Anti-tick-borne Encephalitis Virus Miniprotein Agents

抗蜱传脑炎病毒微蛋白制剂

• 批准号: 6932275
• 负责人: Robert O. Fox
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-15 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6932275
• 关键词: Flaviviridae; Tick borne encephalitis virus; West Nile virus; X ray crystallography; antiviral agents; bioterrorism /chemical warfare; cell surface receptors; combinatorial chemistry; conformation; electron microscopy; encephalitis virus; host organism interaction; laboratory mouse; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; phage display; protein binding; protein protein interaction; protein structure function; recombinant proteins; tissue /cell culture; virus envelope; virus protein

DESCRIPTION (provided by applicant): Our previous studies have provided a proof-of-principle that a small disulfide-rich miniprotein can be developed that will block the infection of cells by Langat (LGT) virus, a naturally attenuated virus that is a model for the pathogenic members of the tick-borne encephalitis (TBE) serogroup of the Flavivirus genus. A first generation miniprotein, termed MP-100, was selected by panning a conformationally restrained combinatorial miniprotein phage display library for binding with purified recombinant domain III (D3) of the LGT virus envelope (E) protein. The miniprotein MP-100 was shown to block infection of Vero and LLC-MK2 monkey kidney cell cultures by tick-borne LGT and Powassan viruses. Further studies indicated an antiviral effect in a mouse animal model. Our objective during this period of support is the development of a second-generation, more tightly binding miniprotein with improved antiviral activity against TBE serogroup flaviviruses compared to the current MP-100 sequence. Our goal is to develop an antiviral miniprotein that is effective against a broad range of potential flavivirus bioterrorist threat agents in the TBE serogroup, including Central European tick-borne encephalitis (strain Kumlinge), Kyasanur Forest Disease (FD), Omsk Hemorrhagic Fever (OHF) and Russian Spring Summer encephalitis (RSSE) viruses. We will identify optimized tight-binding analogs of MP-100 to OHF-E-D3 and the related TBE serogroup E-D3s, and determine if they have enhanced antiviral activity in tissue culture cells and in animals. The development of anti-TBE virus miniproteins will serve as a model for the development of miniproteins against other flaviviruses that are also potential bioterrorist threat agents or emerging diseases, including dengue, Japanese encephalitis and West Nile.

描述（由申请人提供）：我们之前的研究已经提供了原理证明，即可以开发出一种富含二硫键的小蛋白，该蛋白可以阻止 Langat (LGT) 病毒对细胞的感染，Langat (LGT) 病毒是一种自然减毒病毒，是一种模型黄病毒属蜱传脑炎 (TBE) 血清群的致病成员。第一代微型蛋白，称为 MP-100，是通过淘选构象限制的组合微型蛋白噬菌体展示文库来选择的，用于与 LGT 病毒包膜 (E) 蛋白的纯化重组结构域 III (D3) 结合。研究表明，小蛋白 MP-100 可以阻断蜱传 LGT 和 Powassan 病毒对 Vero 和 LLC-MK2 猴肾细胞培养物的感染。进一步的研究表明在小鼠动物模型中具有抗病毒作用。在此支持期间，我们的目标是开发第二代结合更紧密的微型蛋白，与当前的 MP-100 序列相比，其针对 TBE 血清群黄病毒的抗病毒活性有所提高。我们的目标是开发一种抗病毒小蛋白，可有效对抗 TBE 血清群中多种潜在的黄病毒生物恐怖威胁因子，包括中欧蜱传脑炎（Kumlinge 株）、Kyasanur 森林病 (FD)、鄂木斯克出血热 (OHF) ）和俄罗斯春夏脑炎（RSSE）病毒。我们将鉴定 MP-100 与 OHF-E-D3 和相关 TBE 血清群 E-D3 的优化紧密结合类似物，并确定它们在组织培养细胞和动物中是否具有增强的抗病毒活性。抗TBE病毒小蛋白的开发将作为开发针对其他黄病毒的小蛋白的模型，这些黄病毒也是潜在的生物恐怖威胁因子或新出现的疾病，包括登革热、日本脑炎和西尼罗河病毒。