BROAD-SPECTRUM RNAi THERAPEUTICS FOR FLAVIVIRAL ENCEPHALITIS

黄病毒性脑炎的广谱 RNAi 疗法

• 批准号: 7475259
• 负责人: MANJUNATH NARASIMHA SWAMY
• 金额: $ 84.9万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7475259
• 关键词: Acute; Address; Antiviral Agents; Arginine; Binding; Biological Assay; Biological Availability; Bioterrorism; Blood - brain barrier anatomy; Brain; Categories; Cells; Charge; Chemicals; Cholinergic Receptors; Clinical; Complex; Conserved Sequence; Culicidae; Data; Development; Drug Formulations; Effectiveness; Encephalitis; Ensure; Escape Mutant; Flavivirus; Frequencies; Gene Silencing; Genome; Genomics; Glycoproteins; Green Fluorescent Proteins; Half-Life; Hour; Human; In Vitro; Infection; Inflammatory; Injection of therapeutic agent; Interferons; Intravenous; Invasive; Japanese Encephalitis; Japanese encephalitis virus; Label; Lead; Liposomes; Localized; Mediating; Methods; Modification; Mus; National Institute of Allergy and Infectious Disease; Neurologic; Neurons; Numbers; Peptides; Peripheral; Pharmaceutical Preparations; Pharmacologic Substance; Property; RNA Binding; RNA Interference; Rabies virus; Range; Receptor Cell; Reporter Genes; Research Personnel; Route; Safety; Small Interfering RNA; Specificity; Subfamily lentivirinae; Technology; Testing; Therapeutic; Therapeutic Agents; Therapeutic Index; Therapeutic Uses; Time; Toxic effect; Toxicology; Viral; Viral Encephalitis; Virus; Virus Diseases; West Nile virus; base; brain cell; cell type; chemical property; design; disability; dosage; efficacy evaluation; immunogenicity; improved; in vivo; intravenous administration; intravenous injection; mortality; nervous system disorder; neurotropic; polyarginine; pre-clinical; programs; rabies virus glycoprotein G; receptor; response; small hairpin RNA; tool; transcytosis

DESCRIPTION (provided by applicant): West Nile (WN) and Japanese encephalitis (JE) viruses are flaviviruses that can cause a devastating acute neurological illness and currently there is no effective treatment for these viral infections. We have developed a RNA interference-based therapeutic method to suppress these viruses and shown the feasibility of using a single siRNA as a broad-spectrum antiviral agent to suppress fatal infections caused by both WNV and JEV in mice. However, the major challenge for harnessing the potential of siRNA for human therapy is the lack of suitable delivery methods. Delivery is particularly difficult in the CNS because of the presence of the blood-brain barrier. Preliminary studies suggest that a small peptide derived from the Rabies virus glycoprotein (RVG) may allow neuronal cell-specific targeting. Remarkably, when fused to a positively charged polymeric arginine peptide, the chimeric RVG-9R peptide was able to bind siRNA (by charge interaction) and deliver the siRNA to brain cells after intravenous injection in mice, resulting in specific gene silencing in the brain. Thus, we have identified a potential "siRNA drug" to treat flaviviral encephalitis as well as developed a clinically feasible method of delivering it to the brain by simple intravenous injection. In this proposal, in association with Alnylam Pharmaceuticals Inc, we will conduct preclinical evaluation of efficacy and safety, and advance the product development towards human therapy. Specifically, in aim1, we will conduct a comprehensive screen to select the most potent 2-3 lead antiviral siRNAs and chemically modify them for proper drug-like properties. In aim 2 we will address the bioavailability and safety/toxicology issues and determine the specificity of RNAi effects. In aim 3, we will test the efficacy of intravenous treatment to suppress encephalitis induced by JEV and WNV, both before and at different times after infection. To enhance the therapeutic index and safety of this treatment approach, in Aim 4 we will develop several different formulations of siRNA/peptide complex using RVG as the neuronal targeting agent. Relevance: These studies are expected to lead to the development of a viable RNAi-based treatment strategy for viral encephalitis. In addition, we would have developed a transvascular method for CNS delivery of siRNA and potentially other therapeutic agents that would facilitate treatment of a variety of neurological diseases.

描述（由申请人提供）： 西尼罗河 (WN) 和日本脑炎 (JE) 病毒属于黄病毒，可引起毁灭性的急性神经系统疾病，目前尚无针对这些病毒感染的有效治疗方法。我们开发了一种基于 RNA 干扰的治疗方法来抑制这些病毒，并证明了使用单一 siRNA 作为广谱抗病毒剂来抑制小鼠中由西尼罗河病毒和乙脑病毒引起的致命感染的可行性。然而，利用 siRNA 的潜力进行人类治疗的主要挑战是缺乏合适的递送方法。由于血脑屏障的存在，中枢神经系统中的递送尤其困难。初步研究表明，源自狂犬病病毒糖蛋白（RVG）的小肽可能允许神经元细胞特异性靶向。值得注意的是，当与带正电荷的聚合精氨酸肽融合时，嵌合RVG-9R肽能够结合siRNA（通过电荷相互作用），并在小鼠静脉注射后将siRNA递送至脑细胞，导致大脑中特定的基因沉默。因此，我们发现了一种潜在的“siRNA药物”来治疗黄病毒性脑炎，并开发了一种临床上可行的方法，通过简单的静脉注射将其输送到大脑。 在这项提案中，我们将与 Alnylam Pharmaceuticals Inc 合作，进行有效性和安全性的临床前评估，并推进针对人类治疗的产品开发。具体来说，在aim1中，我们将进行全面筛选，选择最有效的2-3先导抗病毒siRNA，并对它们进行化学修饰，以获得适当的药物样特性。在目标 2 中，我们将解决生物利用度和安全性/毒理学问题，并确定 RNAi 效应的特异性。在目标 3 中，我们将测试感染前和感染后不同时间静脉注射治疗抑制乙型脑炎病毒和西尼罗河病毒引起的脑炎的功效。为了提高这种治疗方法的治疗指数和安全性，在目标 4 中，我们将使用 RVG 作为神经元靶向剂开发几种不同的 siRNA/肽复合物配方。 关联： 这些研究预计将导致开发出一种可行的基于 RNAi 的病毒性脑炎治疗策略。此外，我们还开发了一种用于中枢神经系统递送 siRNA 的经血管方法，以及可能有助于治疗各种神经系统疾病的其他治疗药物。