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利用三维 结构数据来设计抗病毒药物。 我们的长远目标是 三个方面：1）建立药物靶点结构数据库 复合物，2）开发使用这些结构的药物 设计，以及3）设计具有改善的活性或拓宽的活性的新型药物。 活动范围。 这些药物将是相当大的医疗和 预防鼻病毒性感冒的经济意义 （在美国，感冒占80%）， 对小儿麻痹症和柯萨奇病毒引起的疾病进行早期干预。 此外，由于广泛的拟议目标的相似性， 一系列RNA病毒，这些研究中确定的药物可以作为 用于开发抗病毒药物的有用先导化合物， 各种其他重大人类和兽医疾病。 初步研究将集中在晶体学研究之间的复合物 脊髓灰质炎病毒和一系列充分表征的化合物（其中几种 正在作为抗鼻病毒剂进入试验）， 生产性细胞附着或细胞进入。 晶体学研究 将包括杨森制药公司生产的抗病毒药物的复合物 Sterling-Winthrop用3型Sabin株和Mahoney株 1型脊髓灰质炎病毒株。 晶体学研究也将 包括耐药变体的结构（将选择 并由威斯康星州大学的罗兰吕克特测序）。 的 配合物的结构将与 与鼻病毒14和鼻病毒1a的类似试剂的复合物（目前 Rossmann et.在普渡大学）。 的信息 从结构中得到的信息将用于设计和合成化合物 活动和特异性改变（与杨森合作 制药公司和K.C. RISC的Nicolau）。 随着这些研究的进展，我们将扩大该计划，包括额外的 药物设计的目标，从脊髓灰质炎病毒RNA依赖的RNA开始， 聚合酶（3Dpol）和负责切割衣壳的蛋白酶 蛋白质前体（3CD）。 大量高纯度 蛋白质已被鉴定为3D和3CD。 3D的样本在 早期阶段的结晶屏幕，和样品的3CD是预期的 在拟议供资期开始之前。 我们建议 产生这些酶的晶体，以解决游离的 酶和酶抑制剂复合物，并利用这种结构 信息，以及从设计中获得的经验， 去包被抑制剂，以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.