Boehringer Ingelheim Ltd, Canada.
Curr Top Microbiol Immunol. 2011;348:61-88. doi: 10.1007/82_2010_92.
Human papillomaviruses are responsible for multiple human diseases, including cervical cancer caused by multiple high-risk types and genital warts caused by the low-risk types 6 and 11. Based on the research indicating that low-risk HPV could be successfully targeted by inhibitors of viral DNA replication, we carried out several high-throughput screens for inhibitors of DNA replication activities. Two series were identified in screens for inhibitors of the interaction between the viral proteins E1 and E2. The two series were demonstrated to bind to overlapping sites on the transactivation domain of E2, at the E1-binding interface, by a series of biochemical and biophysical experiments. A member of the first series was also cocrystallized with the E2 transactivation domain. For both series, structure-activity investigations are described, which resulted in several hundred fold improvements in activity. The best compounds in each series had low nanomolar activity against the HPV11 E1-E2 interaction, and EC(50) values in cellular DNA replication assays of approximately 1 μM. Binding modes for the two series are compared, and some general conclusions about the discovery of protein-protein interaction inhibitors are drawn from the work described.
人乳头瘤病毒可引起多种人类疾病，包括由多种高危型引起的宫颈癌和由低危型 6 和 11 引起的生殖器疣。基于研究表明，低危型 HPV 可以被病毒 DNA 复制抑制剂靶向，我们进行了多次针对 DNA 复制活性抑制剂的高通量筛选。在针对病毒蛋白 E1 和 E2 相互作用抑制剂的筛选中发现了两个系列。通过一系列生化和生物物理实验证明，这两个系列都结合在 E2 的转录激活域上的重叠结合位点，在 E1 结合界面上。第一个系列的一个成员也与 E2 的转录激活域共结晶。对于这两个系列，都描述了结构活性研究，这导致活性提高了几百倍。每个系列中最好的化合物对 HPV11 E1-E2 相互作用的活性均在纳摩尔范围内，在细胞 DNA 复制测定中的 EC50 值约为 1μM。比较了这两个系列的结合模式，并从描述的工作中得出了关于发现蛋白-蛋白相互作用抑制剂的一些一般性结论。
