A new Chinese herb-derived selective Toll-like receptor antagonist (Project 1)

一种新型中药选择性Toll样受体拮抗剂（项目1）

基本信息

批准号：
8460786
负责人：
Daping Fan
金额：
$ 20.55万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
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 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8460786
关键词：
Adaptor Signaling Protein Adhesions Anti-Inflammatory Agents Anti-inflammatory Antiatherogenic Arterial Fatty Streak Atherosclerosis Attenuated Binding Blood Vessels Cells China Chinese Herbs Chinese Traditional Medicine Chronic Clinic Clinical Trials Complementary and alternative medicine Data Deposition Development Diet Disease Endothelial Cells Fatty acid glycerol esters Herb Human In Vitro Infiltration Inflammation Inflammatory Inflammatory Response Interleukin-1 Invaded Investigation Leukocytes Ligands Lipids Lipopolysaccharides Low Density Lipoprotein Receptor Measures Mediating Molecular Mus Myocardial Infarction Natural Immunity Pharmacology Phenotype Plants Play Proteins Role Signal Transduction Smooth Muscle Myocytes Staging Stroke TLR2 gene TLR3 gene TLR4 gene Testing Therapeutic Therapeutic Agents Tissues Toll-like receptors United States Vascular Endothelial Cell Viral Work abstracting atherogenesis base cell motility dietary supplements driving force feeding hypercholesterolemia immune function inflammatory marker macrophage monocyte novel oxidized lipid pathogen receptor vascular inflammation

项目摘要

Abstract: Atherosclerosis is the direct cause of heart attack and stroke, the No. 1 and No. 3 killers in the United States, respectively. It is the result of both lipid deposition and chronic vascular inflammation. Toll-like receptors (TLRs), the key components of innate immunity, play detrimental roles in every stage of atherosclerosis, with TLR2 and TLR4 being best documented. While participating in the first line of defense against invading pathogens, TLR2- and TLR4-mediated signaling is considered to be a driving force in atherogenesis. Thus, blockade of TLR2 and TLR4 signaling is an intriguing therapeutic approach for atherosclerosis. However, no TLR2 or TLR4 antagonists are currently approved for clinic use. Recently, in an effort to isolate and characterize single compounds from Sparganium stoloniferum tubers, a commonly used Traditional Chinese Medicine herb, we obtained a novel compound, designated Sparstolonin B (SsnB), and made exciting discoveries. Our preliminary studies show that 1) SsnB has potent anti-inflammatory effects on macrophages by selectively blocking TLR2 and TLR4 signaling; 2) SsnB diminishes the ability of activated endothelial cells to attract monocytes for adhesion, and decreases arterial smooth muscle cell migration; and 3) SsnB effectively suppresses inflammatory response to lipopolysaccharide (LPS) in mice. On the basis of these preliminary data, we hypothesize that SsnB can be developed as an anti-inflammatory and anti-atherogenic agent by virtue of its selective inhibitory effects on TLR2 and TLR4 signaling. To test this hypothesis, we propose three specific aims. SA1. To elucidate the molecular mechanism by which SsnB blocks TLR2 and TLR4 signaling. We will express and purify the Toll/IL-1 receptor (TIR) domains of TLRs, the adaptor proteins TIRAP/Mal and MyD88, and examine the binding of SsnB to these proteins. SA2. To examine the effects of SsnB on resident vascular cells. We will test the hypothesis that SsnB suppresses the inflammatory phenotype in arterial endothelial and smooth muscle cells by blocking TLR2 and TLR4 signaling. SA3. To test the hypothesis that SsnB attenuates atherogenesis in mice. LDL receptor (LDLR) deficient mice will be fed high fat diet to induce hypercholesterolemia and atherosclerosis. SsnB will be administrated to test if it attenuates atherogenesis in these mice. In summary, this study will test the anti-inflammatory and anti-atherogenic effects of a new natural compound recently isolated and characterized by us. The confirmation of the hypothesis that SsnB has anti-inflammatory and anti-atherogenic merit by blocking TLR2 and TLR4 signaling in macrophages and resident vascular cells will usher the development of this compound into an anti-atherogenic agent. This study will also provide a new example of complementary and alternative medicine bridging with modern pharmacology.

抽象的：动脉粥样硬化是心脏病和中风的直接原因，分别是美国第一和第三号杀手。它是脂质沉积和慢性血管炎症的结果。 Toll 样受体 (TLR) 是先天免疫的关键组成部分，在动脉粥样硬化的每个阶段都发挥着有害作用，其中 TLR2 和 TLR4 得到了最好的记录。在参与抵御入侵病原体的第一道防线时，TLR2 和 TLR4 介导的信号传导被认为是动脉粥样硬化形成的驱动力。因此，阻断 TLR2 和 TLR4 信号传导是动脉粥样硬化的一种有趣的治疗方法。然而，目前还没有 TLR2 或 TLR4 拮抗剂被批准用于临床。最近，为了从常用中药草三棱块茎中分离和表征单一化合物，我们获得了一种新化合物，命名为三棱草素 B (SsnB)，并取得了令人兴奋的发现。我们的初步研究表明：1）SsnB 通过选择性阻断 TLR2 和 TLR4 信号传导，对巨噬细胞具有有效的抗炎作用； 2) SsnB 降低活化的内皮细胞吸引单核细胞粘附的能力，并减少动脉平滑肌细胞的迁移； 3) SsnB 有效抑制小鼠对脂多糖 (LPS) 的炎症反应。基于这些初步数据，我们假设 SsnB 凭借其对 TLR2 和 TLR4 信号传导的选择性抑制作用，可以开发为抗炎和抗动脉粥样硬化药物。为了检验这一假设，我们提出了三个具体目标。 SA1。阐明 SsnB 阻断 TLR2 和 TLR4 信号传导的分子机制。我们将表达和纯化 TLR 的 Toll/IL-1 受体 (TIR) 结构域、接头蛋白 TIRAP/Mal 和 MyD88，并检查 SsnB 与这些蛋白的结合。 SA2。检查 SsnB 对常驻血管细胞的影响。我们将检验 SsnB 通过阻断 TLR2 和 TLR4 信号传导来抑制动脉内皮和平滑肌细胞炎症表型的假设。 SA3。测试 SsnB 减弱小鼠动脉粥样硬化形成的假设。低密度脂蛋白受体（LDLR）缺陷的小鼠将被喂食高脂肪饮食以诱发高胆固醇血症和动脉粥样硬化。将施用 SsnB 来测试它是否能减轻这些小鼠的动脉粥样硬化形成。总之，本研究将测试我们最近分离和表征的一种新天然化合物的抗炎和抗动脉粥样硬化作用。 SsnB 通过阻断巨噬细胞和常驻血管细胞中的 TLR2 和 TLR4 信号传导而具有抗炎和抗动脉粥样硬化功效的假设得到证实，这将推动该化合物发展成为抗动脉粥样硬化剂。这项研究还将提供补充和替代医学与现代药理学相结合的新例子。