Antiviral Drugs for Treatment of Herpes B Infections

描述（由申请人提供）：猴B型疱疹病毒（BV）已被报道在人类中引起致命感染。BV原产于研究用猕猴，是一种潜在的B类生物恐怖制剂。人类疾病的特征是皮肤或粘膜上出现水疱状疹，与1型单纯疱疹病毒或2型单纯疱疹病毒引起的疱疹性病变难以区分。不像典型的HSV感染很少导致中枢神经系统受损伤，BV引起迅速上升的脊髓炎和脑炎，几乎总是导致死亡。由于对处理人员的威胁和用于生物恐怖主义的潜力，BV感染的诊断和治疗需要改进。我们建议开发新的乙肝抗病毒靶点，包括克隆选定的DNA复制相关基因，表达和分离蛋白质产物，以及酶的底物和抑制剂特性的表征。我们的专业知识和广泛的抑制HSV酶的化合物文库将成为药物发现的起点。由于人类BV感染的病理性质，我们将在这个I期项目的初始努力集中在胸苷激酶（TK）上，因为HSV 1型和2型TK的抑制剂可以保护小鼠免受致死性脑炎的侵害。具体目的是：1)克隆并在真核表达载体上过表达BV TK基因，纯化足够量的酶供研究；2)全面表征bvtk的酶学特性；3)筛选对HSV TKs具有广泛亲和力的n2 -苯鸟嘌呤衍生物文库；4)合成新的类似物以提高效价和选择性；5)在体外细胞培养中测试有前途的化合物和抗疱疹药物的抗细菌性疱疹活性。

DESCRIPTION (provided by applicant): Monkey Herpes B virus (BV) has been reported to cause lethal infections in humans. BV is indigenous to macaque monkeys used in research, and is a potential group B bioterrorism agent. Human disease is characterized by vesicular eruptions on the skin or mucous membranes that are indistinguishable from herpetic lesions caused by HSV-1 or HSV-2. Unlike typical HSV infections that rarely lead to central nervous system involvement, BV causes a rapidly ascending myelitis and encephalitis that almost always leads to death. Because of the threat to handlers and potential for use in bioterrorism, diagnosis and treatment of BV infection need to be improved. We propose to develop new antiviral targets for HB involving cloning of selected DNA replication-related genes, expression and isolation of the protein products, and characterization of the substrate and inhibitor properties of the enzymes. Our expertise and extensive libraries of compounds that inhibit the HSV enzymes will be the starting point in drug discovery. Because of the nature of the pathology of BV infection in humans, we will focus initial efforts in this phase I program on thymidine kinase (TK), because inhibitors of the HSV types 1 and 2 TKs protect mice from lethal encephalitis. The specific aims are: 1) to clone and overexpress the BV TK gene in eukaryotic expression vectors, and purify sufficient quantities of the enzyme for study; 2) to fully characterize the enzymatic properties of BV TK; 3) to screen a library of N2-phenylguanine derivatives that have a wide range of affinities for the HSV TKs; 4) to synthesize new analogs to improve potency and selectivity; and 5) to test promising compounds and antiherpes drugs in vitro for anti-BV activity in cell cultures.