期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2015
卷号：112
期号：49
页码：15196-15201
DOI：10.1073/pnas.1513803112
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceA high-resolution structure was obtained for a drug candidate achieving pharmacological activity by inducing and stabilizing protein-protein interaction, a mechanism difficult to study in structural biology. We found that with poorly diffracting protein crystals, a protein stabilizing compound can improve crystal quality and enable the acquisition of a high-resolution structure. It also becomes apparent from this structure how improvements in pharmacologic potency can be achieved by improving protein-protein interaction stabilization and clear avenues for compound optimization are apparent from the data. The binding site observed in crystallography was biologically validated by mutational analysis, which also provides for the first time, to our knowledge, an understanding of a pathway by which viable, drug resistant virus variants may evolve against this drug class. The hepatitis B virus (HBV) core protein is essential for HBV replication and an important target for antiviral drug discovery. We report the first, to our knowledge, high-resolution crystal structure of an antiviral compound bound to the HBV core protein. The compound NVR-010-001-E2 can induce assembly of the HBV core wild-type and Y132A mutant proteins and thermostabilize the proteins with a Tm increase of more than 10 {degrees}C. NVR-010-001-E2 binds at the dimer-dimer interface of the core proteins, forms a new interaction surface promoting protein-protein interaction, induces protein assembly, and increases stability. The impact of naturally occurring core protein mutations on antiviral activity correlates with NVR-010-001-E2 binding interactions determined by crystallography. The crystal structure provides understanding of a drug efficacy mechanism related to the induction and stabilization of protein-protein interactions and enables structure-guided design to improve antiviral potency and drug-like properties.
