Anti-tick-borne Encephalitis Virus Miniprotein Agents

描述(申请人提供)：我们之前的研究已经提供了一个原则证明，即可以开发出一种富含二硫键的小蛋白，可以阻止兰加特(LGT)病毒对细胞的感染，兰加特(LGT)病毒是一种自然减毒病毒，是黄病毒属森林脑炎(TBE)血清组致病成员的模型。通过筛选构象受限的组合小蛋白噬菌体展示文库，筛选出一种命名为MP-100的第一代小蛋白，用于与纯化的LGT病毒囊膜蛋白的重组结构域III(D3)结合。微蛋白MP-100能有效地阻断森林病毒和鲍瓦桑病毒感染培养的Vero和LLC-MK2猴肾细胞。进一步的研究表明，在小鼠动物模型中有抗病毒作用。在这一支持期间，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，具有更好的抗TBE血清群黄病毒活性。我们的目标是开发一种抗病毒微型蛋白，可有效对抗TBE血清组中一系列潜在的黄病毒生物恐怖威胁因素，包括中欧森林脑炎(Kumlinger株)、凯萨努尔森林病(FD)、鄂木斯克出血热(OHF)和俄罗斯春夏脑炎(RSSE)病毒。我们将确定优化的MP-100到OHF-E-D3的紧密结合类似物和相关的TBE血清组E-D3S，并确定它们是否在组织培养细胞和动物中具有增强的抗病毒活性。抗TBE病毒微型蛋白的开发将作为开发针对其他黄病毒的微型蛋白的模型，其他黄病毒也是潜在的生物恐怖威胁因素或新出现的疾病，包括登革热、日本脑炎和西尼罗河。

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.