Prevention of Cardiac Cell Death by Mst1 inhibitor

描述(申请人提供)：6.摘要美国每年有近2000万人患有缺血性心脏病，100万患者发生心肌梗死(MI)。许多患者即使在急性事件中幸存下来也会出现心力衰竭，这清楚地表明目前的干预措施是不够的。因此，开发一类新的预防缺血/再灌注(I/R)损伤的药物将具有巨大的市场机会和重大的临床进展。心肌细胞死亡，包括细胞凋亡，与心力衰竭的发病密切相关。哺乳动物不育20样激酶1(Mst1)在介导心肌细胞凋亡中发挥重要作用，并与充血性心力衰竭的发病密切相关(Vasade，美国专利7160859,2007年)。利用心脏特异表达显性负性Mst1的转基因小鼠(Tg-dN-Mst1)，抑制内源性Mst1可抑制I/R后心肌细胞的凋亡，从而缩小MI的大小，改善长期心功能。Vasade的长期目标之一是开发Mst1特异性抑制剂用于治疗I/R损伤。通过我们获得美国国立卫生研究院第一阶段STTR拨款的努力，Vasade 1)发现了一种新型的特异性多肽Mst1抑制剂(称为Mst1抑制肽120(MIP120))，并使其成为细胞通透性(称为TAT120)；2)建立了一种基于ELISA的高通量方法，可以准确测定Mst1的激酶活性。据我们所知，MIP120和TAT120是目前仅有的抑制Mst1激酶活性的化合物。在第二阶段STTR项目中，Vasade将提供明确的证据，表明Mst1抑制剂在体内有助于防止I/R引起的心肌细胞死亡，并确定一种精制的先导化合物，用于开发用于治疗I/R损伤的多肽模拟Mst1抑制剂。目的：1确定TAT120对在体心肌缺血再灌注损伤大小和心肌细胞凋亡的影响，为应用Mst1抑制剂减轻缺血再灌注损伤奠定理论基础。目的2为进一步完善MIP120/TAT120作为Mst1抑制剂的性质，阐明MIP120/TAT120的结构-功能关系，为开发更好的多肽或多肽模拟物提供有力的概念证明和精制先导化合物，为进一步开发多肽模拟小分子Mst1抑制剂用于心脏I/R损伤的临床治疗提供有力的证据。 公共卫生相关性：美国每年有近2000万人患有缺血性心脏病，100万患者发展为心肌梗死(MI)。许多患者即使在急性事件中幸存下来也会发生心力衰竭，这清楚地表明，目前的干预措施不足以防止心肌细胞死亡和由此导致的左心室(LV)功能障碍。目前的研究目标是开发一种新的预防缺血/再灌注(I/R)损伤的药物，该药物应该具有很大的市场机会和重要的临床意义。

DESCRIPTION (provided by applicant): 6. Abstract Almost 20 million people suffer from ischemic heart diseases in US and one million patients develop myocardial infarction (MI) every year. Many patients develop heart failure even if they survive the acute event, which clearly indicates that current interventions are not sufficient. Thus, the development of a new class of medicine, which prevents ischemia/reperfusion (I /R) injury, would have a large market opportunity and significant clinical advance. Cardiac myocyte death, including apoptosis, is intimately involved in the pathogenesis of heart failure. Mammalian sterile 20 like kinase 1 (Mst1) plays an important role in mediating cardiac myocyte apoptosis and is intimately involved in the pathogenesis of congestive heart failure (Vasade, US patent 7160859, 2007). Using transgenic mice with cardiac specific overexpression of dominant negative Mst1 (Tg-DN-Mst1), inhibition of endogenous Mst1 suppresses cardiac myocyte apoptosis in response to I/R, thereby reducing the size of MI and improving long-term cardiac function. One of the long-term goals of Vasade is to develop Mst1 specific inhibitors for the treatment of I/R injury. Through our effort made possible by the phase I STTR grant from NIH, Vasade 1) has discovered a novel and specific peptide Mst1 inhibitor (termed Mst1 inhibitory peptide 120 (MIP120)) and made it cell permeable (termed TAT120) and 2) has established an ELISA-based high throughput method to accurately determine the kinase activity of Mst1. To our knowledge, MIP120 and TAT120 are the only compounds currently available inhibiting the kinase activity of Mst1. In the phase II STTR project, Vasade will provide unequivocal evidence showing that the Mst1 inhibitor is useful for the prevention of cardiac myocyte death by I/R in vivo and identify a refined lead compound for the development of peptide mimetic Mst1 inhibitors for treatment of I/R injury. Specific aims are: Aim 1 To determine the effect of TAT120 on the size of MI and cardiac myocyte apoptosis in response to I/R in vivo, thereby establishing the proof of concept for the use of Mst1 inhibitors to reduce I/R injury using the mouse and pig models of ischemia/reperfusion. Aim 2 To further improve the property of MIP120 /TAT120 as Mst1 inhibitors and to elucidate the structure- function relationship of MIP120/TAT120 for the development of better peptides or peptide mimetics This phase II study will provide strong proof of concept and refined lead compounds, which will be used for the development of peptide mimetic small molecule Mst1 inhibitors for the future clinical treatment of I/R injury in the heart. PUBLIC HEALTH RELEVANCE: Almost 20 million people suffer from ischemic heart diseases in US and one million patients develop myocardial infarction (MI) every year. Many patients develop heart failure even if they survive the acute event, which clearly indicates that current interventions are not sufficient to prevent myocardial cell death and resultant left ventricular (LV) dysfunction. The current investigation is aiming at generating a new class of medicine preventing ischemia/reperfusion (I /R) injury, which should have a large market opportunity and significant clinical importance.