Young, Lindon H

项目摘要 我们正在寻求资金来测试一种新型蛋白激酶C ε（PKCε）的心脏保护作用 抑制剂（YT-002）和一种新的蛋白激酶C β II（PKCβII）抑制剂（YT-004）在猪体内的作用 在再灌注开始时给药时，体内心肌缺血再灌注（I/R）。目前没有 批准的药物治疗/预防I/R损伤。我们充分意识到过去在这方面的失败， 区从字面上看，所有以前的尝试都采取了氧自由基清除方法。我们相信这 这至少部分地解释了为什么没有成功的心脏病药物治疗方法。 I/R损伤至今。一旦产生活性氧，损害是立即的，清除是立即的。 无效。我们的方法-以前从未尝试过-是抑制活性氧的产生， 防止它们在再灌注过程中受损。我们对22名介入心脏病专家进行了访谈 绝大多数人支持这样的观点，即心肌组织和功能的保护将代表一种 对经历缺血-再灌注损伤的患者具有显著的长期益处，导致 心力衰竭的发生率。来自我们实验室的另一种化合物YT-001的先前数据（与化合物YT-001的作用机制相同）。 作用与YT-002相同）在体内表现出对过氧化氢生成的显著抑制， 在离体大鼠I/R心脏中， 在体猪心肌I/R模型。然而，YT-001具有有限的溶解度和剩余专利寿命。我们 新分子（YT-002和YT-004）具有显著改善的溶解性、效力和新的专利寿命。我们 已经在离体大鼠I/R中证明了令人印象深刻的梗死面积减小和心脏功能保护 YT-002和YT-004的心脏。在本建议中，我们将评估YT-002的心脏保护作用， YT-004在Gorman猪模型中对缺血再灌注的作用（19）。除了动物的生命体征， 还将监测分数（通过超声心动图测量）（作为心脏功能/收缩性的指标） 作为心脏蛋白（例如，肌钙蛋白I）作为整个方案中组织损伤的指标。三小时后 在再灌注后，将心脏切除，并染色以直接评估心脏损伤（梗塞大小）。 将在PCOM的Young博士实验室对冷冻心脏切片进行免疫组织化学检测PKCε 和PKCβII定位。基于先前用PKCε抑制剂YT-001产生的猪心数据，我们 预期与接受心脏移植的动物相比， 乱序肽对照。这些积极的数据将为进行更长期的研究提供额外的依据。 动物有效性研究和最终的研究性新药（IND）使能研究，以支持人类 临床试验

PROJECT SUMMARY We are seeking funding to test the cardio protective effects of a novel new protein kinase C epsilon (PKCε) inhibitor (YT-002) and a novel protein kinase C beta II (PKCβII) inhibitor (YT-004) in the setting of porcine myocardial ischemia reperfusion (I/R) in vivo when given at the beginning of reperfusion. There currently is no approved pharmacological treatment/preventative for the I/R injury. We are fully aware of the past failures in this area. Literally all previous attempts have taken the oxygen free radical scavenging approach. We believe this underlies, at least in part, the reason there have been no successful pharmacological therapeutics for cardiac I/R injury to date. Once reactive oxygen species are generated the damage is immediate and scavenging is ineffective. Our approach – never been tried before – is to inhibit the generation of reactive oxygen species thus preventing their damage during reperfusion. Interviews we conducted with 22 interventional cardiologists overwhelmingly support the idea that such preservation of myocardial tissue and function would represent a significant long-term benefit to the patients experiencing ischemia-reperfusion injury leading to a decrease in the incidence of heart failure. Previous data from our lab with another compound YT-001 (identical mechanism of action as YT-002) demonstrated a significant inhibition of hydrogen peroxide generation in vivo, and a remarkable sparing of tissue damage and cardiac function post reperfusion in both the ex vivo rat I/R heart and the in vivo porcine myocardial I/R model. However, YT-001 has limited solubility and remaining patent life. Our new molecules (YT-002 and YT-004) have dramatically improved solubility, potency and new patent life. We have already demonstrated impressive infarct size reduction and cardiac function protection in isolated rat I/R hearts with YT-002 and YT-004. In this proposal, we will evaluate the cardio protective effects of YT-002 and YT-004 on ischemia reperfusion in the Gorman pig model (19). In addition to the animal’s vital signs, ejection fraction (measured by echocardiography) will be monitored (as an index of cardiac function/contractility) as well as cardiac proteins (e.g., troponin I) as an index of tissue damage throughout the protocol. Three hours after reperfusion, the hearts will be excised, and stained for direct assessment of cardiac damage (infarct size). Immunohistochemistry on frozen heart sections will be performed in Dr. Young’s lab at PCOM to detect PKCε and PKCβII localization. Based upon previously generated pig heart data with the PKCε inhibitor YT-001, we expect to see a dramatic in vivo preservation of cardiac tissue and function compared to animals receiving scrambled peptide control. These positive data will provide additional justification for performing longer-term animal efficacy studies and eventually Investigational New Drug (IND) enabling studies in support of human clinical trials.