Discovery of Potent 12-Lipoxygenase Inhibitors of Platelet Activation

发现有效的血小板激活 12-脂氧合酶抑制剂

基本信息

批准号：
8746993
负责人：
Theodore R Holman
金额：
$ 49.65万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8746993
关键词：
12-HETE Accounting Acute Adhesions Affect Age Anti-Inflammatory Agents Anti-inflammatory Arachidonate 12-Lipoxygenase Arachidonate 15-Lipoxygenase Arachidonate 5-Lipoxygenase Biochemical Biochemistry Biological Assay Biology Blood - brain barrier anatomy Blood Circulation Blood Platelets Blood Vessels Blood coagulation Cardiovascular Diseases Cells Characteristics Chemicals Chronic Clinical Coagulants Coagulation Process Collaborations Cytoplasmic Granules Data Development Disease Docking Drug Interactions Drug Kinetics Essential Fatty Acids Excretory function Failure Functional disorder Goals Grant Health Hemostatic function Heparin Human Hypertension In Vitro Individual Infection Inflammation Inflammation Process Inflammatory Injury Integrins Investigation Isoenzymes Laboratories Lipoxygenase Lipoxygenase 1 Lipoxygenase Inhibitors Liquid substance Metabolic Metabolic Clearance Rate Metabolism Modeling Monitor Morbidity - disease rate Mus Pathogenesis Pathway interactions Permeability Pharmaceutical Preparations Physiological Plasma Platelet Activation Platelet aggregation Play Prevention Process Production Property Prostaglandin-Endoperoxide Synthase Protein Binding Regulation Relative (related person)Research Risk Role Series Signaling Molecule Simulate Solubility Solutions Stroke Testing Therapeutic Thrombocytopenia Thrombosis Tissues Translations Tube United States National Institutes of Health Venous Thrombosis Wild Type Mouse Work absorption aqueous chemical property combat drug candidate drug development high throughput screening human disease in vivo in vivo Model inhibitor/antagonist lipid mediator mortality next generation novel novel strategies prevent receptor release of sequestered calcium ion into cytoplasm research study response risk benefit ratio therapeutic development tissue repair

项目摘要

DESCRIPTION (provided by applicant): The principle aim of this research is to develop selective/potent human 12-LOX inhibitors and probe the biochemistry of inflammation in platelet activation, leading to drug candidates for cardiovascular disease. Recent failures in testing of anti-coagulant agents have highlighted the need for new approaches to acute blood coagulation treatment. Among the novel targets, 12-lipoxygenase (12-LOX) stands out for numerous reasons: 1) It is highly expressed in platelets relative to other cells in circulation, 2) we have previously shown 12-LOX to be important for both hemostasis and thrombosis, and 3) 12-LOX is expressed in the platelets of both human and mice. The broad objective of the current proposal is to advance one or more candidate molecules toward therapeutic development against platelet activation/aggregation, utilizing our assays to identify novel selective inhibitor for the human 12-LOX. We have already performed a successful 12-LOX high-throughput (HTP) screen in collaboration with the NIH, which found over ~1000 potential hits. Of these ~1000 compounds, seven were found to be potent 12-LOX inhibitors. Two of these seven inhibitors were re- synthesized and shown to be potent and selective against human 12-LOX in vitro and effective in numerous human platelet cell assays, confirming them as "validated hits". Our research plan is guided by the following four specific aims. First, we propose to optimize our two "validated hits" and discover additional chemotypes, which have increased potency against human 12-LOX. Second, we shall optimize the ADME/PK properties of our "hits", maintain LOX isozyme selectivity and determine their mechanism of inhibition. Third, once we have optimized our various inhibitor chemotypes, we shall evaluate their protective potency in a variety of platelet activation screens. We will utilize these in vitro inhibitors in our platelet models and determine if individual inhibitors reduce aggregation, as well as ascertain if our inhibitors reduc the biochemical production of 12- HETE, as a direct marker of 12-LOX inhibition. Fourth, we will evaluate the in vivo efficacy of these 12-LOX inhibitors toward inhibition of thrombosis in mice while retaining normal levels of hemostasis. We will further confirm the activity of the 12-LOX inhibitors in vivo through assessment of 12-HETE production and ex vivo platelet function from mice treated with 12-LOX inhibitors. These inhibitors will additionally be tested for their abilityto prevent heparin-induced thrombocytopenia (HIT), as recent work suggests HIT is regulated in a 12- LOX-dependent manner. We are confident these studies will discover a 12-LOX inhibitor that is potentially an effective therapeutic against platelet aggregation in a number of pathological settings including athero- thrombosis, venous thrombosis, and HIT, and help us to better understand the biology of cardiovascular disease, a pathophysiological condition which worsens with age.

描述（由申请人提供）：这项研究的原理是开发选择性/有效的人类12-LOX抑制剂，并探测血小板激活中炎症的生物化学，从而导致候选候选心血管疾病的药物。抗凝蛋白剂测试的最新失败表明，需要对急性血液凝血治疗的新方法。在新的靶标中，12-脂氧合酶（12-lox）出于许多原因而脱颖而出：1）在血小板中相对于循环中的其他细胞，它在血小板中高度表达，2）我们先前已经显示12-lox对止血和血栓形成至关重要，而3）12-lox在人类和小鼠的血小板中表达。当前建议的广泛目的是将一个或多个候选分子推向针对血小板激活/聚集的治疗性发育，利用我们的测定方法来识别人类12-lox的新型选择性抑制剂。我们已经与NIH合作进行了成功的12-Lox高通量（HTP）屏幕，该屏幕发现超过1000次潜在的命中。在这些约1000种化合物中，发现7种是有效的12-lox抑制剂。这七个抑制剂中有两个被重新合成，并在体外对人类12-lox具有有效和选择性，并在许多人血小板细胞测定中有效，证实它们为“验证的命中”。我们的研究计划以以下四个特定目标为指导。首先，我们建议优化我们的两个“经过验证的命中”，并发现其他化学型，这增加了对人类12-Lox的效力。其次，我们将优化“命中”的ADME/PK特性，保持LOX同工酶的选择性并确定其抑制作用机制。第三，一旦我们优化了各种抑制剂化学型，我们将在各种血小板激活筛选中评估它们的保护效力。我们将在血小板模型中利用这些体外抑制剂，并确定单个抑制剂是否减少了聚集，并确定我们的抑制剂是否降低了12- HETE的生化产生，作为12-LOX抑制的直接标记。第四，我们将评估这些12-Lox抑制剂在小鼠中抑制血栓形成的体内功效，同时保留正常水平的止血水平。我们将通过评估用12-LOX抑制剂处理的小鼠的12-HETE产生和离体血小板功能，进一步确认体内12-LOX抑制剂的活性。这些抑制剂还将在预防肝素诱导的血小板减少症（HIT）的能力上进行测试，因为最近的工作表明，命中率是12个LOX依赖性方式进行调节。我们有信心这些研究将发现一种12-LOX抑制剂，该抑制剂在许多病理环境中可能是一种有效的针对血小板聚集的治疗方法，包括动脉粥样硬化，静脉血栓形成和命中，并帮助我们更好地了解心血管疾病的生物学，这是一种病理生理状况，随着年龄的增长。