Hybrid Katp Channel Opener and Redox Catalyst for Lung Transplantation

用于肺移植的混合 Katp 通道开放剂和氧化还原催化剂

• 批准号: 8451621
• 负责人: PRAKASH G JAGTAP
• 金额: $ 29.63万
• 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: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8451621
• 关键词: 3-nitrotyrosine; Accounting; Acetylcysteine; Action Potentials; Adverse effects; Animal Model; Animals; Antioxidants; Apoptosis; Arrhythmia; Ascorbic Acid; BCL2 gene; Cardiac; Cellularity; Clinic; Clinical; Complex; Cytoprotective Agent; Diazoxide; Dose; Drug Targeting; European; Excision; Exhibits; F2-Isoprostanes; Flushing; Functional disorder; Histologic; Histology; Hybrids; Hydrogen Peroxide; Hydroxyl Radical; Hyperglycemia; Hypotension; In Situ; Infarction; Infiltration; Inflammation; Injury; Interleukin-1; Interleukin-6; Ischemia; Ischemic Preconditioning; Isoprostanes; Kinetics; Left; Left lung; Lipid Peroxidation; Lung; Lung Transplantation; MAPK14 gene; MAPK8 gene; Macrophage Inflammatory Protein-1; Measures; Messenger RNA; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; NF-kappa B; Nitrosation; North Carolina; Nuclear; Organ; Oxidation-Reduction; Pathogenesis; Pathologic; Pathway interactions; Perfadex; Peroxonitrite; Pharmacotherapy; Phosphorylation; Physiological; Pinacidil; Poly Adenosine Diphosphate Ribose; Population; Prevention; Property; Protein Isoforms; Proteins; Pulmonary Edema; Pulmonary Vascular Resistance; Pyrrolidines; Rattus; Reaction; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Reverse Transcriptase Polymerase Chain Reaction; Rodent Model; Route; Secondary to; Shock; Signal Transduction; Small Inducible Cytokine A3; Societies; Sprague-Dawley Rats; Stress; Structure of parenchyma of lung; Superoxide Dismutase; Superoxides; TNF gene; TNFRSF5 gene; Testing; Therapeutic; Thoracotomy; Time; Tissues; Tocopherols; Translating; Transplantation; Universities; Ventricular Fibrillation; Weight; antioxidant therapy; catalase; catalyst; clinically relevant; in vivo; lung injury; lung ischemia; medical complication; mimetics; mortality; novel; preconditioning; professor; prophylactic; public health relevance; pyrrolidine; small molecule; theories; uptake

DESCRIPTION (provided by applicant): Radikal Therapeutics (RTX) is developing a novel cytoprotective agent (R-801) for the prevention of ischemia-reperfusion injury (IRI) following lung transplantation (LTX). In rodent models of severe redox injury, R-801 profoundly reduces organ dysfunction, tissue infarction, and parenchymal inflammation. R- 801 is formed from the covalent fusion of 2 distinct moieties, each with demonstrated tissue protective properties: 1) a mito-K+-ATP channel activating moiety derived from pinacidil, and 2) a pyrrolidine nitroxide domain that acts as superoxide dismutase and catalase mimetics and a peroxynitrite decomposition catalyst. In a murine model of lethal Cl2 inhalational lung injury, R-801 given 2 h after Cl2 exposure blocked all histologic damage (p<0.01), reduced the elevation in nuclear NF-kB by 76% (p<0.0001), and restored the level of IkB¿ to supranormal (p<10-7). Aim #1: Establish the superiority of R- 801 relative to its component functional domains in a rat model of warm-ischemic lung IRI. Left lungs of Sprague Dawley (SD) rats are rendered ischemic in situ for 60 min and reperfused for 4 h. Prior to ischemia, rats are treated with IV R-801 (0, 3, 10, 30 mg/kg), hydroxymethylproxyl (HMP; 30 mg/kg), pinacidil (30 mg/kg), or a combination of HMP and pinacidil (each 30 mg/kg). A sham rat undergoes thoracotomy but neither ischemia nor drug therapy. R-801 is expected to exhibit superior efficacy, relative to treatment with pinacidil, HMP, and their combination, with respect to tissue damage and inflammation. Tissue damage is assessed by examining histologic score, PMN infiltration, lipid peroxidation, protein nitrosation, PARP-1 activation, nuclear NF-kB, apoptosis, and oxygenation (PaO2). Inflammation is assessed by examining BAL for protein, PMNs, TNF-¿, and MIP-1¿. Aim #2: Establish the efficacy of R- 801 in a syngeneic rat model of orthotopic LTX. SD donor rats are treated with R-801 or vehicle control 10 min before lung removal. After flushing with cold Perfadex" spiked with R-801 or vehicle, donor lungs are stored cold for 12h before left LTX. Immediately following LTX with left donor lungs, recipients will receive R-801. Recipient rats will be evaluated for a) wet/dy weight ratio (W/D) (a measure of pulmonary edema), b) oxygenation by the graft, c) graft pulmonary vascular resistance, d) dynamic compliance, and e) lung tissue analysis for F2¿-isoprostane (a measure of lipid peroxidation), histology, and immunohistochemical reactivity to 3-nitrotyrosine (3-NT) and poly(ADP-ribose). Specific analyses will be carried out at 3 time points: at 1 h post reperfusion for IkB¿, nuclear p50, and phosphorylation of mitogen activated protein kinases (MAPKs - ERK, p38, JNK); at 3h post reperfusion for RT-PCR to quantify lung tissue mRNA concentrations of TNF-¿, MIP-1¿, and Bcl-2; and at 6 h post reperfusion for determination of BALF cellularity, protein concentration, TNF-¿, MIP-1¿, IL-6, and IL1-¿. R-801 therapy is expected to translate into decreased primary graft dysfunction and mortality after lung transplant.

描述（由适用提供）：Radikal Therapeutics（RTX）正在开发一种新型的细胞保护剂（R-801），用于预防肺移植后预防缺血 - 再灌注损伤（IRI）（LTX）。在严重氧化还原损伤的啮齿动物模型中，R-801可深刻地减少器官功能障碍，组织违规和副群体感染。 R-801由2个不同的部分的共价融合形成，每个部分具有组织保护特性：1）源自Pinacidil的Mito-K+-ATP通道激活部分，以及2）吡咯烷硝基氧结构域，可作为超氧化物歧化酶和催化酶Mimimimitics和catalase mimimetrits and pecountics nitrits cotostient and cotositient。在致命的CL2吸入肺损伤的鼠模型中，CL2暴露后2小时给予R-801阻断了所有组织学损伤（P <0.01），将核NF-KB的升高降低了76％（P <0.0001），并恢复了IKB的水平至Supranmoral（P <10-7-7）。 AIM＃1：在温暖的缺血性肺IRI大鼠模型中，建立R-801相对于其成分功能结构域的超级性。 Sprague Dawley（SD）大鼠的左肺原位是缺血，持续60分钟，并进行重新注射4小时。在缺血之前，用IV R-801（0、3、10、30 mg/kg），羟基甲基羟基（HMP; 30 mg/kg），Pinacidil（30 mg/kg）或HMP和Pinacidil（每个30 mg/kg）的组合治疗大鼠。假大鼠发生胸腔切开术，但既没有缺血也不是药物治疗。相对于用pinacidil，HMP， 以及它们的结合，就组织损伤和炎症而言。通过检查组织学评分，PMN浸润，脂质过氧化，蛋白质硝化，PARP-1激活，核NF-KB，凋亡和氧合来评估组织损伤（PAO2）。通过检查蛋白质，PMN，TNF-€和MIP-1¿的BAL来评估炎症。 AIM＃2：在原位LTX的合成大鼠模型中建立R-801的效率。 SD供体大鼠在去除肺前10分钟用R-801或车辆对照处理。 After flushing with cold Perfadex" spiked with R-801 or vehicle, donor lungs are stored cold for 12h before left LTX. Immediately following LTX with left donor lungs, recipients will receive R-801. Recipient rats will be evaluated for a) wet/dy weight ratio (W/D) (a measurement of pulmonary edema), b) oxygenation by the graft, c) graft pulmonary血管抗性，D）动态依从性和E）肺组织分析（用于脂质过氧化的测量），组织学和免疫组织化学对3-硝基苯胺（3-NT）的反应性（3-NT）和PORY PORTOR PORTIS PORTIS PORTERS：AN codpose coled and Repb。 RT-PCR的3H再灌注以定量TNF-PCR的肺组织mRNA浓度，MIP-1，MIP-1，和BCl-2的磷酸化（MAPK-ERK，P38，JNK）； IL1- - R-801治疗将转化为肺移植后原发性移植功能障碍和死亡率的降低。