Bronchoalveolar Lavage; Catalysis; Cell Death; Cells; Cellularity; Chairperson; Complication; Control Groups; DNA Repair Enzymes; Dose; Drug Targeting; Economic Inflation; Environmental air flow; Enzymes; Euthanasia; Experimental Models; F2-Isoprostanes; Female; Free Radicals; Functional disorder; Gases; Gene Expression; Generations; Genetic; Glutathione Disulfide; Heart Rate; Histologic; Hydrogen Peroxide; Infarction; Infiltration; Inflammation; Inflammatory; Inhibitory Concentration 50; Injury; Intensive Care Units; Intercellular adhesion molecule 1; Interruption; Intervention; Intravenous; Isoprostanes; Left; Left lung; Legal patent; Lung; Mechanical ventilation; Mechanics; Medical; Methylprednisolone; Modeling; Monitor; Multiple Organ Failure; Mus; Myocardial Ischemia; Nuclear; Outcome Measure; Oxidants; Oxidation-Reduction; P-Selectin; Pathway interactions; Permeability; Peroxidases; Peroxonitrite; Pharmacodynamics; Pharmacologic Substance; Phase; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerases; Positive-Pressure Respiration; Prevention; Property; Proteins; Pulmonary Edema; Pulse Pressure; Randomized; Rattus; Reperfusion Injury; Resistance; Respiratory Failure; Respiratory System; Resuscitation; Right lung; Risk; Rodent Model; Series; Shapes; Shunt Device; Single-Blind Study; Small Business Innovation Research Grant; Sprague-Dawley Rats; Stress; Sulfhydryl Compounds; TNF gene; Testing; Therapeutic; Tidal Volume; Tissues; Trauma; United States National Institutes of Health; Variant; Ventilator-induced lung injury; Ventricular; Zymosan; base; catalase; catalyst; clinically relevant; hemodynamics; in vivo Model; indexing; inhibitor/antagonist; innovation; lung injury; mimetics; mortality; neutrophil; nitrosative stress; novel; oxidant stress; pressure; prevent; protein expression; public health relevance; pulmonary arterial hypertension; small molecule; treatment effect

DESCRIPTION (provided by applicant): High volume mechanical ventilation (MV) induces a lethal pulmonary complication, termed "ventilator induced lung injury" (VILI), via the generation of toxic free radical and oxidant species that damage the alveolocapillary membrane, trigger pro-inflammatory gene expression, and induce pulmonary arterial hypertension and respiratory failure. To address this unmet medical need, we are developing a novel bifunctional small molecule, R-503, that: 1) prevents the generation of peroxynitrite via broad-spectrum redox catalysis, and 2) competitively inhibits the nuclear DNA repair enzyme poly(ADP-ribose) polymerase 1 ("PARP- 1"). In murine LD100 models of zymosan-induced multiple organ failure (MOF) and Cl2 inhalational lung injury, resuscitation with R-503 begun after the onset of zymosan instillation or Cl2 inhalation reduced neutrophil (PMN) infiltration by 65% and histologic lung injury by 80% (p<0.001). Based upon the centrality of inflammation and redox stress to the pathophysiology of VILI, we hypothesize that PARP-1 is highly activated in the lung and that injury will be attenuated after R-503 intervention. Specific Aim #1: Establish the pharmacodynamic (PD) profile of R-503 in a rat model of VILI Sprague-Dawley rats will be subjected to high tidal volume (TV) MV with positive end expiratory pressure (PEEP) of 3 cm H2O. TV will be initially set at 30 mL/kg and then reduced to limit peak inspiratory pressure (PIP) to 30 cm H2O. Treatment with R-503 (0, 10, 30, and 80 mg/kg) will be initiated 1 h after the onset of high volume MV. A negative control group will undergo low-volume ventilation (TV=6 mL/kg, PEEP=3 cm H2O). A positive control group will undergo high-volume MV plus therapy with IV methylprednisolone (30 mg/kg). A comparator group will be administered IV N-acetylcysteine (NAC) at an equivalent mass dose (30 mg/kg). Lung mechanics will be evaluated hourly, allowing for determination of: 1) dynamic respiratory system compliance, 2) resistance and hysteresivity derived from dynamic pressure volume loops, and 3) static compliance, including lower and upper inflection points derived from the shape of the static inflation and deflation airway pressure curves. We will monitor hemodynamic status by tracking heart rate, mean arterial pressure, and arterial pulse pressure variation. Arterial blood gas determination followed by bronchoalveolar lavage (BALF) of the left lung will be undertaken 4 h after the onset of MV, directly before euthanasia, allowing for determination of gas exchange and the PCO2/Minute Volume Ratio. The BALF will be analyzed for cellularity, LDH release, pro-inflammatory protein expression, and protein content. The right lung will be analyzed for histologic injury, wet/dry ratio, biochemical markers of redox stress (F2¿-isoprostane, GSH/GSSG ratio), PMN infiltration (myeloperoxidase; "MPO"), pro-inflammatory protein expression, and immunohistochemical evidence of peroxynitrite-induced 3- nitrotyrosine (3-NT) and poly(ADP-ribose) formation. Lung concentrations of R-503 will be correlated with outcome measures, so as to construct a PD profile.

描述（由申请人提供）：大容量机械通气（MV）通过产生有毒自由基和氧化物质，损伤肺泡毛细血管膜，引发促炎基因表达，诱发肺动脉高压和呼吸衰竭，诱发致命的肺部并发症，称为“呼吸机诱导肺损伤”（VILI）。为了解决这一未满足的医疗需求，我们正在开发一种新的双功能小分子R-503，它：1)通过广谱氧化还原催化阻止过氧亚硝酸盐的产生，2)竞争性地抑制核DNA修复酶聚（adp -核糖）聚合酶1 （PARP- 1）。在zymosan诱导的多器官衰竭（MOF）和Cl2吸入性肺损伤的小鼠LD100模型中，在开始zymosan灌注或Cl2吸入后开始R-503复苏，使中性粒细胞（PMN）浸润减少65%，组织学肺损伤减少80% （p<0.001）。基于炎症和氧化还原应激在VILI病理生理中的中心地位，我们假设PARP-1在肺中高度激活，R-503干预后损伤将减轻。具体目的#1：建立R-503在VILI大鼠模型中的药效学（PD）谱。将大鼠置于高潮气量（TV） MV下，呼气末正压（PEEP）为3 cm H2O。TV将最初设置为30 mL/kg，然后降低至限制峰值吸气压力（PIP）为30 cm H2O。R-503（0、10、30和80 mg/kg）治疗将在高容量MV开始后1小时开始。阴性对照组进行小容量通气（TV=6 mL/kg， PEEP=3 cm H2O）。阳性对照组将接受大容量MV加静脉注射甲基强的松龙（30mg /kg）治疗。对照组以同等质量剂量（30mg /kg）静脉注射n -乙酰半胱氨酸（NAC）。肺力学将每小时评估一次，以确定：1)动态呼吸系统顺应性，2)阻力和迟滞性来自动态压力容积循环，以及3)静态顺应性，包括由静态充气和充气气道压力曲线形状得出的上下拐点。我们将通过跟踪心率、平均动脉压和动脉脉压变化来监测血液动力学状态。动脉血气测定后，支气管肺泡灌洗（BALF）左肺将在MV发作后4小时，直接在安乐死前进行，允许测定气体交换和二氧化碳分压/分钟体积比。将分析BALF的细胞结构、LDH释放、促炎蛋白表达和蛋白含量。分析右肺组织损伤、湿/干比值、氧化还原应激生化指标（F2¿-异前列腺素，GSH/GSSG比值）、PMN浸润（髓过氧化物酶；“MPO”）、促炎蛋白表达，以及过氧亚硝酸盐诱导的3-硝基酪氨酸（3- nt）和聚adp核糖形成的免疫组织化学证据。肺R-503浓度将与预后指标相关，从而构建PD概况。