Multicenter preclinical trial of rho-kinase inhibitor fasudil in acute focal cerebral ischemia and reperfusion

Rho激酶抑制剂法舒地尔治疗急性局灶性脑缺血再灌注的多中心临床前试验

• 批准号: 10246264
• 负责人: Cenk Ayata
• 金额: $ 53.08万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246264
• 关键词: Actins; Acute; Adhesions; Advocate; Alteplase; Anti-Inflammatory Agents; Apoptosis; Aspirin; Blood - brain barrier anatomy; Blood Cells; Blood Vessels; Blood Viscosity; Brain Edema; Cardiovascular system; Carotid Atherosclerotic Disease; Cerebral Infarction; Cerebral Ischemia; Cerebrovascular Circulation; China; Chronic; Clinical; Clinical Trials; Coagulation Process; Common Data Element; Complement; Core-Binding Factor; Coronary Arteriosclerosis; Cyclic AMP-Dependent Protein Kinases; Cytoskeleton; Diabetes Mellitus; Dose; Edema; Endothelium; Experimental Models; Failure; Female; Filament; Formulation; Functional disorder; Heart failure; Hemorrhage; Hypertension; Imaging Device; Inbred SHR Rats; Infarction; Inflammation; Intervention; Ischemia; Ischemic Stroke; Japan; Leukocytes; Magnetic Resonance Imaging; Manuscripts; Middle Cerebral Artery Occlusion; Modeling; Mus; Neurocognitive; Neuroglia; Neurologic; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Outcome Assessment; Patients; Pharmaceutical Preparations; Pharmacology; Physiological; Platelet aggregation; Preclinical Testing; Process; Protein Kinase; Publishing; Pulmonary Hypertension; Rattus; Recording of previous events; Reperfusion Therapy; Research Personnel; Rho-associated kinase; Safety; Sample Size; Scientist; Sensorimotor functions; Series; Serious Adverse Event; Stroke; Subarachnoid Hemorrhage; Subgroup; Swelling; Testing; Therapeutic; Tissues; Vascular Smooth Muscle; Vasodilation; acute care; acute stroke; aged; atorvastatin; blood-brain barrier disruption; cerebrovascular; clinically relevant; comorbidity; db/db mouse; design; efficacy evaluation; efficacy testing; efficacy trial; environmental enrichment for laboratory animals; excitotoxicity; experience; experimental study; fasudil; functional outcomes; healthy volunteer; hypercholesterolemia; improved; improved outcome; indexing; inhibitor/antagonist; ischemic injury; kinase inhibitor; male; molecular marker; mortality; natural hypothermia; novel; novel therapeutics; off-patent; optical imaging; pre-clinical assessment; preclinical trial; protein kinase inhibitor; research clinical testing; response; rho; safety testing; sex; standard of care; stroke model; stroke patient; stroke therapy; support network; systematic review; thrombolysis; tool; treatment effect

In this preclinical trial, we propose to target rho-associated protein kinase (ROCK) using fasudil in models of acute focal cerebral ischemia followed by reperfusion. ROCK is a major regulator of actin cytoskeleton controlling numerous functions in vascular smooth muscle, endothelium, neurons, glia, leukocytes and blood cells. Many of these are relevant for the pathophysiology of stroke, making ROCK a unique pleiotropic target with multiple converging and synergistic mechanisms, including cerebrovasodilation and reduced blood viscosity improving collateral flow in hyperacute stroke, and anti-inflammatory and anti-edema effects in acute to subacute stages. We and others have targeted ROCK in models of ischemic stroke in exploratory studies, vast majority of which showed improved tissue and functional outcomes, in multiple species and stroke models, and using various inhibitors and therapeutic paradigms. In a systematic review and stratified metaanalysis of 25 experimental studies, ROCK inhibition reduced infarct volume by 37% and improved neurological scores by 41%. Fasudil is the most commonly used ROCK inhibitor in experimental stroke and is off-patent to be procured in any formulation for both preclinical and clinical trials without delay. Fasudil has been the clinical standard of care for two decades in acute SAH in China and Japan, with a clean safety record. There have been numerous clinical trials of systemic fasudil in both US and abroad in healthy volunteers, and in patients with chronic cerebral infarcts, coronary artery disease, heart failure, and pulmonary hypertension among others. Indeed, in a small trial in acute stroke fasudil was efficacious. Nevertheless, none of these trials revealed any serious adverse events either with acute single doses or chronic daily dosing. We propose to confirm this promising profile in a preclinical trial using transient middle cerebral artery occlusion. Aim 1 will establish the optimal dose (Exp 1), confirm efficacy on long-term outcome (Exp 2), test whether fasudil extends the therapeutic window for recanalization (Exp 3), examine the efficacy of intraarterial delivery (Exp 4), and confirm efficacy using clinically-relevant MRI indices (Exp 5). Aim 2 will then test efficacy and safety in a clot model with or without tPA (Exp 6), and safety in comorbid hypertension and diabetes as well as combination treatments (Exp 7) and in permanent ischemia (Exp 8). The proposal comes from an experienced group of investigators with proven expertise, capable of adjusting to the needs of the network. Collectively, we have published ~100 manuscripts using various stroke models and neurocognitive and tissue readouts suitable for both acute (<72h) and chronic (up to 2 months) assessments in mice and rats. Besides the proposed experimental models, our expertise covers hemorrhagic transformation, combination treatment with tPA and ischemic brain edema. Altogether, we passionately support the concept of an unbiased multicenter preclinical testing platform such as SPAN for experimental stroke therapeutics. We are fully committed to support this network.

在这项临床前试验中，我们建议在急性局灶性脑缺血再灌注模型中使用法舒地尔靶向 rho 相关蛋白激酶 (ROCK)。 ROCK 是肌动蛋白细胞骨架的主要调节因子，控制血管平滑肌、内皮、神经元、神经胶质、白细胞和血细胞的多种功能。其中许多与中风的病理生理学相关，使 ROCK 成为独特的多效性靶点，具有多种汇聚和协同机制，包括脑血管舒张和降低血液粘度，改善超急性中风的侧支血流，以及急性至亚急性阶段的抗炎和抗水肿作用。我们和其他人在探索性研究中针对缺血性中风模型中的 ROCK 进行了靶向研究，其中绝大多数研究在多个物种和中风模型中显示出组织和功能结果的改善，并使用了各种抑制剂和治疗范例。在对 25 项实验研究的系统回顾和分层荟萃分析中，ROCK 抑制使梗塞体积减少了 37%，神经系统评分改善了 41%。 Fasudil 是实验性中风中最常用的 ROCK 抑制剂，并且已过专利，可以立即以任何制剂形式采购用于临床前和临床试验。二十年来，法舒地尔一直是中国和日本急性蛛网膜下腔出血的临床标准治疗，具有良好的安全记录。美国和国外已在健康志愿者以及慢性脑梗塞、冠状动脉疾病、心力衰竭和肺动脉高压等患者中进行了大量全身法舒地尔临床试验。事实上，在一项针对急性中风的小型试验中，法舒地尔是有效的。然而，这些试验均未发现急性单剂量或长期每日给药有任何严重不良事件。我们建议在一项使用短暂大脑中动脉闭塞的临床前试验中证实这一有希望的情况。目标 1 将确定最佳剂量（实验 1），确认长期结果的疗效（实验 2），测试法舒地尔是否延长再通治疗窗（实验 3），检查动脉内给药的疗效（实验 4），并使用临床相关 MRI 指数确认疗效（实验 5）。然后，目标 2 将测试使用或不使用 tPA 的血栓模型（实验 6）的有效性和安全性，以及合并高血压和糖尿病以及联合治疗（实验 7）和永久性缺血（实验 8）的安全性。该提案来自经验丰富的研究人员小组，他们拥有经过验证的专业知识，能够根据网络的需求进行调整。我们总共发表了约 100 篇手稿，使用各种中风模型以及神经认知和组织读数，适用于小鼠和大鼠的急性（<72 小时）和慢性（长达 2 个月）评估。除了提出的实验模型外，我们的专业知识还包括出血转化、tPA 联合治疗和缺血性脑水肿。总而言之，我们热情支持公正的多中心临床前测试平台的概念，例如用于实验性中风治疗的 SPAN。我们完全致力于支持这个网络。