Peroxynitrite Decomposition Catalyst and Nitric Oxide Donor for Endotoxemia

过氧亚硝酸盐分解催化剂和一氧化氮供体治疗内毒素血症

• 批准号: 8248638
• 负责人: Garry John Southan
• 金额: $ 25.23万
• 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 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8248638
• 关键词: 3-nitrotyrosine; Acute; Anabolism; Arginine; Biochemical; Biological; Blood Pressure; CXCL10 gene; CXCL9 gene; Chemicals; Clinical; Complex; Consumption; Creatinine; Dose; Drug Delivery Systems; Endotoxemia; Endotoxic Shock; Enzymes; European; Event; Excision; Experimental Models; F2-Isoprostanes; Free Radicals; Functional disorder; Glucose; Glutathione Disulfide; Grant; HMGB Proteins; Heart; Heart Rate; Histology; Hydrogen Peroxide; Infection; Infiltration; Inflammation; Injury; Interleukin-12; Interleukin-6; Intestines; Investigation; Ischemia; Isoprostanes; Kidney; Lipid Peroxidation; Lipopolysaccharides; Liver; Lung; Macrophage Inflammatory Protein-1; Measures; Microcirculation; Mitochondria; Modeling; Molecular; Monitor; Multiple Organ Failure; Mus; NADPH Oxidase; Neutrophil Infiltration; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitrites; Nitrogen; Onset of illness; Organ; Outcome; Oxidation-Reduction; Oxygen; Perforation; Peripheral; Peroxonitrite; Pharmaceutical Preparations; Pharmacodynamics; Plasma; Poly Adenosine Diphosphate Ribose; Production; Property; Protein Isoforms; Pyrrolidines; Rattus; Reaction; Reaction Time; Reactive Oxygen Species; Reperfusion Injury; Resuscitation; Rodent Model; Safety; Secondary to; Sepsis; Septic Shock; Series; Serum; Shock; Societies; Sum; Superoxide Dismutase; Superoxides; TNF gene; Technology; Testing; Therapeutic; Time; Tissues; Toxic effect; Treatment Protocols; Validation; Water; Xanthine Oxidase; base; catalase; catalyst; hemodynamics; human NOS2A protein; human NOS3 protein; innovation; intraperitoneal; male; meetings; mimetics; mortality; neutrophil; novel; outcome forecast; professor; pyrrolidine; renal ischemia; response; septic; small molecule; tetrahydrobiopterin; therapeutic target; uptake

DESCRIPTION (provided by applicant): Alterations in the biosynthesis of the free radicals nitric oxide (NO) and superoxide anion are generally accepted to contribute to widespread tissue injury in sepsis via their induction of microvacular ischemia and direct organ toxicity. To correct this free radical imbalance, Radikal Therapeutics (RTX) is developing a first- in-class small molecule drug (R-100), a bifunctional redox-based technology formed from the covalent linkage of 2 chemical moieties: 1) an organic nitrovasodilator that releases NO, and 2) a pyrrolidine nitroxide that acts as a trifunctional catalyst of reactive oxygen species degradation: a superoxide dismutase mimetic, a catalase mimic that detoxifies hydrogen peroxide, and a peroxynitrite decomposition catalyst. In an LD100 murine model of endotoxinemia, resuscitation by R-100 starting 1 h after lipopolysaccharide (LPS) challenge produces 100% survival, accompanied by near complete protection against end-organ injury. We hypothesize that R-100 is superior to the sum of its two component functionalities and that the covalent fusion of these two properties into a single molecular entity creates a strong commercial prospect for therapy of sepsis. RTX will test this hypothesis by carrying out endotoxemic studies to establish the dose-response, time-window, mechanism of action, and safety in mice of intraperitoneal (IP) administered R-100. Aim #1: R-100 at 3 dose levels will be compared to vehicle control 1 h post lipopolysaccharide (LPS) challenge, in order to establish the lowest dose providing optimal outcome ("LDPOO"). Tissue levels of R-100 and metabolites from heart, lung, kidney, and liver will be measured, in order to relate plasma and organ drug uptake. Aim #2: the LDPOO dose of R-100 will be compared to equimolar doses of hydroxymethylproxyl ("HMP", the nitroxide component of R-100), isososorbide mononitrate ("ISMN", a classic monofunctional NO donor), and the combination of HMP and ISMN, in order to verify that a bifunctional compound (R-100) is superior to a mixture of its component functionalities. Treatment will be initiated 1 h after LPS challenge. Aim #3: We will determine the duration of the therapeutic time window by introducing R-100 1, 2, 4, and 8 h after LPS challenge. Serum and tissue will be examined 48 h post LPS dosing in each of the above Aims for determination of morphologic and biochemical endpoints, including lipid peroxidation (F21-isoprostane, GSH:GSSG ratio), neutrophil infiltration, 3- nitrotyrosine (3-NT), nitrite/nitrate, and poly(ADP-ribose) formation, BUN, creatinine, NGAL, AST, ALT, histology injury score, and serum concentrations of TNF-1, HMGB-1, IL-6, TRP14, Trx1, LC8, I:B1, IL-12, MIP-11, CXCL9, and CXCL10. Aim #4: Over a 4 h period, we will monitor peripheral arterial blood pressure and heart rate in anesthetized endotoxinemic mice treated with vehicle control or R-100 administered 1 h after LPS challenge, in order to confirm the hemodynamic neutrality of the treatment regimen. We expect that R- 100 will be superior to HMP and ISMN, alone and in combination, and will be effective when initiated 6 h after LPS challenge, thereby verifying its utility as a potential therapeutic for clinical sepsis. PUBLIC HEALTH RELEVANCE: Septic shock resulting from acute bowel perforation and infection is a major cause of mortality. At present, there is no approved therapy for this condition and prognosis is uniformly poor. We are developing a novel drug that targets the basic mechanisms of septic shock, and has proven effective in experimental models of septic inflammation. We will now test this agent in a series of investigations in order to determine the optimal dose, to confirm the mechanism of action, and to establish the window of opportunity after disease onset within which therapy may be initiated.

描述（由申请人提供）：自由基一氧化氮（NO）和超氧化阴离子的生物合成的改变通常被接受，可以通过诱导微交流局部缺血和直接器官毒性来促进败血症的广泛组织损伤。 To correct this free radical imbalance, Radikal Therapeutics (RTX) is developing a first- in-class small molecule drug (R-100), a bifunctional redox-based technology formed from the covalent linkage of 2 chemical moieties: 1) an organic nitrovasodilator that releases NO, and 2) a pyrrolidine nitroxide that acts as a trifunctional catalyst of reactive氧降解：一种超氧化物歧化酶模拟酶，一种过氧化氢酶模拟于过氧化氢排毒，并且过氧亚硝酸盐分解催化剂。在内毒素血症的LD100鼠模型中，脂多糖（LPS）挑战后1小时从R-100复苏产生100％的生存率，并伴随着几乎完全完全的保护，以抵抗最终器官损伤。我们假设R-100优于其两个成分功能的总和，并且这两种特性与单个分子实体的共价融合为败血症治疗提供了强大的商业前景。 RTX将通过进行内毒素学研究来检验这一假设，以建立腹膜内（IP）施用R-100的小鼠的剂量反应，时间赋，作用机理和安全性。 AIM＃1：将3剂量水平的R-100与脂多糖后1小时（LPS）挑战的车辆对照进行比较，以确定最低剂量提供最佳结果（“ LDPOO”）。将测量来自心脏，肺，肾脏和肝脏的R-100和代谢物的组织水平，以相关血浆和器官药物的吸收。 Aim #2: the LDPOO dose of R-100 will be compared to equimolar doses of hydroxymethylproxyl ("HMP", the nitroxide component of R-100), isososorbide mononitrate ("ISMN", a classic monofunctional NO donor), and the combination of HMP and ISMN, in order to verify that a bifunctional compound (R-100) is优于其组件功能的混合物。 LPS挑战后1小时将开始治疗。 AIM＃3：我们将通过引入LPS挑战后的R-100 1、2、4和8小时来确定治疗时间窗口的持续时间。 LPS剂量的48小时，以上每个旨在确定形态学和生化终点，包括脂质过氧化（F21-异丙烷，GSH：GSSG：GSSG：GSH：GSSG比率），中性粒细胞浸润，3-硝基含量（3-NT），Nitrate and Nitrate and-nitrate，Nitrate（3-硝酸盐），将检查血清和组织。 BUN，肌酐，Ngal，AST，ALT，组织学损伤评分和TNF-1，HMGB-1，IL-6，TRP14，TRP14，TRX1，LC8，I：B1，IL-11，MIP-11，MIP-11，CXCL9和CXCL10的血清浓度。 AIM＃4：在4小时内，我们将监测用媒介物对照治疗的麻醉性内毒素小鼠外周动脉血压和心率，或在LPS挑战后1小时治疗，以确认治疗方案的血液动力学中度。我们预计R-100单独和组合将优于HMP和ISMN，并且在LPS挑战后6小时开始时将有效，从而验证其效用是对临床败血症的潜在治疗方法。 公共卫生相关性：急性肠穿孔和感染引起的败血性休克是死亡率的主要原因。目前，对于这种情况尚无批准的治疗，预后均匀差。我们正在开发一种针对败血性休克的基本机制的新型药物，并已证明在化粪池炎症的实验模型中有效。现在，我们将在一系列研究中测试该药物，以确定最佳剂量，确认作用机理，并在疾病发作后建立机会窗口，在该疾病开始时可以开始治疗。