POLIOVIRUS AS A MODEL FOR THE DESIGN OF ANTIVIRAL DRUGS

我们建议使用脊髓灰质炎病毒作为一个范例，利用三维 用于设计抗病毒药物的结构数据。我们的长期目标将是 三：1)建立药物靶点结构数据库 络合物，2)开发将这些结构用于药物的方案 设计，以及3)设计活性更高或范围更广的新药 活动的光谱。这些药物将具有相当大的医疗和医疗价值 预防鼻病毒引起的感冒的经济意义 (它们占美国感冒的80%)和 对脊髓灰质炎和柯萨奇病毒所致疾病的早期干预。 此外，由于拟议的目标在广泛的范围内都有相似之处 一系列RNA病毒，这些研究中确定的药物可以用作 用于开发治疗疟疾的抗病毒药物的有用先导化合物 各种各样的其他重大人类和兽医疾病。 初步研究将集中在配合物的结晶学研究上。 脊髓灰质炎病毒和一系列具有良好特性的化合物(其中几个 正在作为抗鼻病毒药物进入试验)，它们结合病毒粒子并阻止 生产性细胞附着或细胞进入。结晶学研究 将包括Janssen PharmPharmtica生产的抗病毒药物复合体 和斯特林-温斯罗普与萨宾类型3和马奥尼品系 1型脊髓灰质炎病毒株。结晶学研究还将 包括耐药变异体的结构(将被选择 由威斯康星大学的罗兰·鲁克特测序)。这个 配合物的结构将与化合物的结构进行比较。 类似药物与鼻病毒14和鼻病毒1a的复合体(目前 正在由Rossmann et研究。阿尔在普渡大学)。这些信息 从这些结构中获得的物质将被用于设计和合成化合物 更改活动和细节(与Janssen合作 PharmPharmtica和K.C.Nicolau在RISC)。 随着这些研究的进展，我们将扩大该计划，以包括其他 药物设计的目标，从依赖脊髓灰质炎病毒RNA的RNA开始- 聚合酶(3Dpoll)与衣壳裂解相关的蛋白酶 蛋白质前体(3CD)。大量高纯度物质的来源 蛋白质在3D和3CD中都已被鉴定。3D的样本在 结晶屏的早期阶段，预计将有3CD样品 在拟议的资助期开始之前。我们建议 产生这些酶的晶体，以解决游离态的结构 酶和酶-抑制物复合体，并利用这种结构 信息，以及从设计中获得的经验 揭开抑制剂的外衣，设计具有抗病毒活性的化合物。

We propose to use poliovirus as a paradigm for utilizing three-dimensional structural data to design antiviral drugs. Our long range goals will be threefold: 1) to develop a data base of structures of drug-target complexes, 2) to develop protocols for using these structures for drug design, and 3) to design novel drugs with improved activity or a broadened spectrum of activity. These drugs would be of considerable medical and economic significance for prophylaxis of rhinovirus induced common cold (which account for up to 80% of the colds in the United States) and for early intervention in poliomyelitis and coxsackievirus induced diseases. In addition, because of similarities in the proposed targets across a wide range of RNA viruses, the drugs identified in these studies could serve as useful lead compounds for the development of antivirals for treatment of a wide variety of other significant human and veterinary diseases. Initial studies will focus on crystallographic studies of complexes between poliovirus and a series of well-characterized compounds (several of which are entering trials as antirhinovirus agents) which bind virions and block productive cell attachment or cell entry. The crystallographic studies will include complexes of antiviral agents made by Janssen Pharmaceutica and by Sterling-Winthrop with the Sabin strain of type 3 and the Mahoney strain of type 1 poliovirus. The crystallographic studies will also include the structures of drug resistant variants (which will be selected and sequenced by Roland Rueckert at the University of Wisconsin). The structures of the complexes will be compared with the structures of complexes of similar agents with rhinovirus 14 and rhinovirus 1a (currently being studied by Rossmann et. al at Purdue University). The information gained from the structures will be used to design and synthesize compounds with altered activities and specificities (in collaboration with Janssen Pharmaceutica and with K.C. Nicolau at RISC). As these studies progress we will expand the program to include additional targets for drug design, beginning with the poliovirus RNA-dependent RNA- polymerase (3Dpol) and the protease responsible for the cleavage of capsid protein precursors (3CD). Sources of large amounts of highly purified protein have been identified for both 3D and 3CD. Samples of 3D are in the early phase of crystallization screens, and samples of 3CD are expected prior to the beginning of the proposed funding period. We propose to produce crystals of these enzymes, to solve the structure of the free enzymes, and enzyme-inhibitor complexes, and to utilize this structural information, together with the experience gained from the design of uncoating inhibitors, to design compounds with antiviral activity.