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（由申请人提供）：Radikal Therapeutics (RTX) 正在开发一种新型细胞保护剂 (R-801)，用于预防肺移植 (LTX) 后的缺血再灌注损伤 (IRI)。在严重氧化还原损伤的啮齿动物模型中，R-801 可显着减少器官功能障碍、组织梗塞和实质炎症。 R-801 由 2 个不同部分共价融合而成，每个部分都具有组织保护特性：1) 源自吡那地尔的 mito-K+-ATP 通道激活部分，2) 吡咯烷氮氧结构域，充当超氧化物歧化酶和过氧化氢酶模拟物以及过氧亚硝酸盐分解催化剂。在致命性 Cl2 吸入性肺损伤的小鼠模型中，Cl2 暴露 2 小时后给予 R-801 可阻断所有组织学损伤 (p<0.01)，使核 NF-kB 升高降低 76% (p<0.0001)，并将 IkB¿ 水平恢复至超正常水平 (p<10-7)。目标#1：在热缺血性肺 IRI 大鼠模型中确定 R-801 相对于其组成功能域的优越性。 Sprague Dawley (SD) 大鼠的左肺原位缺血 60 分钟，然后再灌注 4 小时。在缺血之前，用IV R-801（0、3、10、30 mg/kg）、羟甲基丙酰（HMP；30 mg/kg）、吡那地尔（30 mg/kg）或HMP和吡那地尔的组合（各30 mg/kg）治疗大鼠。假大鼠接受开胸手术，但既不进行缺血也不进行药物治疗。相对于吡那地尔、HMP、 以及它们的组合，关于组织损伤和炎症。通过检查组织学评分、PMN 浸润、脂质过氧化、蛋白质亚硝化、PARP-1 激活、核 NF-kB、细胞凋亡和氧合 (PaO2) 来评估组织损伤。通过检查 BAL 中的蛋白质、PMN、TNF-¿ 和 MIP-1¿ 来评估炎症。目标#2：确定 R-801 在原位 LTX 同基因大鼠模型中的功效。 SD 供体大鼠在肺切除前 10 分钟用 R-801 或媒介物对照进行处理。用掺有 R-801 或赋形剂的冷 Perfadex" 冲洗后，供体肺在进行 LTX 之前冷藏 12 小时。在对左供体肺进行 LTX 后，受体将立即接受 R-801。将对受体大鼠进行以下评估：a) 湿重/干重比 (W/D)（肺水肿的衡量标准），b) 移植物的氧合，c) 移植物肺血管 阻力，d) 动态顺应性，e) 肺组织分析 F2¿-异前列烷（脂质过氧化的一种测量方法）、组织学以及对 3-硝基酪氨酸 (3-NT) 和聚 (ADP-核糖) 的免疫组织化学反应性。将在 3 个时间点进行具体分析：再灌注后 1 小时进行 IkB¿、核 p50 和磷酸化 丝裂原激活蛋白激酶（MAPK - ERK、p38、JNK）；再灌注后 3 小时进行 RT-PCR，以定量肺组织 TNF-K、MIP-1K 和 Bcl-2 的 mRNA 浓度；再灌注后 6 小时测定 BALF 细胞结构、蛋白质浓度、TNF-¿、MIP-1Ф、IL-6 和 IL1-Ф。 R-801 疗法有望转化为肺移植后原发性移植物功能障碍和死亡率的降低。