Anti-tick-borne Encephalitis Virus Miniprotein Agents

描述（由申请方提供）：我们之前的研究提供了一个原理证明，即可以开发一种富含二硫化物的小微蛋白，其将阻断Langat（LGT）病毒对细胞的感染，Langat（LGT）病毒是一种天然减毒病毒，是黄病毒属蜱传脑炎（TBE）血清群致病成员的模型。通过淘选构象受限的组合微蛋白噬菌体展示文库以与LGT病毒包膜（E）蛋白的纯化重组结构域III（D3）结合来选择第一代微蛋白，称为MP-100。微蛋白MP-100可阻断蜱传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，与目前的MP-100序列相比，对TBE血清群黄病毒具有更高抗病毒活性的更紧密结合的微蛋白。我们的目标是开发一种抗病毒微蛋白，可有效对抗TBE血清群中广泛的潜在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病毒微蛋白的开发将作为开发针对其他黄病毒的微蛋白的模型，这些黄病毒也是潜在的生物恐怖主义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。

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.