A Redox-active PARP Inhibitor for Lung Transplantation

用于肺移植的氧化还原活性 PARP 抑制剂

基本信息

批准号：
8391286
负责人：
ANDREW Lurie SALZMAN
金额：
$ 39.07万
依托单位：
RADIKAL THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8391286
关键词：
3-nitrotyrosine Acute Acute Lung Injury Animal Model Animals Apoptosis Area Breathing Bronchoalveolar Lavage Catalysis Cells Cellularity Chairperson Chemistry Chlorine Clinical Consumption Cytoprotective Agent DNA Repair Enzymes Dose Drug Delivery Systems European Excision Exhibits Experimental Models F2-Isoprostanes Flushing Genes Genetic Histologic Histology Hydrogen Peroxide IL6 gene Immunohistochemistry In Situ Infarction Inflammation Inflammatory Inhibitory Concentration 50 Injury Intercellular adhesion molecule 1 Interleukin-1 Interleukin-6 Interruption Ischemia Isoprostanes Left Left lung Legal patent Life Lipid Peroxidation Lung Lung Compliance Lung Transplantation MAPK14 gene MAPK8 gene Macrophage Inflammatory Protein-1 Measures Mediating Messenger RNA Mitogen-Activated Protein Kinases Modeling Morbidity - disease rate Multiple Organ Failure Mus Myocardial Ischemia Necrosis Neutrophil Infiltration Nitrosation North Carolina Nuclear Organ Transplantation Orphan Drugs Oxidation-Reduction P-Selectin Pathway interactions Perfadex Permeability Pharmaceutical Preparations Pharmacologic Substance Pharmacotherapy Phosphorylation Pneumonia Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Population Prevention Process Property Proteins Pulmonary Edema Pulmonary Vascular Resistance Rattus Relative (related person)Reperfusion Injury Reperfusion Therapy Residual state Respiratory Failure Resuscitation Reverse Transcriptase Polymerase Chain Reaction Risk Rodent Model Shock Shunt Device Single-Stranded DNA Small Inducible Cytokine A3 Societies Sprague-Dawley Rats Stress Structure of parenchyma of lung Sulfhydryl Compounds TNF gene TNFRSF5 gene Technology Testing Therapeutic Thoracotomy Time Tissues Translations Universities Vascular Permeabilities Weight Zymosan catalase catalyst clinically relevant commercialization drug market exhaust experience in vivo Model indexing inflammatory marker inhibitor/antagonist innovation lung injury lung ischemia medical complication mimetics mortality nitrosative stress novel novel therapeutics oxidant stress prevent professor prophylactic protein B

项目摘要

DESCRIPTION (provided by applicant): Lung ischemia-reperfusion injury (IRI) is an acute lung injury that contributes to morbidity and mortality following lung transplantation (LTX). IRI i mediated in part by the synthesis of oxidative and nitrosative species that in turn induce DNA single strand breakage and triggering of the nuclear DNA repair enzyme, poly(ADP-ribose) polymerase-1 (PARP-1). Hyperactivation of PARP-1 results in the consumption of its substrate NAD, which in turn exhausts intracellular energetics, leading to ATP depletion and tissue necrosis. PARP-1 activation also induces a widespread expression of pro-inflammatory genes that contribute to vascular permeability, lung edema, neutrophil infiltration, pulmonary shunt, and respiratory failure. In models of IRI injury, genetic deletion or pharmacologic inhibition of PARP-1 potently reduces lung damage, but the extent of protection is subtotal. We propose a more thorough abrogation of IRI-induced tissue damage via the deployment of a bifunctional PARP-1 inhibitor, R-503, formed from the covalent linkage of 2 moieties, each with demonstrated tissue protection: 1) a PARP-1 inhibitor moiety, and 2) a thiol-rich dihydrolipoyl ("DHL") domain that acts as a broad-spectrum redox degradation catalyst. Aim #1: Establish the superiority of the bifunctional PARP-1 inhibitor, R-503, relative to its component domains, in a rat model of warm-ischemic lung IRI. Rat lungs are rendered ischemic in situ for 60 min and reperfused for 4 h. Prior to ischemia, rats are treated with IV R-503, DHL, the monofunctional PARP-1 inhibitor INO-1001, or a combination of DHL and INO-1001. Tissue damage is assessed by histology, levels of PMN infiltration, lipid peroxidation, NF-?B, protein nitrosation, PARP-1 activation, apoptosis, and oxygenation (PaO2). Inflammation is assessed by examining BAL markers of inflammation (protein, PMNs, TNF-¿, and MIP-1¿). Aim #2: Establish the efficacy of R-503 in a syngeneic rat model of orthotopic LTX. Using a translational model of LTX, donor rats are treated IV with R-503 or vehicle 10 min before lung removal. After flushing with cold Perfadex", spiked with R-503 (30 ¿M) or vehicle, donor lungs are stored cold for 12 h before left LTX. Right donor lungs are processed for immunohistochemical analysis of PARP-1 activation. Immediately following LTX with left donor lungs, recipients will receive R-503 or vehicle. The dose of R-503 will be guided by the optimal level elucidated in Aim #1. Recipient rats will be evaluated at 3 different reperfusion times (1, 3, 6 h) for wet/dry weight ratio, graft oxygenation, pulmonary vascular resistance, dynamic compliance, and lung F2¿-isoprostane, histology, and immunohistochemistry for 3-nitrotyrosine and poly(ADP-ribose). Specific analyses will be carried out at 3 time points: at 1 h post reperfusion for I?B¿, phosphorylation of MAP kinases, nuclear p50; at 3 h post reperfusion for RT-PCR to quantify lung 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¿, IL6, and IL1-¿. PUBLIC HEALTH RELEVANCE: Ischemia-reperfusion injury is a major medical complication following lung transplantation and contributes to the high mortality in this population. At present there are no approved prophylactic measures. We are developing a novel drug that targets the basic mechanisms of this condition and will test this agent in two clinically-relevant animal models.

描述（由申请人提供）：肺缺血再灌注损伤（IRI）是一种急性肺损伤，导致肺移植（LTX）后的发病率和死亡率，部分由氧化和亚硝化物质的合成介导，而氧化和亚硝化物质的合成又反过来诱导。 DNA 单链断裂并触发核 DNA 修复酶、聚（ADP-核糖）聚合酶-1 (PARP-1) 的过度激活。 PARP-1 导致其底物 NAD 的消耗，进而耗尽细胞内能量，导致 ATP 耗竭和组织坏死。 PARP-1 激活还会诱导促炎基因的广泛表达，从而导致血管通透性、肺水肿、在 IRI 损伤模型中，PARP-1 的基因缺失或药物抑制会导致中性粒细胞浸润、肺分流和呼吸衰竭，但保护程度较低。我们建议通过部署双功能 PARP-1 抑制剂 R-503 更彻底地消除 IRI 引起的组织损伤，R-503 由 2 个部分共价连接而成，每个部分都具有组织保护作用：1) PARP-1抑制剂部分，以及 2) 富含硫醇的二氢硫辛酰 (“DHL”) 结构域，充当广谱氧化还原降解催化剂。目标 1：确立该结构的优越性。双功能 PARP-1 抑制剂 R-503，在热缺血性肺 IRI 大鼠模型中，使大鼠肺原位缺血 60 分钟，然后再灌注 4 小时。使用 IV R-503、DHL、单功能 PARP-1 抑制剂 INO-1001 或 DHL 和 INO-1001 的组合来评估组织损伤。组织学、PMN 浸润水平、脂质过氧化、NF-κB、蛋白质亚硝化、PARP-1 激活、细胞凋亡和氧合 (PaO2) 通过检查 BAL 炎症标志物（蛋白质、PMN、TNF-¿）来评估炎症。，和MIP-1¿目标#2：建立 R-503 在原位 LTX 大鼠模型中的功效，使用 LTX 转化模型，在肺切除后 10 分钟用 R-503 或载体进行静脉注射。 Perfadex”，掺有 R-503 (30 µM) 或媒介物，供体肺在左 LTX 之前冷藏 12 小时。右供体肺经过处理对左供体肺进行 LTX 后立即进行 PARP-1 激活的免疫组织化学分析。R-503 的剂量将根据目标 #1 中阐明的最佳水平进行评估。湿/干重比、移植物的 3 种不同再灌注时间（1、3、6 小时）氧合、肺血管阻力、动态顺应性和肺 F2¿ - 3-硝基酪氨酸和聚（ADP-核糖）的异前列腺素、组织学和免疫组织化学将在 3 个时间点进行具体分析：再灌注后 1 小时进行 I?B¿ , 磷酸化 MAP 激酶、核 p50；再灌注 3 小时后进行 RT-PCR，以定量 TNF-¿ ，MIP-1??和 Bcl-2；并在再灌注后 6 小时测定 BALF 细胞结构、蛋白质浓度、TNF-¿ ，MIP-1?? 、IL6 和 IL1-¿ 。公共卫生相关性：缺血再灌注损伤是肺移植后的主要并发症，导致该人群的高死亡率。目前，我们正在开发一种针对这种情况的基本机制的新药物。将在两种临床相关的动物模型中测试该药物。