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）：高容量机械通气（MV）通过产生毒性自由基和氧化剂物质诱导致死性肺部并发症，称为“呼吸机诱导的肺损伤”（VILI），这些物质损伤肺泡毛细血管膜，触发促炎基因表达，并诱导肺动脉高压和呼吸衰竭。为了解决这一未满足的医疗需求，我们正在开发一种新型双功能小分子R-503，其：1）通过广谱氧化还原催化防止过氧亚硝酸盐的生成，2）竞争性抑制核DNA修复酶聚（ADP-核糖）聚合酶1（“PARP- 1”）。在酵母聚糖诱导的多器官衰竭（MOF）和Cl 2吸入性肺损伤的小鼠LD 100模型中，在酵母聚糖滴注或Cl 2吸入开始后用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对VILI病理生理学的中心作用，我们假设PARP-1在肺中高度活化，并且在R-503干预后损伤将减弱。具体目标1：在VILI大鼠模型中建立R-503的药效学（PD）特征Sprague-Dawley大鼠将接受高潮气量（TV）MV和3 cm H2O的呼气末正压（PEEP）。TV最初设定为30 mL/kg，然后降低至吸气峰压（PIP）限制为30 cm H2O。在高容量MV发作后1小时开始R-503（0、10、30和80 mg/kg）给药。阴性对照组将接受低容量通气（TV=6 mL/kg，PEEP=3 cm H2O）。阳性对照组将接受IV甲泼尼龙（30 mg/kg）的高容量MV+治疗。对照组将以等效质量剂量（30 mg/kg）IV给予N-乙酰半胱氨酸（NAC）。将每小时评价肺力学，以确定：1）动态呼吸系统顺应性，2）来自动态压力容积环的阻力和阻力系数，以及3）静态顺应性，包括来自静态充气和放气气道压力曲线形状的上下拐点。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流动力学状态。在MV发作后4小时，即安乐死前即刻进行左肺支气管肺泡灌洗（BALF）后的动脉血气测定，以测定气体交换和PCO 2/分钟容积比。将分析BALF的细胞构成、LDH释放、促炎蛋白表达和蛋白含量。将分析右肺的组织学损伤、湿/干比、氧化还原应激的生化标志物（F2-异前列腺素，GSH/GSSG比）、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。R-503的肺浓度将与结局指标相关，以构建PD特征。