Restoration of free radical homeostasis: novel therapy of septic shock

恢复自由基稳态：感染性休克的新疗法

• 批准号: 9140177
• 负责人: ANDREW Lurie SALZMAN
• 金额: $ 124.86万
• 依托单位: RADIKAL THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9140177
• 关键词: 3-nitrotyrosine; Anabolism; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Bacteremia; Biochemical; Biological; Biological Markers; Blood flow; Choking; Clinical Research; Clinical Trials; Collaborations; Conscious; Creatinine; Creatinine clearance measurement; Critical Illness; Diagnosis; Dose; Endotoxemia; Epithelial; Evaluation; Excision; Excretory function; Formulation; Free Radicals; Functional disorder; Glucose; Glutathione Disulfide; Guidelines; Health; High Pressure Liquid Chromatography; Histologic; Homeostasis; Infiltration; Inflammatory; Injury; Isoprostanes; Kidney; LCN2 gene; Liver; Lung; Measurement; Measures; Mediating; Medical; Methodology; Methods; Modeling; Multi-Institutional Clinical Trial; Muscle Tonus; Nitric Oxide; Nitric Oxide Donors; Nitrogen; Organ; Outcome; Outcome Measure; Outcome Study; Oxidation-Reduction; Oxygen; Patients; Peritonitis; Peroxonitrite; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Phase; Phase I Clinical Trials; Plasma; Preparation; Procedures; Prodrugs; Product R; Protocols documentation; Pseudomonas aeruginosa; Qualifying; Rattus; Reaction Time; Reference Standards; Reperfusion Injury; Resuscitation; Rodent; Route; Safety; Sepsis; Septic Shock; Serum; Sheep; Side; Sodium; Solid; Superoxides; Testing; Texas; Therapeutic; Therapeutic Agents; Tight Junctions; Time; Tissues; Toxicology; Universities; Vascular Smooth Muscle; Water; Work; analytical method; catalyst; chemokine; clinically relevant; cytokine; extracellular; hemodynamics; immunoreactivity; improved; improved outcome; liver injury; male; model design; mouse model; novel; novel therapeutics; preclinical trial; pulmonary arterial hypertension; randomized placebo controlled trial; renal ischemia; response; restoration; septic

 DESCRIPTION (provided by applicant): Radikal Therapeutics (RTX) has invented a novel first-in-class bifunctional nitric oxide (NO) donor and redox catalyst (R-190) to treat septic shoc. In rodent and ovine models of endotoxemia and gram negative bacillary septic shock, therapeutic resuscitation with R-190 improves outcome across clinically-relevant endpoints, including hemodynamics, oxygenation, and end-organ injury. In an LD100 murine model of endotoxemia, post-LPS administration of R-100 or R-190 dose-dependently blocked renal, lung, and hepatic injury 73-90%, inhibited histologic damage in liver, kidney, lung, and gut by 75-90%, and assured 100% survival. In an ovine model of Pseudomonal septic shock, R-190 resuscitation restored hemodynamics and oxygenation. The proposed scope of work will construct a PD profile in a clinically-relevant septic shock model, design and implement methods to release and track the active pharmaceutical ingredient, and develop bioanalytical methods of R-190 and its metabolites. Aim #1: Establish the pharmacodynamic (PD) profile of R-190 in an ovine model of sepsis RTX will carry out a placebo-controlled randomized study wherein septic shock is produced in an LD100 model in mechanically-ventilated male Merino sheep administered an inoculum of Pseudomonas aeruginosa via an IV route. We will carry out studies in order to establish the dose-response, time-window, and safety of R-190 in the septic sheep. Task #1: R-190 (30, 100, and 300 mg/kg IV q6h IV) will be compared to vehicle control wherein the initial dose is delivered 1 h post onset of bacteremia, in order to establish the lowes dose providing optimal outcome ("LDPOO"). Plasma levels of R-190 will be measured q6h, to relate drug concentration to efficacy. Task #2: We will determine the duration of the therapeutic time window by initiating R-190 therapy 1, 2, 4, 8, 12, or 24 h (at the LDPOO dose) after the onset of bacteremia. Hemodynamic, oxygenation, and ventilatory parameters will be assessed q3h. Serum and tissue will be examined 48 h post onset of bacteremia for determination of response to R-190. Aim #2: Synthesize R-190. Develop analytical methods for release and stability. Define impurities and degradants and thereby define optimal storage conditions and dosing formulation. R-190 API material will be qualified prior to use in the ovine studies in order to assure identity and purity. Analytical methods will be developed following ICH guidelines for this purpose and to establish storage conditions and stability of the R-190 API and dosing solutions. Aim #3: Develop a bioanalytical methodology to quantitate R-190 and its metabolites in plasma RTX will construct a pharmacodynamic profile relating plasma concentrations of R-190 and its metabolites to pharmacologic activity. A robust bioanalytical LC-MS/MS approach will be developed with an LOQ 2 logs less than existing methods, allowing for identification and quantitation of plasma R-190 and its major metabolites at the low ng/mL level. We will qualify this approach for linearity, precision, and accuracy, and define the conditions of plasma preparation and storage that optimize reliability of this methodology.

 描述(由申请人提供)：Radikal Treateutics(RTX)发明了一种新的一流的双功能一氧化氮(NO)供体和氧化还原催化剂(R-190)，用于治疗败血症。在内毒素血症和革兰氏阴性杆菌败血症休克的啮齿动物和绵羊模型中，R-190的治疗性复苏改善了临床相关终点的预后，包括血流动力学、氧合和终末器官损伤。在LD100小鼠内毒素血症模型中，LPS后给予R-100或R-190可剂量依赖性地阻断肾、肺和肝损伤73-90%，抑制肝、肾、肺和肠道的组织学损伤75%-90%，并确保100%的存活率。在假性感染性休克的绵羊模型中，R-190复苏恢复了血流动力学和氧合。拟议的工作范围将在临床相关的感染性休克模型中构建PD概况，设计和实施释放和跟踪活性药物成分的方法，并开发R-190及其代谢物的生物分析方法。目的#1：建立R-190在绵羊脓毒症模型中的药效学(PD)模型RTX将进行一项安慰剂对照随机研究，在LD100模型中，通过静脉注射铜绿假单胞菌，在机械通气的雄性美利奴绵羊中产生感染性休克。我们将开展研究，以确定R-190在败血症绵羊身上的剂量反应、时间窗口和安全性。任务1：将R-190(30、100和300 mg/kg静脉注射，q6h静脉注射)与赋形剂对照进行比较，其中初始剂量在菌血症发生后1小时给予，以确定提供最佳结果的低剂量(“LDPOO”)。将每隔6小时测量一次血浆R-190水平，以将药物浓度与疗效联系起来。任务2：我们将通过在菌血症发生后1、2、4、8、12或24小时(按LDPOO剂量)启动R-190治疗来确定治疗时间窗的持续时间。每3小时评估一次血流动力学、氧合和呼吸机参数。血清和组织将在菌血症发病48小时后进行检查，以确定对R-190的反应。目标2：合成R-190。开发释放和稳定性的分析方法。定义杂质和降解剂，从而确定最佳储存条件和配料配方。R-190原料药材料将在用于绵羊研究之前进行鉴定，以便 以确保身份和纯洁。为此目的，将按照ICH指南开发分析方法，并确定R-190原料药和配料溶液的储存条件和稳定性。目的#3：建立R-190及其代谢物在血浆中定量的生物分析方法RTX将建立R-190及其代谢物血药浓度与药理活性之间的药效学曲线。将开发一种可靠的生物分析LC-MS/MS方法，其LOQ 2log小于现有方法，从而能够在低ng/mL水平上鉴定和定量血浆R-190及其主要代谢物。我们将对该方法的线性度、精密度和准确度进行鉴定，并确定优化该方法可靠性的血浆制备和存储条件。