Restoration of free radical homeostasis: novel therapy of septic shock

 描述（由申请人提供）：Radikal Therapeutics（RTX）发明了一种新型双功能一氧化氮（NO）供体和氧化还原催化剂（R-190），用于治疗脓毒性休克。在内毒素血症和革兰氏阴性杆菌感染性休克的啮齿动物和绵羊模型中，R-190治疗性复苏改善了临床相关终点的结局，包括血流动力学、氧合和终末器官损伤。在内毒素血症的LD 100鼠模型中，LPS后给予R-100或R-190剂量依赖性地阻断肾、肺和肝损伤73- 90%，抑制肝、肾、肺和肠中的组织学损伤75- 90%，并确保100%存活。在假单胞菌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。拟定的工作范围将在临床相关的脓毒性休克模型中构建PD特征，设计和实施释放和跟踪活性药物成分的方法，并开发R-190及其代谢产物的生物分析方法。目标一：在脓毒症绵羊模型中建立R-190的药效学（PD）特征RTX将进行一项安慰剂对照随机化研究，其中在机械通气雄性美利奴羊的LD 100模型中通过IV途径接种铜绿假单胞菌，产生脓毒性休克。我们将开展研究，以确定R-190在脓毒症绵羊中的剂量反应、时间窗和安全性。任务一：将R-190（30、100和300 mg/kg IV q6 h IV）与媒介物对照进行比较，其中初始剂量在菌血症发作后1小时递送，以建立提供最佳结果的最低剂量（“LDP 00”）。每6小时测量一次R-190的血浆水平，以将药物浓度与疗效联系起来。任务二：我们将通过在菌血症发作后1、2、4、8、12或24 h（LDPOO剂量）开始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、氧合和呼吸参数。在菌血症发作后48小时检查血清和组织，以确定对R-190的反应。目标2：合成R-190。开发放行和稳定性分析方法。确定杂质和降解产物，从而确定最佳储存条件和给药制剂。R-190 API材料在用于绵羊研究前将进行鉴定，以 以确保身份和纯度。为此，将按照ICH指导原则开发分析方法，以确定R-190 API和给药溶液的储存条件和稳定性。目标3：开发定量血浆中R-190及其代谢物的生物分析方法RTX将构建R-190及其代谢物血浆浓度与药理活性相关的药效学特征。将开发一种耐用的生物分析LC-MS/MS方法，其LOQ比现有方法低2 log，允许在低ng/mL水平下鉴别和定量血浆R-190及其主要代谢产物。我们将对该方法的线性、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储存条件。

 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.