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）和超氧阴离子的生物合成的改变通常被认为通过诱导微血管缺血和直接器官毒性而导致败血症中的广泛组织损伤。为了纠正这种自由基失衡，Radikal Therapeutics (RTX) 正在开发一种一流的小分子药物 (R-100)，这是一种基于双功能氧化还原的技术，由 2 个化学部分共价连接而成：1) 一种释放 NO 的有机硝基血管舒张剂，2) 一种吡咯烷氮氧化物，可作为反应活性的三功能催化剂。 氧物质降解：超氧化物歧化酶模拟物、过氧化氢酶模拟物（可解毒过氧化氢）和过氧亚硝酸盐分解催化剂。在内毒素血症的 LD100 小鼠模型中，脂多糖 (LPS) 攻击后 1 小时开始使用 R-100 进行复苏可实现 100% 存活，同时几乎完全防止终末器官损伤。我们假设 R-100 优于其两种成分功能的总和，并且这两种特性共价融合成单个分子实体为脓毒症治疗创造了强大的商业前景。 RTX 将通过进行内毒素研究来测试这一假设，以确定腹膜内 (IP) 施用 R-100 的小鼠的剂量反应、时间窗、作用机制和安全性。目标#1：在脂多糖 (LPS) 攻击后 1 小时，将 3 个剂量水平的 R-100 与载体对照进行比较，以确定提供最佳结果的最低剂量 (“LDPOO”)。将测量 R-100 以及来自心脏、肺、肾和肝脏的代谢物的组织水平，以便将血浆和器官药物摄取联系起来。目标#2：将 R-100 的 LDPOO 剂量与等摩尔剂量的羟甲基丙氧基（“HMP”，R-100 的硝基氧成分）、单硝酸异山梨酯（“ISMN”，经典的单功能 NO 供体）以及 HMP 和 ISMN 的组合进行比较，以验证双功能化合物（R-100）是 优于其组件功能的混合。 LPS 激发后 1 小时开始治疗。目标#3：我们将通过在 LPS 激发后 1、2、4 和 8 小时引入 R-100 来确定治疗时间窗的持续时间。在上述每个目的中，LPS给药后48小时检查血清和组织，以确定形态学和生化终点，包括脂质过氧化（F21-异前列腺素、GSH:GSSG比率）、中性粒细胞浸润、3-硝基酪氨酸（3-NT)、亚硝酸盐/硝酸盐和聚（ADP-核糖）形成， BUN、肌酐、NGAL、AST、ALT、组织学损伤评分以及 TNF-1、HMGB-1、IL-6、TRP14、Trx1、LC8、I:B1、IL-12、MIP-11、CXCL9 和 CXCL10 的血清浓度。目标#4：在 4 小时内，我们将监测麻醉内毒素血症小鼠的外周动脉血压和心率，这些小鼠在 LPS 攻击后 1 小时使用载体对照或 R-100 进行治疗，以确认治疗方案的血流动力学中性。我们预计 R-100 单独或联合使用均优于 HMP 和 ISMN，并且在 LPS 攻击后 6 小时启动时有效，从而验证其作为临床脓毒症潜在治疗剂的实用性。 公共卫生相关性：急性肠穿孔和感染引起的感染性休克是死亡的主要原因。目前，这种情况还没有批准的治疗方法，预后普遍较差。我们正在开发一种针对感染性休克基本机制的新药，并已证明在感染性炎症的实验模型中有效。我们现在将在一系列研究中测试这种药物，以确定最佳剂量，确认作用机制，并确定疾病发作后可以开始治疗的机会窗口。